DNA sequences coding for the F18 fimbriae and AIDA adhesin are localised on the same plasmid in Escherichia coli isolates from piglets

The adhesin-involved-in-diffuse-adherence (AIDA) afimbrial adhesin is produced by human, but not by animal, Escherichia coli, with the exception of German porcine verotoxigenic Escherichia coli (VTEC) [Clin. Diagn. Lab. Immunol. 8 (2001) 143]. Presence and localisation of DNA sequences (aidA) coding for and production of an AIDA adhesin were investigated in a collection of Belgian VTEC and non-VTEC E. coli isolated from piglets at weaning time. The 174 isolates were also studied by colony hybridisation for the presence of DNA sequences coding for the Stx2e verocytotoxin and the F18 fimbrial adhesin (fed): 71 were Stx2+F18+AIDA+, 26 were F18+AIDA+, 12 were AIDA+, two were Stx2+AIDA+, and one was Stx2+ only. Fifty-four of the 58 (F18+)AIDA+ isolates tested positive in a western blotting assay with an immune serum raised against the AIDA protein. Hybridisation with the AIDA gene probe on plasmid DNA profiles identified a probe-positive plasmid band in the 10 AIDA+ and in 24 of the 25 F18+AIDA+ isolates studied. Moreover in F18+AIDA+ isolates, only one plasmid band hybridised with both F18 and AIDA probes. These results confirm the presence of aidA-related genes in not only VTEC, but also non-VTEC, isolates from piglets and the production of an antigenically AIDA-related protein by the majority of probe-positive E. coli. Moreover the plasmid DNA hybridisation results suggest a localisation on the same plasmid of the aidA- and fed-related DNA sequences.     

Mechanisms of resistance to fungicides in field strains of Botrytis cinerea

Field strains of Botrytis cinerea Pers ex Fr, the causal agent of grey mould diseases, were collected from French vineyards between 1993 and 2000. Several phenotypes have been characterized according to the inhibitory effects of fungicides towards germ-tube elongation and mycelial growth. Two types of benzimidazole-resistant strains (Ben R1 and Ben R2) could be detected; negative cross-resistance to phenylcarbamates (e.g. diethofencarb) was only found in Ben R1. Benzimidazole resistance was related to point mutations at codon 198 (Ben R1) or 200 (Ben R2) of the beta-tubulin gene. Most dicarboximide-resistant strains were also weakly resistant to aromatic hydrocarbon fungicides (e.g. dicloran) but remained sensitive to phenylpyrroles (e.g. fludioxonil). These resistant field strains (Imi R1) contained a single base pair mutation at position 365 in a two-component histidine kinase gene, probably involved in the fungal osmoregulation. Three anilinopyrimidine-resistant phenotypes have been identified. In the most resistant one (Ani R1), resistance was restricted to anilinopyrimidines, but no differences were observed in the amino-acid sequences of cystathionine beta-lyase (the potential target site of these fungicides) from Ani R1 or wild-type strains. In the two other phenotypes (Ani R2 and Ani R3), resistance extended to various other groups of fungicide, including dicarboximides, phenylpyrroles and sterol biosynthesis inhibitors. This multi-drug resistance was probably determined by over-production of ATP-binding cassette transporters. The hydroxyanilide fenhexamid is a novel botryticide whose primary target site is the 3-keto reductase involved in sterol C-4 demethylations. Apart from the multi-drug-resistant strain Ani R3, three other fenhexamid-resistant phenotypes have been recognized. For two of them (Hyd R1 and Hyd R2) fenhexamid-resistance seemed to result from P450-mediated detoxification. Reduced sensitivity of the target site could be the putative resistance mechanism operating in the third resistant phenotype (Hyd R3). Increased sensitivity to inhibitors of sterol 14 alpha-demethylase recorded in Hyd R1 strains was related to two amino-acid changes at positions 15 and 105 of this enzyme.     

