Rapid, low cost thin-layer chromatographic screening method for the detection of ochratoxin A in green coffee at a control level of 10 microg/kg.

A thin-layer chromatographic (TLC) screening method was developed for the detection of ochratoxin A (OTA) in green coffee at a control level of 10 microg/kg. The method is based on extraction of OTA with a mixture of phosphoric acid and dichloromethane, purification by liquid-liquid partition into sodium hydrogen carbonate, separation by normal-phase TLC, and detection by visual estimation of fluorescence intensity under a UV lamp at 366 nm. The method was validated by performing replicate analyses of uncontaminated green coffee material spiked at 3 different levels of OTA (5, 10, and 20 microg/kg), and also by comparing results obtained on a series of test trial green coffees naturally contaminated with OTA (range 0.2 to 136.7 microg/kg) with those measured by a quantitative immunoaffinity/HPLC method. The agreement between the two methods was excellent, and neither false positive nor false negative results were recorded. This screening method is rapid, simple, robust, and very cheap, which makes it particularly well adapted for implementation in coffee-producing countries.     

Development of a 16S rRNA microarray approach for the monitoring of rhizosphere Pseudomonas populations associated with the decline of take-all disease of wheat

So far, the analysis of microbial populations associated with wheat monocropping-induced decline of take-all disease (Gaeumannomyces graminis var. tritici) has focused mainly on culturable biocontrol pseudomonads. The objective of this study was to develop a taxonomic rrs (16S rRNA gene) microarray to assess the changes in Pseudomonas populations taking place during take-all decline. The microarray contains 12 probes for five Pseudomonas phylogenetic clusters chosen because they include well-known plant-beneficial pseudomonads. Four of the clusters are within the ‘Pseudomonas fluorescens’ species complex. PCR primers were selected to target these five clusters, and they were validated using 53 pseudomonads belonging or not to these clusters. Microarray analysis of the pseudomonads enabled discrimination between strains from several Pseudomonas clusters. Rhizosphere samples were collected from field plots grown with wheat for 1 (low level of take-all disease), 5 (high level of disease) or 10 years (low level of disease, suppressiveness reached). Microarray data could distinguish Pseudomonas populations from some of the wheat plants grown in the same plot. When comparing treatments, there was a difference between years 1 and 10. Cloning–sequencing of rrs enabled to define more precisely this difference by identifying two major Pseudomonas populations, one associated with year 1 and the other with year 10 (disease suppressiveness), which represent new clades within the ‘P. fluorescens’ complex. These populations may be useful as soil quality indicators. In conclusion, the combination of microarray and cloning–sequencing approaches highlighted changes in the prevalence of two major Pseudomonas populations, giving new insights on the dynamics of root-associated pseudomonads during take-all decline.     

<Emphasis Type="Italic">Allanblackia floribunda</Emphasis>: a new oil tree crop for Africa: amenability to grafting

Three Allanblackia species (A. floribunda, A. stuhlmannii and A. parviflora) with high nutritive, medicinal, cosmetic and economic values are currently being domesticated as new oil tree crops. Allanblackia seeds contain a hard white fat consisting mostly of stearic (52–58%) and oleic (39–49%) acids. This unusual fatty acid content has the right properties for many different food and cosmetic products making them commercially interesting. Vegetative propagation studies on A. floribunda, which grows naturally in the moist forest of Cameroon and Nigeria, were initiated aimed at evaluating its amenability to grafting. Scions were grafted onto 18 month old rootstocks of A. floribunda using side tongue, top cleft, side veneer, whip-and-tongue methods under nursery conditions in Cameroon. In parallel, side tongue and inverted ‘T’ budding methods were also tested in situ on young A. floribunda wildings growing under semi-deciduous and evergreen tree covers. In addition, the effects of protecting side tongue new grafts with non perforated translucent plastic, perforated translucent plastic and aluminium foil were assessed. Under nursery conditions, side tongue grafts were significantly more successful (80.0 ± 6.3%), than grafts of side veneer (52.5 ± 7.9%), top cleft (55.0 ± 7.9%) and whip and tongue (50.0 ± 7.9%). The success of side tongue graft was further increased (86.7 ± 6.2%) under the shade of evergreen trees when protected by non perforated translucent plastic. These results indicate the potential for in situ grafting and ‘top working’ to promote cultivation of more productive germplasm of Allanblackia within multifunctional agricultural systems.     

