Biosynthesis of Depsipeptide Mycotoxins in Fusarium

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.     

The Effect of Inoculation Treatment and Long-term Application of Moisture on Fusarium Head Blight Symptoms and Deoxynivalenol Contamination in Wheat Grains

Fusarium head blight (FHB) is an important disease of wheat, which can result in the contamination of grains with mycotoxins such as deoxynivalenol (DON). Artificial inoculation of flowering ears with conidial suspensions is widely used to study FHB diseases. Our goal was to compare four inoculation treatments in which a conidial suspension was sprayed on flowering ears and to study the effect of the application of moisture during kernel setting and filling with a mist-irrigation system. Ten wheat genotypes were inoculated with a DON-producing Fusarium culmorum strain. Inoculation treatments varied in time of application of the inoculum (morning or evening) and in the method of controlling humidity during inoculation (bagging or mist irrigation). A wet season was simulated with a mist-irrigation system, keeping the crop canopy wet for at least 26 days after flowering. The severity of FHB symptoms (area under disease progress curve (AUDPC)), yield loss and DON contamination in the grains were determined. AUDPC data obtained with the different inoculation treatments were highly correlated (r=0.85–0.95). Mist irrigation after inoculation resulted in a higher mean disease severity, but in a overall lower toxin contamination as compared to the non-irrigated treatments. Genotypic differences in DON accumulation were present: for one wheat line toxin contamination significantly increased when irrigated, while two genotypes accumulated significantly less toxin. The closest relationships (r=0.73–0.89) between the visual symptoms and the DON content were obtained under moderate mean infection pressure. This relation between visual symptoms and the DON content deteriorated at higher infection levels.     

Evaluation of a combined transcutaneous carbon dioxide pressure and pulse oximetry sensor in adult sheep and dogs

OBJECTIVE: To evaluate a combined transcutaneous carbon dioxide pressure (tcPCO(2)) and pulse oximetry sensor in sheep and dogs. ANIMALS: 13 adult sheep and 11 adult dogs. PROCEDURES: During inhalation anesthesia, for the first 10 minutes following sensor placement, arterial blood gas was analyzed and tcPCO(2) was recorded every 2 minutes. Subsequently, the animals were hyper-, normo-, and hypoventilated. The simultaneously obtained tcPCO(2) and PaCO(2) values were analyzed by use of Bland-Altman statistical analysis. RESULTS: Mean +/- SD overall difference between tcPCO(2) and PaCO(2) 10 minutes after sensor application was 13.3 +/- 8.4 mm Hg in sheep and 8.9 +/- 12 mm Hg in dogs. During hyper-, normo-, and hypoventilation, mean difference (bias) and precision (limits of agreement [bias +/- 2 SD]) between tcPCO(2) and PaCO(2) values were 13.2 +/- 10.4 mm Hg (limits of agreement, -7.1 and 33.5 mm Hg) in sheep and 10.6 +/- 10.5 mm Hg (limits of agreement, -9.9 and 31.2 mm Hg) in dogs, respectively. Changes in PaCO(2) induced by different ventilation settings were detected by the tcPCO(2) sensor with a lag (response) time of 4.9 +/- 3.5 minutes for sheep and 6.2 +/- 3.6 minutes for dogs. CONCLUSIONS AND CLINICAL RELEVANCE: The tcPCO(2) sensor overestimated PaCO(2) in sheep and dogs and followed changes in PaCO(2) with a considerable lag time. The tcPCO(2) sensor might be useful for noninvasive monitoring of changes but cannot be used as a surrogate measure for PaCO(2).     

Stability and Biological Activity of Cyfluthrin Isomers

A GC-MS method capable of completely separating the four pairs of diastereoisomers of cyfluthrin is presented and the method used to show that isomerisation of the cyfluthrin enantiomers occurs in methanol. This methanol-induced isomerisation could also be demonstrated by bioassays using water fleas. The biological activities of the various cyfluthrin isomers contained in the commercial products cyfluthrin and beta-cyfluthrin were assayed using several strains of lepidopteran larvae including Plutella xylostella, Heliothis virescens and Spodoptera frugiperda. With the susceptible strains, the efficiencies of the isomers mixtures of cyfluthrin and beta-cyfluthrin were shown to obey the rules of additivity. However, in tests with a resistant strain of P. xylostella originating from Thailand, the ‘inactive’ isomer III acted synergistically with the active isomer IV. Resistance factors in strains of H. virescens and P. xylostella were found to be higher with cis than with trans isomers. This probably contributes to the superior action of cyfluthrin and beta-cyfluthrin against various pests of agricultural importance since the commercial products contain a high content of trans isomers (‘high trans pyrethroids’).     

