Contribution to the reproductive ecology of Notoscopelus resplendens (Richardson, 1845) (Myctophidae) in the Central-Eastern Atlantic.

Notoscopelus resplendens is an abundant myctophid in the region of the Central-Eastern Atlantic. As with a majority of other myctophid species, this species performs vertical migration, playing a key role in the oceanic food web and in carbon sequestration. We examined the reproductive biology of N. resplendens based on 579 specimens caught between 1997 and 2002 off the Canary Islands. We found that the maximum standard length (SL) was lower than the size reported by other authors. The sex ratio was not different from 1:1. The average size at first maturity (L50) was higher in females (60.34 mm SL) than in males (56.61 mm SL). The gonadosomatic index (GSI) at 50% sexual maturity in females was higher than that in males. The reproductive activity was observed from January to April, while from May onwards, the majority of fish caught were in the process of maturation. The macroscopic scale of maturation was validated through the histological analysis of the ovarian development. The batch fecundity was related to the standard length, with an average of 1068.69 ± 369.84 eggs/spawn. These first data obtained for N. resplendens indicated that it is a batch spawner with asynchronous ovarian development.

Gastro-intestinal helminth parasites of the common North African gerbil Meriones shawi (Duvernoy) in Tunisia: Parasites diversity and habitat anthropization effect.

This study presents the first data on helminth fauna of Shaw's jird population from Tunisia. Ten helminth taxa were recovered from the digestive tract and the body cavity of 85 Shaw's jird Meriones shawi captured from two localities with different levels of anthropization in Tunisia. Among the helminth species recovered, only three have high prevalences and intensities, namely: Meggittina aegyptica, Gongylonema neoplasticum and Railliettina sp.. Helminth species richness from Mezzouna (less anthropized place) seems to be higher compare to Skhira (highly anthropized place). The nematode Trichuris gerbilli is more prevalent in Skhira (χ2=10.5, p=0.0012) and the cestodes Meggittina numida and M. aegyptica are more common in Mezzouna (χ2=9.6, p=0.0019; χ2=22.01, p<0.0001, respectively). This may be related to the high level of human activities which repulse wild animals, habitat fragmentation through the creation of agricultural parcels and their associated pollution by pesticides that kills insects which are intermediate hosts of helminths and discharges from the phosphate industry observed in Skhira. We add also a check-list of helminths species recorded from M. Shawi in all its distribution area.

Production of stable isotope labelled lipase Lip2 from Yarrowia lipolytica for NMR: investigation of several expression systems.

Extracellular lipase Lip2 from Yarrowia lipolytica is a promising biocatalyst with unusual structural features, as indicated by X-ray crystallography. These features comprise a mobile domain called the lid that controls access to the catalytic site. Conformational rearrangements of the lid have been suggested to regulate lipase enzymatic activities. We used nuclear magnetic resonance to investigate the dynamics of Lip2 by exploring four expression systems, Escherichia coli, cell-free, Pichia pastoris and Y. lipolytica to produce uniformly labelled enzyme. The expression of Lip2 was assessed by determining its specific activity and measuring (15)N-(1)H HSQC spectra. Y. lipolytica turned out to be the most efficient expression system. Here, we report the first use of Y. lipolytica as an expression host for the production of uniform stable isotopic labelled protein for further structural and dynamics studies using NMR.

Metabolic engineering for ricinoleic acid production in the oleaginous yeast Yarrowia lipolytica.

Although there are numerous oleochemical applications for ricinoleic acid (RA) and its derivatives, their production is limited and subject to various safety legislations. In an effort to produce RA from alternative sources, we constructed a genetically modified strain of the oleaginous yeast Yarrowia lipolytica. This strain is unable to perform ß-oxidation and is invalidated for the native triacylglycerol (TAG) acyltransferases (Dga1p, Dga2p, and Lro1p) and the ∆12 desaturase (Fad2p). We also expressed the Ricinus communis ∆12 hydroxylase (RcFAH12) under the control of the TEF constitutive promoter in this strain. However, RA constituted only 7% of the total lipids produced by this modified strain. By contrast, expression of the Claviceps purpurea hydroxylase CpFAH12 in this background resulted in a strain able to accumulate RA to 29% of total lipids, and expression of an additional copy of CpFAH12 drove RA accumulation up to 35% of total lipids. The co-expression of the C. purpurea or R. communis type II diacylglycerol acyltransferase (RcDGAT2 or CpDGAT2) had negative effects on RA accumulation in this yeast, with RA levels dropping to below 14% of total lipids. Overexpression of the native Y. lipolytica PDAT acyltransferase (Lro1p) restored both TAG accumulation and RA levels. Thus, we describe the consequences of rerouting lipid metabolism in this yeast so as to develop a cell factory for RA production. The engineered strain is capable of accumulating RA to 43% of its total lipids and over 60 mg/g of cell dry weight; this is the most efficient production of RA described to date.