Control and monitoring: Plum pox virus quarantine situation in France

Sharka disease caused by Plum pox virus (PPV) is present in several areas in France, and its total eradication from the territory is not considered feasible. Although sharka falls under mandatory control measures, the objective is, in the infected areas, to contain the disease in the orchards at sufficiently low levels so that the production remains economically viable. In parallel, official controls are carried out to guarantee that plants for planting, including seedlings, which are moved in trade are free from PPV.     

Effect of cultural practices on the development of arabica coffee berry disease,caused by <Emphasis Type="Italic">Colletotrichum kahawae</Emphasis>

In the high altitude regions of Africa, coffee berry disease (CBD), caused by Colletotrichum kahawae, is the main constraint for arabica coffee (Coffea arabica) production. However, certain agricultural practices can reduce losses caused by the disease and thereby promote optimum production. On small family farms in Cameroon, mixed cropping with fruit trees, intercropping with food crops and maintenance pruning of coffee trees are very widespread agricultural practices that can affect CBD epidemics. Consequently, an epidemiological study was conducted to assess how cultural practices affected the disease in an arabica coffee smallholding in Cameroon. The disease was monitored on a weekly basis over four successive years (2002–2005) on coffee trees in diverse cultural situations. Cultural practices likely to reduce losses due to CBD were identified. The infection rate was significantly lower on coffee trees grown intensively than on coffee trees grown in the traditional manner. Coffee trees located under the shade of fruit trees were significantly less attacked than those located in full sunlight. In addition, berries on the leafless parts of branches, near the main trunk of the coffee tree, were less infected than those on leafy sections. These results show that maintenance pruning, removal of mummified berries, and mixed cropping with shade plants are cultural practices which create environmental conditions that limit CBD development.     

Thermal optima of photosynthetic functions and thermostability of photochemistry in cork oak seedlings

Temperature effects on photosynthesis were studied in seedlings of evergreen Mediterranean cork oak (Quercus suber L.). Responses to changes in temperature and the temperature optima of maximal carboxylation rate (V(cmax)) and maximal light-driven electron flux (J(max)) were estimated from gas exchange measurements and a leaf-level photosynthesis model. The estimated temperature optima were approximately 34 and 33 degrees C for V(cmax) and J(max), respectively, which fall within the lower range of temperature optima previously observed in deciduous tree species. The thermostability of the photosynthetic apparatus was estimated according to the temperature at which basal chlorophyll a fluorescence begins to increase (T(c)). The T(c) was highly variable, increasing from 42 to 51 degrees C when ambient temperature rose from 10 to 40 degrees C, and increasing from 44 to 54 degrees C with decreasing soil water availability while net CO(2) assimilation rate dropped to almost zero. When a heat shock was imposed, an additional small increase in T(c) was observed in drought-stressed and control seedlings. Maximal T(c) values following heat shock were about 56 degrees C, which, to our knowledge, are the highest values that have been observed in tree species. In conclusion, the intrinsic temperature responses of cork oak did not differ from those of other species (similar T(c) under ambient temperature and water availability, and relatively low thermal optima for photosynthetic capacity in seedlings grown at cool temperatures). However, the large ability of cork oak to acclimate to drought and elevated temperature may be an important factor in the tolerance of this evergreen Mediterranean species to summer drought and high temperatures.     

Soil carbon dioxide efflux in pure and mixed stands of oak and beech

Total Soil Respiration (TSR) was measured in pure and mixed stands of oak and beech and was partitioned into two contributions using the forest floor removal technique: Mineral Soil Respiration (MSR) and Forest Floor Respiration (FFR). In addition, laboratory incubations of the forest floor and the Ah horizon were performed to evaluate the heterotrophic respiration and the DOC production of these horizons. The relationships between soil temperature and the various soil respiration contributions in the three stands were compared using Q ₁₀ functions. In situ, significant differences (α = 0,05) between stands were observed for the R ₁₀ parameter (respiration rate at 10 °C) of the TSR, MSR and FFR contributions, while only the temperature sensitivity (Q ₁₀) of TSR was significantly affected by stand composition. The effect of soil water content was only significant on MSR and followed different patterns according to stand composition. Under controlled conditions, the R ₁₀ of the forest floor and of the Ah horizon varied with stand composition and the Q ₁₀ of the forest floor decreased in the order: oak (2.27) > mixture (2.01) > beech (1.71).     

