Adaptation to fungicides in Monilinia fructicola isolates with different fungicide resistance phenotypes

The ability to develop fungicide resistance was assessed in Monilinia fructicola isolates with different fungicide sensitivity phenotypes by adapting mycelium and conidia to increasing concentrations of selective fungicides and UV mutagenesis. Results showed that adaptation to Quinone outside inhibitor (QoI) fungicide azoxystrobin and sterol demethylation inhibitor (DMI) fungicide propiconazole was more effective in conidial-transfer experiments compared to mycelial-transfer experiments. DMI-resistant (DMI-R) isolates adapted to significantly higher doses of azoxystrobin in both, mycelial- and conidial-transfer experiments compared to benzimidazole-resistant (BZI-R) and sensitive (S) isolates. Adaptation to propiconazole in conidial-transfer experiments was accelerated in BZI-R isolates when a stable, nonlethal dose of 50 microg/ml thiophanate-methyl was added to the selection medium. One of two azoxystrobin-resistant mutants from DMI-R isolates did not show any fitness penalties; the other isolate expired before further tests could be carried out. The viable mutant caused larger lesions on detached peach fruit sprayed with azoxystrobin compared to the parental isolate. The azoxystrobin sensitivity of the viable mutant returned to baseline levels after the mutant was transferred to unamended medium. However, azoxystrobin resistance recovered quicker in the mutant compared to the corresponding parental isolate after renewed subculturing on medium amended with 0.2 and 1 microg/ml azoxystrobin; only the mutant but not the parental isolate was able to adapt to 5 microg/ml azoxystrobin. In UV mutagenesis experiments, the DMI-R isolates produced significantly more mutants compared to S isolates. All of the UV-induced mutants showed stable fungicide resistance with little fitness penalty. This study indicates the potential for QoI fungicide resistance development in M. fructicola in the absence of a mutagen and provides evidence for increased mutability and predisposition to accelerated adaptation to azoxystrobin in M. fructicola isolates resistant to DMI fungicides.     

Seasonal variation of red clover (Trifolium pratense L., Fabaceae) isoflavones and estrogenic activity

Red clover (Trifolium pratense L., Fabaceae) dietary supplements are currently used to treat menopausal symptoms because of their high content of the mildly estrogenic isoflavones daidzein, genistein, formononetin, and biochanin A. These compounds are estrogenic in vitro and in vivo, but little information exists on the best time to harvest red clover fields to maximize content of the isoflavones and thus make an optimal product. Samples of cultivated red clover above-ground parts and flower heads were collected in parallel over one growing season in northeastern Illinois. Generally, autohydrolytic extracts of above-ground parts contained more isoflavones and had more estrogenic activity in Ishikawa endometrial cells as compared with extracts of flower heads. Daidzein and genistein contents peaked around June to July, while formononetin and biochanin A contents peaked in early September. Flower head and total above-ground parts extracts exhibited differential estrogenic activity in an Ishikawa (endometrial) cell-based alkaline phosphatase induction assay, whereas nondifferential activity was observed for most extracts tested in an MCF-7 (breast) cell proliferation assay when tested at the same final concentrations. Ishikawa assay results could be mapped onto the extracts' content of individual isoflavones, but MCF-7 results did not show such a pattern. These results suggest that significant metabolism of isoflavones may occur in MCF-7 cells but not in Ishikawa cells; therefore, caution is advised in the choice of bioassay used for the biological standardization of botanical dietary supplements.     

Anticipatory reproduction and population growth in seed predators

Mast seeding, the intermittent, synchronous production of large seed crops by a population of plants, is a well-known example of resource pulses that create lagged responses in successive trophic levels of ecological communities. These lags arise because seed predators are thought capable of increasing reproduction and population size only after the resource pulse is available for consumption. The resulting satiation of predators is a widely cited explanation for the evolution of masting. Our study shows that both American and Eurasian tree squirrels anticipate resource pulses and increase reproductive output before a masting event, thereby increasing population size in synchrony with the resource pulse and eliminating the population lag thought to be universal in resource pulse systems.     

