Dominant rhizosphere bacteria as a source for antifungal agents

A search for micro-organisms associated in high numbers with roots or leaves of corn, grapevine, chicory, soybean, sunflower, barley and sugarbeet yielded a collection of over 10 000 bacterial strains. Within this collection, antifungal strains have been selected, using direct or indirect in-vitro assays against one target fungus per crop. The target fungi were selected based on their agronomic relevance as pests. Isolates with antifungal activity were tested for their spectrum of activity against a set of phytopathogenic fungi. In 15 bacterial strains with broad-spectrum activity, belonging to the species Erwinia herbicola. Serratia plymuthica, Pseudomonas fluorescens, P. aureofaciens, P. cepacia and Bacillus subtilis, the active compounds have been identified. Bacterial strains belonging to the same taxon, but isolated from different ecological niches and/or different places in Europe, produce identical compounds. In two bacterial taxa (Erwinia and Serratia) this was reflected in a genetic conservation of the regions coding for antifungal activity. In both cases, the biosynthetic pathways proved to be genetically complex. All of the compounds identified in this study have been described previously in bacterial strains isolated from analogous or completely different ecosystems.     

Rapid, multiwell colorimetric assay for measuring neutrophil chemoattractant activity in bronchoalveolar lavage fluid of horses with recurrent airway obstruction.

The criteria used to diagnose recurrent airway obstruction (RAO) in affected horses include demonstration of reversible lower airway obstruction and greater than 25% neutrophils in bronchoalveolar lavage fluid (BALF). Additional objective laboratory tests are needed to improve diagnostic accuracy and to monitor response to treatment. The goal of this study was to determine if neutrophil chemoattractant activity of BALF could be measured by using a previously described, rapid, multiwell colorimetric assay for chemotaxis. In this assay, neutrophils that have migrated through a membrane filter are collected into the bottom well of a disposable chemotaxis-cell migration chamber. The number of viable cells collected in the bottom well is quantified by measurement of the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenol tetrazolium bromide (MTT), which is reduced by dehydrogenase in mitochondria of live cells. The number of migrating cells corresponds to the amount of MTT reduced, which is measured with an enzyme-linked immunosorbent assay plate reader. Fourteen adult horses were enrolled in this study, 7 of which had owner histories consistent with RAO. Each horse was sedated, a bronchoalveolar lavage tube was passed, and saline was infused and immediately aspirated. An aliquot of BALF was used for differential cell count, and BALF supernatant was harvested to assess neutrophil chemoattractant activity. Normal control horses and RAO-affected horses were distinguished according to clinical signs and percent neutrophils in BALF. Neutrophil chemoattractant activity of BALF was significantly greater in RAO-affected horses (P = 0.001) compared with control horses. This assay may be useful in future studies for monitoring response to therapy in RAOaffected horses.     

Characterisation of bound residues of [3H]triforine in barley grain grown in the field

Barley was grown in an experimental field and at growth stage J (during the stem extension stage when the second node of the stem was formed and the next-to-last leaf was just visible), the aerial part of the plants was sprayed with an aqueous emulsion of a mixture of triforine and [³H]triforine (uniformly labelled in the piperazine ring) at the normal rate of 250 g triforine ha^?1. The barley was harvested when ripe, and the grain was analysed separately. Extraction of the grain with methanol left methanol-insoluble solids containing an amount of radioactivity (the bound residue) which represented 75% of the total radioactivity incorporated into the grain. Methanol acidified with hydrochloric acid extracted a further 7% of the triforine-derived bound residues in the form of radioactive iminodiacetic acid (1.1%), glycine (3.3 %), serine (0.9%), ethanolamine (0.2%) and unidentified compounds (1.5 %); in the grain, these compounds or their precursors had thus been complexed to grain constituents. Aqueous 0.03M sodium hydroxide extracted a further 27% of the total tritium which had been incorporated by means of α chemical bonds into the protein fraction; acid hydrolysis of the proteins yielded radioactive glycine (9.2%), serine (3.9%) and unidentified compounds (13.9%) which could have been a mixture of a large number of other amino-acids. The plant solids (which contained 41% of the total tritium) left after the alkaline aqueous extractions, were processed and separated into tritiated cellulose (4%) and starch (37%) fractions. The starch was hydrolysed and the resulting glucose was converted into the osazone (34%). After being recrystallised several times, the osazone contained a constant specific radioactivity, indicating that [³H]glucose was present. This glucose may be considered as having a carbon skeleton mainly originating biochemically from some metabolites of [³H]triforine; this tritiated glucose may also be considered as indirect evidence of the biochemical oxidation, by transamination, of the metabolites of [³H]triforine; in barley grain, the tritiated glucose (or at least a part of it) was anabolised into proteins. No piperazine was observed in the bound residues in the grain.     

