3D Reconstruction of the Mouse's Mesonephros

The present work reports on the three-dimensional reconstruction of the segmented mesonephros during the embryonic development of the mouse. With a light microscope and an automatic reconstruction of surfaces, aspects of the mesonephros are described. These surfaces are obtained by using digitized contour lines. A new interpolation method called DSI (Discrete Smooth Interpolation) enables correction of the distortion induced by microtomy in paraffin sections. After a tri-angulation step, this method uses a smoothing algorithm, which implies a spatial redistribution of the vertices of the triangles to correct the rotational and translational misalignment. The use of this 3D program improves the understanding of the development patterns and helps us to appreciate changes in the rebuilt mesonephros. By 10.5 embryonic days, tubules emerge from the Wolffian body and begin their formation, then between 11.5 and 13.5 embryonic days, tortuous mesonephric tubules bound to the Wolffian duct form small curls, which grow and finally unwind. At the same time, mesonephric tubules unbound to the Wolffian duct appear, and on 13.5 embryonic days, the Müllerian duct is visible. After 14.5 embryonic days, the segmented mesonephros keeps its general aspect but decreases in size. At this time, each gonad is provided with both Wolffian and Müllerian ducts. Later, the Wolffian duct differentiates into the definitive male duct system, whilst the Müllerian duct regresses. Conversely, the paramesonephric duct differentiates into the definitive female duct system, whilst the mesonephric duct in turn degenerates. By this time degeneration has begun in the cranial portion of the mesonephros and this process progresses caudally. The spatial organization of the mesonephric tubules and the precise organization of all connections between these elements and the ducts may be well defined. Such approach can allow for a high definition of the normal pattern of mesonephros differentiation.     

Enhanced viability of a nervous necrosis virus-infected stable cell line over-expressing a fusion product of the zfBcl-xL and green fluorescent protein genes

Nervous necrosis virus (NNV) infection induces host cell apoptosis by an ill-understood process. We utilized a fusion between enhanced green fluorescent protein (EGFP) and the zfBcl-x(L) gene in GL-av cells to select for zfBcl-x(L) stable cell lines and to assess the effectiveness of the anti-apoptotic protein Bcl-x(L) in circumventing NNV-induced cell death. Stable EGFP and EGFP-Bcl-x(L)-expressing clones were obtained at high purity within 2.5-3 months. In the latter, the EGFP-Bcl-x(L) fusion protein (approximately 58.2 kDa, as ascertained by Western blot) was predominantly targeted to mitochondria. We assayed for apoptosis in red-spotted grouper NNV Tainan no. 1 (RGNNV TN1)-infected cells with terminal deoxynucleotidyl transferase (TdT)-mediated end labelling (TUNEL) of DNA at different virus doses. NNV infection of NNV Bcl-x(L) GL-av cell line revealed a protective effect, with a decrease in TUNEL-positive cells of 7%, 8% and 31.8% at 24, 48 and 72 h, respectively. In addition, RGNNV infection of the Bcl-x(L) GL-av cell line revealed a protective effect, with an enhanced viability of 3%, 40% and 73% at 24, 48, and 72 h, respectively. We conclude that NNV-induced apoptotic cell death can be lessened in transgenic grouper fish cells.     

Effects of the initial level of milk production and of the dietary protein intake on the course of natural nematode infection in dairy goats

The level of dietary protein is known to alter the establishment and the pathophysiology of gastrointestinal nematodes in sheep. On the other hand, high-producing dairy goats are less resistant and/or resilient than low-producing ones to experimental nematode infection. During a 2-year study, we have investigated the course of a natural nematode infection (mainly T. colubriformis) in a high-producing dairy goat flock. In year 1, 50 grazing goats divided in high (HP) or low producer (LP) were compared from April to October for parasitological and milk parameters. In year 2, the 25 HP goats were only considered and were allocated to two levels of dietary protein, high level (HPr) with a protein coverage of 125% or normal level (NPr) with a protein coverage of 106%. They were monitored as above. In year 1, HP goats showed a greater nematode egg output (1856 vs. 1000epg) associated with higher values in T. colubriformis IgG in autumn than LP ones whereas the decrease in serum phosphate concentration was similar in both groups. In year 2, HPr goats exhibited lower egg output in autumn (2219 vs. 2817) vs. NPr ones. Moreover, milk yield and fat content were significantly higher in HPr vs. NPr in the 2nd part of the study. HP goats are less resistant to nematode infection in natural conditions. Resistance and resilience of HP goats may partially be improved by a protein supplementation in the diet.     

Effects of dietary scFOS on immunoglobulins in colostrums and milk of bitches

Fructo-oligosaccharides (scFOS) are prebiotic ingredients that improve protection against pathogens probably through promoting the growth of gastrointestinal bacteria-like Bifidobacteria and Lactobacilli: this stimulation may lead to a better development of immune repertoire and/or stimulation of the local immune response. According to the existence of the immune entero-mammary link, we were wondering if the dietary supplementation with scFOS could enhance the mucosal immunoglobulin level in mammary secretions. Results in this study show that bitches supplemented with scFOS exhibit higher colostrum and milk IgM content without concomitant effect on IgG1, IgG2 and IgA. In addition, intranasally immunized puppies exhibited a trend to higher Bordetella bronchiseptica-specific IgM immune response. The dietary supplementation with scFOS increased the IgM level in colostrums and milk of bitches by mechanisms which remain to be elucidated.     

Comparison of eggshell hygiene in two housing systems: standard and furnished cages

1. The hygienic properties of eggs produced in two designs of furnished cage were investigated and compared to two standard cage designs. 2. At 28, 37, 47 and 58 weeks of age, the proportion of dirty eggs was higher in one of the furnished cage designs while, in the other, it was similar to standard cages. 3. At 27, 33 and 60 weeks, the bacterial load on the eggshell (total aerobic bacteria and enterococci) was higher in furnished cage designs. A seasonal effect was observed with lower counts at 60 weeks (winter) than at 27 weeks (summer). 4. More dirty eggs and a higher bacterial load were observed in eggs laid outside the nests, which suggests egg hygiene in furnished cages could be similar to standard cages if the equipment in furnished cages was improved to enhance nest laying.     

Improvement of nitrogen fixation in Acacia mangium through inoculation with rhizobium

Acacia mangium, a N₂‐fixing tree legume, has become a major plantation tree species in the tropical humid and sub-humid zones. In addition to being a major pulp-wood producer, the tree has a good potential to restore soil fertility as a fallow species in agroforestry systems, and as a fuel species. In this paper, we report an overview of the results from several rhizobium inoculation field trials in different edaphic and ecological conditions, conducted by the CIRAD-Forêt (The Forest Program of Centre de Coopération Internationale en Recherche Agronomique pour le Développement) in partnership with national research organizations of various countries of the humid and sub-humid lowlands of West Africa and Cook Islands. Rhizobium inoculation had a positive effect on tree growth up to 39 months after tree planting. Immunological identification of the Bradyrhizobium strains present in the nodules confirmed the persistence of the more efficient introduced strains up to 42 months after transfer of the inoculated trees to the field. In Côte d'Ivoire, nitrogen derived from atmospheric N₂ fixed symbiotically by A. mangium was 50% in the whole trial and up to 90% in plots with less fertile soils when the trees were inoculated with an efficient strain.