Use of a serum-free medium to produce in vitro Neospora caninum and Toxoplasma gondii tachyzoites on Vero cells

Neospora caninum and Toxoplasma gondii are cyst-forming coccidian parasites of human and veterinary clinical relevance. In vitro cultivation of the protozoans using Vero cells is usually performed in order to produce antigenic materials. Quantitative and qualitative comparisons of Vero cells grown in RPMI medium supplemented either with foetal calf serum (FCS), horse serum (HS) or a specific serum-free additive (DefCell) were performed. A serum-free cell culture system used to propagate N. caninum (NC-1 isolate) and T. gondii tachyzoites (Rh stain) were compared with the other two cell culture systems. FCS supplemented media was found to be more effective than the others in promoting Vero cells and N. caninum tachyzoites. However, it was found unable to support adequate T. gondii tachyzoite proliferation. Vero cells, T. gondii and N. caninum tachyzoite production gave similar growth patterns with either HS or DefCell supplemented media. Defcell was considered as a good alternative to supplement culture medium.     

Implication of Systemic Induced Resistance in the Suppression of Fusarium Wilt of Tomato by Pseudomonas fluorescens WCS417r and by Nonpathogenic Fusarium oxysporum Fo47

Fluorescent pseudomonads and nonpathogenic Fusarium oxysporum have been shown to suppress fusarium wilts. This suppression has been related to both microbial antagonism and induced resistance.The aim of the present study was to assess the relative importance of systemic induced resistance in the suppression of fusarium wilt of tomato in commercial-like conditions by a reference strain of each type of microorganism (P. fluorescens WCS417r and nonpathogenic F. oxysporum Fo47). The spatial separation of the pathogen and the biocontrol strains excluded any possible microbial antagonism and implicated the involvement of the systemic induced resistance; whereas the absence of any separation between these microorganisms allowed the expression of both mechanisms. Since systemic induced resistance has often been associated with the synthesis of PR-proteins, their accumulation in tomato plants inoculated with WCS417r or with Fo47 was determined.The analysis of the results indicates that the suppression of fusarium wilt by P. fluorescens WCS417r was ascribed to systemic induced resistance without any detection of the PR-proteins tested (PR-1 and chitinases). In contrast, the suppression achieved by nonpathogenic F. oxysporum Fo47 appeared to be mainly ascribed to microbial antagonism but also to a lesser extent to systemic induced resistance. This induced resistance could be related to the accumulation of PR-1 and chitinases.The possible relationship between the ability of Fo47 to suppress fusarium wilt more efficiently than WCS417r and its ability to show both mechanisms is discussed.     

Inactivation kinetics of purified tomato polygalacturonase by thermal and high-pressure processing

Tomato polygalacturonase (PG) was extracted from ripe tomatoes and purified by cation exchange and gel filtration chromatography. Cation exchange chromatography yielded two peaks with PG activity: the first peak was identified as PG2 (the heat labile form) and the second one as PG1 (the heat stable form). Both PG2 and PG1 presented a molar mass of 42 kDa when analyzed by SDS-PAGE and an isoelectric point >9.3. Thermal inactivation of purified tomato PG2, at pH 4.4, in the temperature range from 53 to 63 degrees C, followed first-order kinetics. Combined pressure-temperature inactivation of tomato PG2 was studied at 5-55 degrees C/100-600MPa. Under all pressure-temperature conditions, PG2 inactivation followed first-order kinetics. Purified tomato PG1, although more thermostable than PG2, showed a pressure stability very similar to that of PG2. These results indicate that high-pressure processing is an efficient alternative to inactivate tomato PG without the need for applying high temperatures.     

Valorization of Brazilian vetiver (Vetiveria zizanioides (L.) Nash ex Small) oil

The valorization of extracts from Brazilian vetiver (Vetiveria zizanioides (L.) Nash ex Small) roots was studied. This study took into account the extraction method, the chemical composition of the extracts, their sensorial characteristics, and the possibility of chemical transformations of the product. The performed extraction methods were hydrodistillation and extraction with supercritical carbon dioxide. Some pretreatment methods were tested on the vetiver roots and evaluated in terms of extraction yield, process time, chemical composition, and sensorial properties. Supercritical carbon dioxide extraction resulted in high yield (3.2%) in significantly less time than the other methods. The chemical compositions of the extracts obtained by the different methods were also compared to those of commercial vetiver oils from other sources, showing that Brazilian samples had a greater acid amount. An extraction in basic medium from Brazilian vetiver oil was done to remove its main acid (zizanoic acid), which was chemically transformed into an alcohol (khusimol) of desirable sensorial properties. Sensory evaluation indicated that the Brazilian volatile oil without acid could be used in perfumery and the extract obtained with supercritical carbon dioxide could have application in food.     