Flow cytometry: An aid in monitoring lymphocyte transformation in whole blood cultures

A combination of whole blood cultures, hypotonic fluorescent staining of nucleated cells, and flow cytometry resulted in a greatly improved technique for measuring lymphocyte blastogenesis. Sample preparation was reduced to two steps with no centrifugation. The technique was successfully used to analyze blood cultures derived from experimentally and naturally infected cows. The technique was also shown to be ideally suited for analyzing lymphocytes cultured in whole blood cultures and (1) avoids the problems of tritiated thymidine analysis and (2) provides a direct analysis of the number of cells responding to antigenic stimulation.     

Osseous sequestration in cattle: 110 cases (1987-1997)

OBJECTIVE: To determine risk factors for development of sequestra in cattle and identify factors associated with a successful outcome. DESIGN: Retrospective study. ANIMALS: 110 cattle. PROCEDURE: Medical records of cattle treated at veterinary teaching hospitals in North America were reviewed. To determine risk factors for osseous sequestration, breed, age, and sex of cattle with osseous sequestration were compared with breed, age, and sex of all other cattle admitted during the study period. RESULTS: 110 cattle were included in the study. Three had 2 sequestra; thus, 113 lesions were identified. Most sequestra were associated with the bones of the extremities, most commonly the third metacarpal or third metatarsal bone. Ninety-two animals were treated surgically (i.e., sequestrectomy), 7 were treated medically, 3 were initially treated medically and were then treated surgically, and 8 were not treated. Follow-up information was available for 65 animals treated surgically and 6 animals treated medically. Fifty-one (78%) animals treated surgically and 5 animals treated medically had a successful outcome. Cattle that were 6 months to 2 years old had a significantly increased risk of developing a sequestrum, compared with cattle < 6 months old. Cattle in which sequestrectomy was performed with the aid of local anesthesia were significantly more likely to undergo 2 or more surgical procedures than were cattle in which sequestrectomy was performed with the aid of general anesthesia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that sequestrectomy will result in a successful outcome for most cattle with osseous sequestration.     

Role of the ejaculatory bulb in biosynthesis of the male pheromone cis‐vaccenyl acetate in Drosophila melanogaster

In Drosophila melanogaster, the male ejaculatory bulb is the site of synthesis of a male‐specific pheromone, cis‐vaccenyl acetate, which functions as both an attractant and an anti‐aphrodisiac. This long monounsaturated acetate is structurally similar to a number of shorter gland‐synthesized moth pheromones. The cell monolayer that forms the Drosophila male ejaculatory bulb wall is responsible for the production and secretion of cis‐vaccenyl acetate into the seminal fluid. When dissected bulbs were incubated with sodium [14‐C]‐acetate (or deuterated acetate), a labeled acetate ester was synthesized. The labeled acetate ester co‐migrated with cis‐vaccenyl acetate in thin layer chromatography. Incubation of the abdomens of males from which the ejaculatory bulbs had been removed, or the abdomens of females, with radiolabeled acetate did not yield any acetate ester, but did yield other lipid products, including hydrocarbons. When the isolated labeled acetate ester was hydrolyzed, no radioactive vaccenol was formed. This strongly suggests that the acetyl group is incorporated via a transacetylation reaction, but that the vaccenyl moiety is not synthetized in the blub. The transacetylation enzyme activity was localized in the microsomal subfraction of the bulb homogenate, and its affinity for vaccenol was not very different from that reported for monounsaturated alcohol substrates in moths.     

