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.     

Relationships between earthworm communities and burrow numbers under different land use systems

This study addresses the influence of three different land use systems (continuous maize, pasture/maize rotation, permanent pasture) on the relationships between earthworm populations and the number of earthworm burrows quantified in a soil profile. Quantified burrows were limited to those observable by the naked eye (i.e. >2 mm in diameter) and enumerated earthworms were limited to those which could have created the observable burrows (i.e. >0.3 g).The results were combined with data from the literature coming from different geographical regions. This study showed that earthworm abundance decreased with the increasing land management intensity (maize crop vs. pasture), while the number of burrows could be higher or similar under maize compared to pasture. Under maize, despite lower earthworm abundance and the annual destruction of the burrows by tillage, the number of burrows was almost as high as under pasture. This absence of a relationship between burrow numbers and earthworm abundance was observed in the soil profile and for each layer of the profile for each land use system. Furthermore, the burrow number/m^?2 per earthworm strongly varied depending on land use and was far higher under maize when compared to pasture (74 vs. 7). Therefore, a power-law type relationship was clearly established between burrow number/m^?2 per earthworm and earthworm abundance. This power type relationship was also observed when including data from the literature although it followed a different mathematical model. These results were explained by (i) increased earthworm burrowing activity (i.e. an increase in the number of burrows produced by each earthworm) under maize, and (ii) the dynamics of burrow number under pasture (i.e. decreased burrowing activity and burrow destruction process); both results of food accessibility combined with inter-individual competition. The results of the study suggest that farmers should not use the number of pores as an indicator of earthworm abundance, but as an indicator of earthworm activity, which could be integrated in an indicator of soil quality.     

Analytical and sensorial characterization of the aroma of wines produced with sour rotten grapes using GC-O and GC-MS: identification of key aroma compounds

In the present work, the aroma profiles of wines elaborated from sound and sour rot-infected grapes as raw material have been studied by sensory analysis, gas chromatography-olfactometry (GC-O), and gas chromatography-mass spectrometry (GC-MS), with the aim of determining the odor volatiles most likely associated with this disease. The effect of sour rot was tested in monovarietal wines produced with the Portuguese red grape variety Trincadeira and in blends of Cabernet Sauvignon and sour rotten Trincadeira grapes. Wines produced from damaged berries exhibited clear honey-like notes not evoked by healthy samples. Ethyl phenylacetate (EPhA) and phenylacetic acid (PAA), both exhibiting sweet honey-like aromas, emerged as key aroma compounds of sour rotten wines. Their levels were 1 order of magnitude above those found in controls and reached 304 and 1668 μg L(-1) of EPhA and PAA, respectively, well above the corresponding odor thresholds. Levels of γ-nonalactone also increased by a factor 3 in sour rot samples. Results also suggest that sour rot exerts a great effect on the secondary metabolism of yeast, decreasing the levels of volatiles related to fatty acids and amino acid synthesis. The highest levels of γ-decalactone of up to 405 μg L(-1) were also found in all of the samples, suggesting that this could be a relevant aroma compound in Trincadeira wine aroma.     

Adapting ecotoxicological tests based on earthworm behavior to assess the potential effectiveness of forest soil liming

Two laboratory experiments were carried out to determine the potential effects of lime application on the behavior of earthworms inoculated in an acidified forest soil from Vosges Mountains. Several field soils were studied: (i) a non-limed soil that had received decades of atmospheric acid depositions $({\text{pH}}_{{\text{H}}_2\text{O}}=3.9)$, (ii) an in situ limed soil that had been limed 6 years before at 2.5 t ha^?1 $({\text{pH}}_{{\text{H}}_2\text{O}}=4.2)$ and (iii) in vitro limed soils composed of non-limed soil mixed with several lime amounts in the laboratory corresponding to field rates of 1, 2.5, 5, 10 and 20 t ha^?1. Firstly, we adapted the earthworm avoidance test (ISO, 2006) by using Eisenia fetida as the model organism and Lumbricus castaneus, a local species, to determine earthworm preference between non-limed and limed soils. Secondly, a cast production (CP) study was performed according to Capowiez et al. (2009) with Lumbricus terrestris, in seven different treatments (non-limed soil, in situ limed and five in vitro limed soils with pH from 5 to 6.3). Both species avoided the non-limed soil in favour of in situ and in vitro limed soils for both species. Results of the CP bioassay showed that the bioturbation behavior of L. terrestris significantly increased with the increase of soil pH, following a dose–response curve. However, in the long term (>6 years), the application of lime was insufficient to significantly improve soil conditions for enhancing earthworm activity. In conclusion, we may recommend this kind of earthworm laboratory tests with field soils to assess any changes in soil quality over time due to liming application.     