Effects of trimethoprim-sulfadiazine on tear production and the fluctuations of Schirmer tear test values in horses

The objectives of this study were to observe the effects of trimethoprim-sulfadiazine on equine tear production and to determine normal fluctuations in Schirmer tear test (STT) values in horses. A randomized, placebo-controlled, blinded clinical trial measuring STT values in 15 horses over an 8-week period was performed. The treatment group (eight horses) received 30 mg/kg trimethoprim-sulfadiazine orally once a day and the control group (seven horses) received placebo (flour) at the same time. All horses were housed outdoors throughout the study. Schirmer tear test values were measured at 0, 2, 4, 6 and 8 weeks, and 4 weeks after discontinuation of treatment. There were no significant differences in tear production between the treated and control groups. Fluctuations in STT were observed and may result from individual and environmental variations. Trimethoprim-sulfadiazine did not decrease tear production in the horses in this study. Horses normally experience periodic fluctuations in STT values.     

Comparison of growth,digestive system maturation and skeletal development in sea bass larvae reared in an intensive or a mesocosm system

The quality of development in intensive or mesocosm hatchery‐reared Dicentrarchus labrax larvae was investigated using physiological indicators assessing ontogeny. Larvae were reared in intensive (120 L tanks) and in mesocosm systems (20 m³ enclosures) with the same feeding sequence, excluding the wild zooplankton naturally available in mesocosms. Faster growth was recorded since early development [16 day after hatching (DAH)] in the mesocosm. Maturation of the digestive system also occurred earlier as indicated by the higher amylase secretion ratios, the intestinal maturation index (alkaline phosphatase/leucine–alanine peptidase and aminopeptidase‐N/leucine–alanine peptidase ratios) and the more developed intestinal epithelium at 23 DAH. Nevertheless, the delay in digestive maturation in the intensive system seemed retrieved within few days. In both the groups, the number of vertebrae ranged between 24 and 26, with the dominant class being 25 vertebrae. However, the distributions differed between treatments for meristic characteristics, ossification stages and incidence of malformation types. Loss of a vertebra was more frequent in the intensive system, while the appearance of an additional vertebra was more frequent in the mesocosm. Ossification at 37 DAH was also more advanced in the mesocosm in addition to a lesser rate and severity of skeletal malformations. It is suggested that the early nutritional contribution of mesocosm wild zooplankton, yet at densities of 0.2–0.7 prey mL^?1, had key effects on larvae development since the early stages.     

Effect of soil nitrogen supply on carbon assimilation by tree stems

• ? Nitrogen (N) is one of the most important resources for plants, generally enhancing leaf photosynthesis because a large part of it is allocated to Rubisco and thylakoïds. This is well known in leaves where photosynthesis (i.e. gas exchange, Rubisco activity, chlorophyll content) is positively correlated to leaf N content.
• ? In order to test this hypothesis in stems, N concentration, CO₂ exchange and also Rubisco and PEP carboxylase activities were measured in summer on current-year stems of young European beeches (Fagus sylvatica L.) growing on soils of different N content.
• ? The CO₂ refixation rate of stems increased from 58.5% to 74.3% when stem N concentration increased from 5.7 to 10.1 mg g^?1 DW. A hyperbolic relationship was obtained between stem gross photosynthesis and N concentration, with an x-intercept of 0.3 mmol N g^?1 DW. Stem PEP carboxylase activity was higher in stems than in leaves and increased with stem N concentration whereas Rubisco activity did not change between treatments in both tissues.
• ? In spite of a low nitrogen investment in stem photosynthesis (low PNUE), these results suggest that (1) stems invest more N in CO₂ refixation when more N is widely available, (2) stem photosynthesis is able to operate at low N concentration and (3) stem PEP carboxylase is involved in stem carbon refixation, but also simultaneously supplies carbon skeletons for N assimilation.

     

Biotransformation of s-Triazine Herbicides and Related Degradation Products in Liquid Cultures by the White Rot Fungus Phanerochaete chrysosporium

The ability of the white rot basidiomycete Phanerochaete chrysosporium to transform s-triazine herbicides has been investigated in laboratory experiments. The chlorinated metabolites formed during atrazine N-dealkylations were not further transformed by the fungus, whereas hydroxy-atrazine was converted to an unknown product. P. chrysosporium was also able to carry out the N-dealkylation of the herbicides simazine, propazine and terbuthylazine. Herbicide metabolism was not supported by purified peroxidases. The highest rates of herbicide N-dealkylation were obtained in liquid cultures maintained under moderate temperature allowing a long mycelium growing phase. Atrazine transformation was found to be supported by the mycelium, which contained significant amounts of microsomal cytochrome P450. Herbicide N-dealkylation was decreased in the presence of 1-aminobenzotriazole, in agreement with the involvement of P450 monooxygenases in atrazine metabolism. © 1997 SCI.