Comparison of three enzyme-linked immunosorbent assays for serologic diagnosis of contagious agalactia in sheep.

Serologic diagnosis of ovine contagious agalactia (Mycoplasma agalactiae) with the enzyme-linked immunosorbent assay (ELISA) developed by Agence Fran?aise de Sécurité Sanitaire des Aliments (AFSSA) may produce a few false-positive (FP) and false-negative (FN) results. When the prevalence of disease is low, these erroneous results may generate problems for eradication schemes. To prevent this, 2 commercial ELISAs were compared with the AFSSA ELISA. Flocks of known status were selected and classified into 4 categories: true positive (TP), FP, true negative (TN), and FN; 20 sheep per flock were submitted for blood sampling. A flock was considered positive when at least 1 out of 20 sera was positive or 2 sera were doubtful. In the flock, the diagnostic sensitivity of the 3 kits was very good (100%), and the diagnostic specificity showed an improvement from 46% (AFSSA test) to 88% and 92% (commercial tests). Considering individual animals, very few positive ewes were detected within TN or FP flocks; the proportion of positive ewes varied greatly from one kit to another (48% to 82%) within TP flocks. The kinetics of antibody response in sheep experimentally infected with various field strains of M. agalactiae were quite similar with all 3 ELISAs. The agreement between the 3 tests, assessed using the kappa value, varied from moderate to good (respective values of 0.56, 0.61, and 0.86). The 2 commercial ELISAs showed better performances, probably because of a superior analytical sensitivity, and are a good alternative for the serodiagnosis of contagious agalactia in sheep.     

The effect of chondrocyte cryopreservation on cartilage engineering

Chondrocytes were collected from the stifle joints of four pigs to study the effect of cryopreservation on the chondrogenic potential of chondrocytes. Half of the cells were cryopreserved for 3months. Polyglycolic acid scaffolds were cultured with fresh or cryopreserved chondrocytes for 4weeks. Cell morphology and the quality of engineered tissue were evaluated by scanning electron microscopy, histopathology and biochemical methods. More cells attached to scaffolds at 48h when fresh chondrocytes were seeded. At 4weeks, the numbers of cells, DNA and collagen II were greater in constructs engineered by fresh cells. However, the collagen II/DNA ratio did not differ between the two groups. More matrix was identified on a scanning electron microscope and by histopathology in the fresh group. Cartilage engineered with cryopreserved chondrocytes may contain less matrix and fewer cells. These findings most likely resulted from a lack of cell attachment on the matrix secondary to cryopreservation. Future studies are needed to further evaluate the mechanism by which cryopreservation may affect chondrocyte attachment.     

Doxorubicin area under the curve is an important predictor of neutropenia in dogs with naturally occurring cancers

Doxorubicin (DOX) area‐under‐the‐curve (AUC) was calculated for 40 dogs with spontaneously occurring cancers using a previously validated limited‐sampling approach. All dogs were administered a dose of 30 mg/m² by intravenous infusion and serum samples were collected at 5, 45 and 60 minutes post‐infusion. DOX and its major metabolite, doxorubicinol (doxol), were quantified in serum samples using high‐performance liquid chromatography tandem‐mass spectrometry. Wide interpatient variability was observed in the predicted DOX AUC with a coefficient of variation of 34%. A significant relationship was found between DOX AUC and absolute white blood cell count (P = 0.003), absolute neutrophil count (ANC; P = 0.002) and surviving fraction of neutrophils (P = 0.03) approximately 1 week after dosing (nadir). No changes in other hematologic parameters (red blood cells, platelets, lymphocytes, haemoglobin) were found to correlate with DOX AUC. The absolute dose (mg) and the dose per unit body weight (mg/kg) were not significantly correlated with nadir ANC. No relationships were found between maximum serum doxol concentration and myelosuppression. Baseline ANC was also significantly correlated to nadir ANC and a model was constructed using baseline ANC and DOX AUC that significantly described the nadir ANC. These findings demonstrate the important relationship between systemic DOX exposure and degree of neutropenia in dogs, and suggest a potential for individualized, pharmacokinetically‐guided DOX dosing in dogs.     

A decision support system to simulate and compare silvicultural scenarios for pure even-aged larch stands

In sustained forest management, it is particularly useful to test the adequacy of various silvicultural scenarios, but decision-making is also becoming increasingly complex because forest managers have to simultaneously meet several different objectives in response to society demands. In order to help forest managers make appropriate choices in silvicultural systems, we propose a SDSS (Silvicultural Decision Support System) that applies to pure and even-aged larch stands (Larix sp.) in lowland areas where site conditions are similar to those encountered in Southern Belgium. Its main purpose is (i) to predict the influence of silvicultural treatments on stand evolution and (ii) assist forest managers in comparing different scenarios with respect to predefined goals. This SDSS consists in three modules designed to elaborate silvicultural scenarios involving (i) stand growth prediction and thinning simulation; (ii) assessment of indicators selected to define the scenarios referring to wood production, financial, technico-economic and ecological components, tree stability and wood quality; and (iii) comparison of the scenarios on the basis of the set of previously assessed indicators (multi-criteria analysis). User-friendly “MGC_Larch” software helps managers to formulate, evaluate and compare different silvicultural scenarios for larch.     

Discovery of an amylose-free starch mutant in cassava (Manihot esculenta Crantz)

One of the objectives of the cassava-breeding project at CIAT is the identification of clones with special root quality characteristics. A large number of self-pollinations have been made in search of useful recessive traits. During 2006 harvests an S1 plant produced roots that stained brownish-red when treated with an iodine solution, suggesting that it had lower-than-normal levels of amylose in its starch. Colorimetric and DSC measurements indicated low levels (3.4%) and an absence of amylose in the starch, respectively. SDS-PAGE demonstrated the absence of GBSS enzyme in the starch from these roots. Pasting behavior was analyzed with a rapid visco-analyzer and resulted in larger values for peak viscosity, gel breakdown, and setback in the mutant compared with normal cassava starch. Solubility was considerably reduced, while the swelling index and volume fraction of the dispersed phase were higher in the mutant. No change in starch granule size or shape was observed. This is the first report of a natural mutation in cassava that drastically reduces amylose content in root starch.