Multigram-scale synthesis of an orthogonally protected 2-acetamido-4-amino-2,4,6-trideoxy-D-galactose (AAT) building block.

Reported is the gram-scale synthesis of tert-butyldiphenylsilyl 4-(N-benzyloxycarbonyl)-amino-2-azido-2,4,6-trideoxy-ß-D-galactopyranoside, which represents an orthogonally protected 2,4-diamino-D-fucose building block, a common constituent of various zwitterionic polysaccharides. The building block has been synthesized from D-glucosamine in 19% overall yield over 14 steps, requiring 5 chromatographic purifications. The key step in the synthesis is the introduction of the C-4 amino substituent, which has been accomplished by a one-pot three step procedure, involving regioselective C-3-O-trichloroacetimidate formation, C-4-O-triflation, and intramolecular substitution. The building block can be used as an acceptor and is readily transformed into a donor glycoside.

Improvement of Yarrowia lipolytica lipase enantioselectivity by using mutagenesis targeted to the substrate binding site.

Lip2p lipase from Yarrowia lipolytica was shown to be an efficient catalyst for the resolution of 2-bromo-arylacetic acid esters, an important class of chemical intermediates in the pharmaceutical industry. Enantioselectivity of this lipase was improved by site-directed mutagenesis targeted to the substrate binding site. To guide mutagenesis experiments, the three-dimensional model of this lipase was built by homology modelling techniques by using the structures of lipases from Rhizomucor miehei and Thermomyces lanuginosa as templates. On the basis of this structural model, five amino acid residues (T88, V94, D97, V232, V285) that form the hydrophobic substrate binding site of the lipase were selected for site-directed mutagenesis. Position 232 was identified as crucial for the discrimination between enantiomers. Variant V232A displayed an enantioselectivity enhanced by one order of magnitude, whereas variant V232L exhibited a selectivity inversion. To further explore the diversity, position 232 was systematically replaced by the 19 possible amino acids. Screening of this library led to the identification of the V232S variant, which has a tremendously increased E value compared to the parental enzyme for the resolution of 2-bromo-phenylacetic acid ethyl ester (58-fold) and 2-bromo-o-tolylacetic acid ethyl ester (16-fold). In addition to the gain in enantioselectivity, a remarkable increase in velocity was observed (eightfold increase) for both substrates.

Helminth species diversity of mammals: parasite species richness is a host species attribute.

Studies investigating parasite diversity have shown substantial geographical variation in parasite species richness. Most of these studies have, however, adopted a local scale approach, which may have masked more general patterns. Recent studies have shown that ectoparasite species richness in mammals seems highly repeatable among populations of the same mammal host species at a regional scale. In light of these new studies we have reinvestigated the case of parasitic helminths by using a large data set of parasites from mammal populations in 3 continents. We collected homogeneous data and demonstrated that helminth species richness is highly repeatable in mammals at a regional scale. Our results highlight the strong influence of host identity in parasite species richness and call for future research linking helminth species found in a given host to its ecology, immune defences and potential energetic trade-offs.

Eimeria (Coccidia: Eimeridea) of hares in France: description of new taxa.

The oocysts of coccidian of the genus Eimeria were sought in the caecal contents of 46 Lepus granatensis and 18 L. europaeus captured in France. Parasites were found in 34 of the hares. Parasite load was mainly very low. However, species diversity was considerable. 21 species or subspecies were identified, of which 13 species and two subspecies were not previously described. Three of the taxa, E. robertsoni, E. semisculpta and E. townsendi, previously identified on numerous occasions in western Europe and, corresponding to forms or variants created before 1960 that have been subsequently elevated to a specific level, appear to be invalid. Indeed, the parasite descriptions from the material used to effect this modification do not correspond to the original descriptions. A stable equilibrium, as generally observed in the case of many congeneric species co-infection of the same host, was not observed in the hares. This has been attributed to the solitary habits of the host and of the probable polyphyletic nature of the genus Eimeria. Paleontological data for the Leporidae indicate that rabbit parasites are derived from those of the hare.

[Should all prosthetic valve patients be treated by platelet inhibitors?]. / Faut-il traiter tous les porteurs de prothèses valvulaires par les inhibiteurs plaquettaires

The number, volume and adhesive and aggregation activities of platelets were studied in 31 patients with artificial valves. The differences observed in comparison with a control population are due to stimulation of thrombocytopoiesis secondary to the destruction or concumption of the platelets, it being impossible to be sure as to the dominant mechanism. Under these circumstances it would seem important in each patient to verify the therapeutic effectiveness of platelet inhibitors or of traditionnal anticoagulants by direct or indirect study of platelet life duration.