Field effect of P fertilization on N2 fixation rate of Ulex europaeus

European gorse (Ulex europaeus L.) N₂ fixation rate (%Ndfa) was studied in a maritime pine (Pinus pinaster Aït.) oligotrophic forest. Fertilization field trials were carried out on 5 sites with various inputs of phosphorus (0–240 kg P₂O₅·ha^?1). Seven to ten years after pine planting, gorse were sampled to evaluate the effect of P fertilization on gorse %Ndfa, determined using the ¹⁵N natural abundance method. One of the prerequisites of this method is the existence of a significant difference between the ¹⁵N/¹⁴N ratios in the atmospheric N reference and in the stand soil N references. This prerequisite was satisfied for 80 of 120 cases. The average %Ndfa was high (70 ± 3%) but with high local variability. No significant difference in %Ndfa was detected among P treatments. Nitrogen concentration of gorse was significantly higher in the highest dose treatments compared to the control.     

Anhydrous goat's milk fat: thermal and structural behaviors studied by coupled differential scanning calorimetry and X-ray diffraction. 2. Influence of cooling rate

Crystallization and melting properties of triacylglycerols (TGs) in anhydrous goat's milk fat (AGMF) are investigated by X-ray diffraction as a function of temperature (XRDT) coupled with high-sensitivity differential scanning calorimetry (DSC), using synchrotron radiation and Microcalix. The polymorphic behavior of AGMF was monitored by varying the cooling rates between 5 and 1 degrees C/min from 45 to -20 degrees C with their subsequent melting at 1 degrees C/min. Quenching of AGMF at -20 degrees C was also examined to determine the metastable polymorphic form of AGMF. At intermediate cooling rates, TGs in AGMF crystallize, from about 18 degrees C in two different lamellar structures with triple chain length 3Lalpha stacking of 72 A and a double chain length 2Lalpha stacking of 48 A, which are correlated to two overlapped exothermic peaks recorded by DSC. A reversible transition sub alpha <--> alpha was observed. Subsequent heating at 1 degrees C/min shows numerous structural rearrangements before final melting. At fast cooling of AGMF (5 degrees C/min), similar unstable crystalline varieties are formed while three endotherms are recorded. Several new unstable lamellar structures are observed after quenching. All of these data are compared to those previously reported at slow cooling (0.1 degrees C/min) showing a relative stability of the structures formed. In spite of general similitude, the thermal and structural behavior of the goat's milk is more complex than that of the cow's milk.     

Identification and quantification of seed carotenoids in selected wheat species

Selected primitive and modern wheat species were evaluated on the basis of their carotenoid composition and effects of the genotype and environment on lutein using spectrometry and liquid chromatography. Carotenoids in the wheat extracts were identified and confirmed on the basis of their UV/vis and mass spectra compared with those of authentic standards. The protonated molecule (M + 1)+ at m/z 569 was the predominant ion for zeaxanthin compared to the fragment ion at m/z 551 for lutein. A similar carotenoid profile was obtained for the wheat species investigated, but significant differences were observed in the concentration of carotenoids. Einkorn (Triticum monococcum) exhibited the highest level of all-trans-lutein, averaging 7.41 microg/g with small amounts of all-trans-zeaxanthin, cis-lutein isomers, and beta-carotene. Durum, Kamut, and Khorasan (Triticum turgidum) had intermediate levels of lutein (5.41-5.77 microg/g), while common bread or pastry wheat (Triticum aestivum) had the lowest content (2.01-2.11 microg/g). Lutein in einkorn appeared to be influenced significantly by environmental growing conditions.