Antioxidant and anti-inflammatory capacity of bioaccessible compounds from wheat fractions after gastrointestinal digestion

Wholegrain consumption is associated with several health benefits, in contrast to the consumption of refined grains. This can partly be related to the antioxidant compounds in the outer parts of the grain kernel. The bioaccessibility of these antioxidant compounds from the wholegrain matrix during gastrointestinal digestion is crucial for their absorption and bioavailability. In the current study, the bioaccessible compounds from aleurone, bran and flour were obtained from a dynamic in vitro model of the upper gastrointestinal tract. They were collected at 1 h time intervals to assess their antioxidant capacity (TEAC assay) and also their anti-inflammatory effect (TNF-α reduction in U937 macrophages stimulated with LPS). The bioaccessible compounds from aleurone had the highest antioxidant capacity and provided a prolonged anti-inflammatory effect, shown by the TNF-α reduction of a relatively late time-interval (3–4 h after start of digestion). The contribution of ferulic acid to those effects was minor due to its low bioaccessibility. Aleurone seems a promising wheat fraction for cereal products with a healthy added value.     

Population genetic structure of common bottlenose dolphins (Tursiops truncatus) in the Adriatic Sea and contiguous regions: implications for international conservation

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An intron in the cytochrome b gene of Monilinia fructicola mitigates the risk of resistance development to QoI fungicides

BACKGROUND: The cytochrome b (Cyt b) gene is a key genetic determinant for quinone outside inhibitor (QoI) fungicide resistance in plant pathogenic fungi. A mutation at amino acid position G143 can cause qualitative resistance unless it is part of the recognition site for a self‐splicing intron. The objective of this study was to clone and sequence the Cyt b gene from Monilinia fructicola (Wint.) Honey, the causal agent of brown rot of stone fruits, and to assess the risk for the development of a mutation at position 143. RESULTS: The Cyt b gene of M. fructicola was 11 927 bp in size and contained seven introns located at cDNA positions (5′–3′) 204, 395, 430, 491, 507, 780 and 812 with sizes of 1592, 1318, 1166, 1252, 1065, 2131 and 2227 bp respectively. Sequence analysis revealed that the above‐mentioned 1166 bp intron, a self‐splicing group I intron, was located just downstream of the G143 codon. The Cyt b gene region covering the G143 location and the adjacent 1166 bp intron was PCR amplified and sequenced from Chinese and US isolates, indicating that the intron may be omnipresent in M. fructicola. CONCLUSION: This is the first complete Cyt b gene sequence published for M. fructicola or any other Monilinia species, forming the basis for molecular analysis of QoI fungicide resistance. Sequence analysis revealed that the G143A mutation responsible for high levels of QoI fungicide resistance in many plant pathogenic fungi may not develop in M. fructicola unless genotypes emerge that lack the 1166 bp intron. Copyright © 2010 Society of Chemical Industry     

Application of the membrane interphase probe (MIP): an evaluation

Background, aim, and scope

The membrane interphase probe (MIP?) from Geoprobe Systems® has frequently been applied in different countries for the characterization of soil contaminated with volatile organic carbons (VOCs). Experience shows that misinterpretation of the collected data is common. This is mainly due to the lack of understanding and knowledge related to the detectors used, their detection limits, and the sensitivity of the MIP system. It has been noticed that the sensitivity of the system given by the producer and by different users are rather optimistic, e.g., the values given are lower (= better) than those actually experienced in the field. A need for a better understanding of the MIP system sensitivity, combined with a more scientifically based interpretation of the collected data, exists.

Materials and method

Both laboratory tests (using solutions) as well as field measurements were carried out using different detector configurations to allow a better interpretation of the detector signals/system sensitivity and to collect qualitative information. These configurations were: (1) detectors stand alone; (2) the use of a 2-ml sample loop, and (3) a purge and trap system. The configurations (2) and (3) are used in combination with a capillary column to carry out on-site qualitative and semiquantitative analyses.