Post-fire attractiveness of maritime pines (<Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait.) to xylophagous insects

The spectrum and dynamics of xylophagous insects infesting Pinus pinaster after a fire event were studied from November 2007 to December 2008 at Sierra de Outes (Galicia, NW Spain). Insects were collected within and outside the fire area. Relationships among species status, species abundance, tree parameters and tree injury were assessed. Mortality of injured trees was also recorded. Insect colonization built up consistently during the first year after the fire, although the pests did not colonize unburned trees in nearby areas. Thirteen insect species from six xylophagous families were collected. Tomicus spp., Buprestis novenmaculata, Anobium punctatum and Pissodes castaneus were the first species detected. Tomicus spp. (29%) and Ips sexdentatus (23%) were the dominant taxa. Bole char height and soil burn severity were positively correlated with insect presence. Trees with short stem diameter and thinner bark were also preferred. B. novenmaculata presence was positively correlated with crown scorch. Loss of cambium hydration was negatively related to the occurrence of pests, likely because sapwood desiccation and loss of nutrient impede brood development. About 33.3% of the fire-scorched pines died. The results recommend the removal of injured and dying pines after fire, to avoid the rise of pest population threatening recovering trees in the burned areas.     

Performance of four <Emphasis Type="Italic">Trichogramma</Emphasis> species (Hymenoptera: Trichogrammatidae) as biocontrol agents of <Emphasis Type="Italic">Heliothis virescens</Emphasis> (Lepidoptera: Noctuidae) under various temperature regimes

Trichogramma spp. are major parasitoids of lepidopteran pest eggs, but there is large variation in efficacy toward a given pest among the numerous described Trichogramma species. It is important to select the Trichogramma species that most effectively parasitize and develop in target pest eggs for biological control. In this context, Trichogramma pretiosum, T. exiguum, T. atopovirilia and T. acacioi were studied in Heliothis virescens eggs under different thermal conditions. The parasitoids were reared at constant temperatures of 20, 25 and 30°C and tested at these respective temperatures, while parasitoids reared at 25°C were also tested at 20 and 30°C, for a total of 20 species–temperature combinations. About 30 H. virescens eggs were offered to the parasitoids for 24 h. Among the four species, parasitism rate by T. atopovirilia was highest at all temperature conditions, whereas T. acacioi had the lowest rates of parasitism at 25°C and 25/30°C. Parasitism ranged from 13.8 to 43.8% among all species–temperature combinations. Viability (emerged parasitoids) ranged from 80.8 to 98.4%, and was deemed satisfactory. The emergence rates of T. exiguum and T. acacioi were affected by temperature. Temperature also affected the sex ratio of T. exiguum at 25/30°C, whereas T. pretiosum and T. acacioi produced females predominantly independent of temperature. Overall, the parasitoid T. atopovirilia was the most efficient in parasitizing H. virescens eggs, though the levels of parasitism obtained might not ensure its successful use in biological control programs. The temperature-related differences in biological traits observed in the four Trichogramma species tested hint at the importance of making careful choices regarding climatic conditions where the parasitoid is going to be used when selecting a species for biological control programs.     

Larval performances and life cycle completion of the Siberian moth, <Emphasis Type="Italic">Dendrolimus sibiricus</Emphasis> (Lepidoptera: Lasiocampidae), on potential host plants in Europe: a laboratory study on potted trees

The Siberian moth, Dendrolimus sibiricus, Tschtv. is the most harmful defoliator of coniferous forests in North Asia. The pest has already spread over the Urals and continues moving westwards. Recently, it has been recommended for quarantine in member countries by European and Mediterranean Plant Protection Organization (EPPO). The performances of the pest on coniferous species planted in Europe were assessed on a range of potted trees corresponding to the spectrum of economically important conifers in the EU: European larch Larix decidua, Norway spruce Picea abies, Scots pine Pinus sylvestris, European black pine Pinus nigra, and the North American species: Douglas fir Pseudotsuga menziesii and grand fir Abies grandis. Larvae showed a potential to survive and complete the development on all these host tree species. Favorable hosts were grand fir, European larch, and Douglas fir that allowed higher survival, better larval development, and as a result, yielded heavier pupae and adult moths with higher longevity. Black pine was a poor host but, however, could still support larval and pupal development. Norway spruce and Scots pine had an intermediate behavior. If accidentally introduced to Europe, the Siberian moth may become especially damaging in forest stands predominated by European larch and by the North American firs. Norway spruce and especially the two-needle pines will be less prone to intensive defoliation by this species. The fact that the pest may damage the range of economically important coniferous species should be taken into account in the pest risk assessment for Europe and also for North America where the Siberian moth occurrence is considered likely.     

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.