Modelling compaction of agricultural subsoils by tracked heavy construction machinery under various moisture conditions in Switzerland

In recent years, agricultural land in Switzerland has been increasingly used as temporary access ways for heavy machinery in road and pipeline construction operations. The Swiss soil protection law requires that measures are taken to prevent soil compaction in such operations, but gives no criteria to determine tolerable loads. We studied the compaction sensitivity of a loess soil (Haplic Luvisol) at different soil moisture conditions in a field traffic experiment and by a numerical model on the computer using finite element analysis. Two plots, one wetted by sprinkling and one left dry (no sprinkling), were traversed by heavy caterpillar vehicles during construction of a large overland gas pipeline. Compaction effects were determined by comparing precompression stresses of samples taken from trafficked and non-trafficked soil. A finite element model with a constitutive relation, based on the concept of critical state soil mechanics, was used to interpret the outcome of the field trials.

We found significantly higher precompression stresses in the trafficked (median 97 kPa) compared with the non-trafficked (median 41 kPa) topsoil of the wet plot. No effect was evident in the topsoil of the dry plot as well as in the subsoils of the wet and the dry plot. The observed compaction effects were in agreement with the model predictions if the soil was assumed to be partially drained, but disagreed for the wet subsoil if fully drained conditions were assumed. Agreement between model and experimental results also required that the moisture dependence of the precompression stress was taken into account.     

Microbial use of organic amendments in saline soils monitored by changes in the C/C ratio

An incubation experiment was carried out to investigate whether salinity at high pH has negative effects on microbial substrate use, i.e. the mineralization of the amendment to CO₂ and inorganic N and the incorporation of amendment C into microbial biomass C. In order to exploit natural differences in the ¹³C/¹²C ratio, substrate from two C₄ plants, i.e. highly decomposed and N-rich sugarcane filter cake and less decomposed N-poor maize leaf straw, were added to two alkaline Pakistani soils differing in salinity, which had previously been cultivated with C₃ plants. In soil 1, the additional CO₂ evolution was equivalent to 65% of the added amount in the maize straw treatment and to 35% in the filter cake treatment. In the more saline soil 2, the respective figures were 56% and 32%. The maize straw amendment led to an identical immobilization of approximately 48 μg N g⁻¹ soil over the 56-day incubation in both soils compared with the control soils. In the filter cake treatment, the amount of inorganic N immobilized was 8.5 μg N g⁻¹ higher in soil 1 than in soil 2 compared with the control soils. In the control treatment, the content of microbial biomass C₃-C in soil 1 was twice that in soil 2 throughout the incubation. This fraction declined by about 30% during the incubation in both soils. The two amendments replaced initially similar absolute amounts of the autochthonous microbial biomass C, i.e. 50% of the original microbial biomass C in soil 1 and almost 90% in soil 2. The highest contents of microbial biomass C₄-C were equivalent to 7% (filter cake) and 11% (maize straw) of the added C. In soil 2, the corresponding values were 14% lower. Increasing salinity had no direct negative effects on microbial substrate use in the present two soils. Consequently, the differences in soil microbial biomass contents are most likely caused indirectly by salinity-induced reduction in plant growth rather than directly by negative effects of salinity on soil microorganisms.     

Biochemical and toxicological differences in the modes of action of the benzoylureas

The two insecticidal benzoylurea compounds, diflubenzuron and chlorfluazuron, show large differences in their toxicity against the larvae of insects like the tobacco budworm, Heliothis virescens, or the Egyptian cotton leafworm, Spodoptera littoralis, chlorfluazuron being about 100 times more toxic. This difference is due mainly to a much faster metabolism of diflubenzuron. Its half-life within the larvae is about 5 h, compared to about 50 h for chlorfluazuron. Chlorfluazuron is also the much better ovicide of the two, following injection of the compounds into the females of H. virescens. Again the difference in the rate of metabolism is the main cause. The rate of excretion of the parent benzoylureas is relatively low, but their metabolites are excreted very quickly.