Results and discussion

With respect to the configuration of “detectors stand alone,” detection limits for toluene (in aqueous solutions) range between 4 ppm (flame ionization detector—FID) and 10 ppm [photo ionization detector (PID)]. For chlorinated aliphatic hydrocarbons (CAHs), observed limits are 10 ppm (FID), 4–50 ppm (PID), and 3–10 ppm [dry electrolytic conductivity detector (DELCD)]. When using the 2-ml sample loop, relatively high concentrations have to be initially present in the soil. Observed detection limits for mono-aromatic hydrocarbons are 5–100 ppm; for CAHs, 1–50 ppm; for alkanes, 250–400 ppm; and for MTBE, 25 ppm. The application of purge and trap results in a better resolution and the detection of lower concentrations. Consequently, a better identification of the pollution with depth is possible. In this case, the detection limits are a function of the concentrations and the flushing time. In relation to the qualitative analyses, it was found that the configuration of the MIP-system with the built-in capillary column and the 2-ml sample loop or the purge and trap preconcentrator, respectively, are useful to carry out on-site analyses, thus allowing a better identification of the pollution in a vertical profile.

Conclusions

The measurements carried out using the MIP with detectors stand-alone or in combination with a loop or trap, or connected to a column, confirm that analysis is indeed very useful to characterize VOC source zones when knowing and understanding its performance. This relates mainly to the detection limits of the MIP system. For a selection of parameters, such limits have been obtained. These values seem to be more realistic than those found in the few references where numbers are given. For the qualitative measurements, it can be concluded that a better resolution is obtained, and pollutants present in lower concentrations will be detected when using the purge and trap. It is advised to determine the optimal flushing time and the detection limit of the expected pollutants in advance.

Recommendations and perspectives

This study indicates that there is still a need for further measurements and discussion between users. Finally, additional data should result in a better interpretation of the collected field data.     

Detection and identification of beta-lactam residues in milk using a hybrid biosensor

A novel application of a hybrid biosensor is here employed as an analytical method for the detection and presumptive identification of beta-lactam residues in milk. The method is based on measurements of carbon dioxide (CO2), the production of which is related to the microbial growth of the test microorganism Bacillus stearothermophilus var. calidolactis. The presence of beta-lactams in milk inhibits microbial growth and, consequently, the CO2 production rate. The analysis is based on the variation of CO2 between a milk sample spiked with beta-lactams and a twin milk sample containing beta-lactams plus a broad spectrum beta-lactamase, using an electrochemical device of biosensor. A blank milk sample is included as control. The result is obtained starting from the first 120 min. Moreover, the ability to recognize all of the beta-lactams speeds the total time of analysis when chemical identification and quantification are required. The analytical method appears to be adequate for milk control for qualitative screening purposes, complying with the requirements stated in Decision 2002/657/EC.     

Podzolization as a deferralitization process: dynamics and chemistry of ground and surface waters in an Acrisol – Podzol sequence of the upper Amazon Basin

Hydrochemical processes involved in the development of hydromorphic Podzols are a major concern for the upper Amazon Basin because of the extent of the areas affected by such processes and the large amounts of organic carbon and associated metals exported to the rivers. The dynamics and chemical composition of ground and surface waters were studied along an Acrisol‐Podzol sequence lying in an open depression of a plateau. Water levels were monitored along the sequence over a period of 2 years by means of piezometers. Water was sampled in zero‐tension lysimeters for groundwater and for surface water in the drainage network of the depression. The pH and concentrations of organic carbon and major elements (Si, Fe and Al) were determined. The contrasted changes reported for concentrations of Si, organic carbon and metals (Fe, Al) mainly reflect the dynamics of the groundwater and the weathering conditions that prevail in the soils. Iron is released by the reductive dissolution of Fe oxides, mostly in the Bg horizons of the upslope Acrisols. It moves laterally under the control of hydraulic gradients and migrates through the iron‐depleted Podzols where it is exported to the river network. Aluminium is released from the dissolution of Al‐bearing minerals (gibbsite and kaolinite) at the margin of the podzolic area but is immobilized as organo‐Al complexes in spodic horizons. In downslope positions, the quick recharge of the groundwater and large release of organic compounds lead to acidification and a loss of metals (mainly Al), previously stored in the Podzols.