Ultrasound-guided serial transabdominal cardiac biopsies in cats

A percutaneous/transdiaphragmatic core needle biopsy technique was developed in cats to obtain serial biopsies from different locations of the left ventricle, through which morphological and molecular changes within the same individual can be studied to unravel the mechanisms of feline cardiomyopathies. Transmural left ventricular myocardial samples were obtained from 29 anesthetized, healthy, adult cats with ultrasound guidance. An 18G automatic biopsy needle was inserted between the last left rib and the sternum through the diaphragm into the thorax. Biopsies were obtained from the left ventricular wall. In five cats, three single biopsies were taken with 4-week intervals. Autopsy was performed on six cats, of which three cats had serial biopsies. In total, 87 biopsies were obtained without long-term effects on cardiac function or structure. The biopsies caused transient single ventricular premature complexes and mild pericardial effusion without tamponade. Necropsy revealed a minimal amount of fibrous connective tissue in the diaphragm and the heart without any significant microscopic lesions in the adjacent muscle tissue. The high quality biopsy material was suitable for morphological and molecular studies. This minimally invasive, ultrasound-guided cardiac biopsy technique thus allows for the safe collection of serial biopsies to study feline cardiomyopathies in an experimental setting.     

Stabilization of aroma compounds through sorption-release by packaging polymers

Plastic packaging materials are often associated to aroma losses and to a decrease of the organoleptic quality of foods. This work defines situations where, on the contrary, plastics play a regulating role on the concentration of reactive aroma compounds in foods. These systems can be described by a two step mechanism; first, aroma is sorbed in the polymer, while the fraction in solution degrades quickly; in a second step, as the concentration is close to zero in the solution, the polymer liberates progressively the sorbed aroma back to the food. A simple numerical model is proposed, describing competitive processes of aroma degradation in solution and sorption by a polymer in contact with a homogeneous aqueous food. The classical limonene/low density polyethylene (LDPE) system is studied experimentally for the validation of the model: in an acidic medium, limonene both degrades quickly and is sorbed quickly, with a large solubility in LDPE. To define which aroma packaging systems could also display this interesting behavior, all types of possible interactions, using thermodynamic and kinetic parameters describing most practical situations, are simulated. For that purpose, 35 values of reference diffusion coefficients and 35 partition coefficients of usual aroma compounds between polymers and water have been measured and combined with the few available data from literature. The situations where polymers regulate the aroma concentration in food correspond to large partition coefficients (above 10), large diffusion coefficients (>10(-9) cm2 x s(-1)), and large degradation constants.     

Keratoprosthesis with retrocorneal fixation: preliminary results in dogs with corneal blindness

Objective To evaluate the use and complications of a penetrating keratoprosthesis implantation in the management of corneal opacification in dogs. Methods A retrospective clinical study describes the indications for the surgical technique utilized and the outcomes of this procedure in 20 eyes of 19 dogs with blindness of corneal origin. A successful surgical outcome was defined as a clear keratoprosthesis optic and improvement or restoration of functional vision over a follow‐up period ranging from at least 8 months to a maximum of 7 years. Results Eyes with total corneal opacification resulting from chronic superficial keratitis (n = 11), keratoconjunctivitis sicca (n = 5), endothelial dystrophy (n = 3) and chemical burn (n = 1) were treated by unilateral (n = 18) or bilateral (n = 1) full‐thickness implantation of a keratoprosthesis. Keratoprostheses were retained in 15 eyes (75%) which regained vision to the date of reporting. Among these eyes, six had uncomplicated postoperative course, five developed retroprosthetic membranes and four developed granulation tissue over the optic of the keratoprostheses. These complications were successfully removed surgically in the nine eyes. The five remaining eyes (25%) developed serious early postoperative complications, for which enucleation had to be performed. Conclusion In keratopathies in which the corneal opacification could not be treated by standard medical or surgical procedures, this keratoprosthesis appears to be promising to restore vision in chronic superficial keratitis and deep corneal dystrophy. It appears to have a poor prognosis in keratoconjunctivitis sicca in brachycephalic dogs. The post operative complications retro‐prosthetic membranes and granulomatous overgrowth could be treated well.