The dehesa: an agrosilvopastoral system of the Mediterranean region with special reference to the Sierra Morena area of Spain

A multidisciplinary team undertook a six-year investigation on the agro-ecological and socio-economic aspects of the dehesa system in the Sierra Norte area, a part of the Sierra Morena of the Sevilla Province, Western Andalusia, Spain. This paper summarizes its findings on the various agro-ecological features of the system and evaluates the system's functional aspects vis-a-vis the trends and developments in land use in the region over the past few centuries. The synergistic effects of tree cover on understorey grassland vegetation is discussed in terms of soil fertility build-up and favourable micro-climatic and hydrological features, and the potential use of this information in future research and development programmes to improve the dehesa system in the Mediterranean region is outlined.     

Fruit load and branch ring-barking affect carbon allocation and photosynthesis of leaf and fruit of Coffea arabica in the field

Increasing fruit load (from no berries present to 25, 50 and 100% of the initial fruit load) significantly decreased branch growth on 5-year-old coffee (Coffea arabica L.) trees of the dwarf cultivar 'Costa Rica 95', during their third production cycle. Ring-barking the branches further reduced their growth. Berry dry mass at harvest was significantly reduced by increasing fruit load. Dry matter allocation to berries was four times that allocated to branch growth during the cycle. Branch dieback and berry drop were significantly higher at greater fruit loads. This illustrates the importance of berry sink strength and indicates that there is competition for carbohydrates between berries and shoots and also among berries. Leaf net photosynthesis (P(n)) increased with increasing fruit load. Furthermore, leaves of non-isolated branches bearing full fruit load achieved three times higher P(n) than leaves of isolated (ring-barked) branches without berries, indicating strong relief of leaf P(n) inhibition by carbohydrate demand from berries and other parts of the coffee tree when excess photoassimilates could be exported. Leaf P(n) was significantly higher in the morning than later during the day. This reduction in leaf P(n) is generally attributed to stomatal closure in response to high irradiance, temperature and vapor pressure deficit in the middle of the day; however, it could also be a feedback effect of reserves accumulating during the morning when climatic conditions for leaf P(n) were optimal, because increased leaf mass ratio was observed in leaves of ring-barked branches with low or no fruit loads. Rates of CO(2) emission by berries decreased and calculated photosynthetic rates of berries increased with increasing photosynthetic photon flux (PPF) especially at low PPFs (0 to 100 micromol m(-2) s(-1)). The photosynthetic contribution of berries at the bean-filling stage was estimated to be about 30% of their daily respiration costs and 12% of their total carbon requirements at PPF values commonly experienced in the field (200 to 500 micromol m(-2) s(-1)).     

Form and treatment effects on conical roundwood tested in bending

Summary This article presents the results and the comments of a sampling of 80 spruce conical roundwoods (tree trunks with the bark removed) of structural sizes tested in bending. Pressure impregnated samples were prepared with and without heart holes and paired with the untreated samples. Treatment effects have been studied in order to investigate their influences on the mechanical behavior. With a non-destructive evaluation by ultrasound, the mechanical results have been compared with an equivalent sawn timber series to analyze the effect of the form. The results show a negative influence of about 10% for the pressure-impregnated treatment on the mechanical properties only and up to 20% for the samples which were associated with pinholes for the heart treatment. On the other hand, the form effect increases the mechanical characteristics by 30% when compared to the square cross section. These results can be explained by a semi-ductile phase observed before failure. This conical roundwood form contributes to optimize the mechanical behavior linked to the anisotropic conditions where the grain is continuous and has a zero angle value with the longitudinal axis.