Influence des gousses d''arachide (Arachis Hypogea) et de l''alimentation imaginale sur l''ovogénèse, l''accouplement et la ponte chez la bruche Caryedon Serratus

Résumé L'activité reproductrice d'une souche de Caryedon serratus provenant de stocks d'arachide a été étudiée au Congo dans des conditions de température et d'hygrométrie simulant celles d'un grenier paysan ainsi qu'à la température constante de 30°C. L'ovogénèse est stimulée, quoique faiblement, par la présence de gousses de la plante hôte. La présence du mâle est sans effet sur l'ovogénèse. La présence de gousses d'arachide n'a pas d'influence sur la copulation; la femelle est généralement réceptive dès qu'un premier ovocyte est parvenu à maturité. La ponte est fortement stimulée par la présence d'une gousse d'arachide. La rétention ovocytaire est très faible chez les femelles inséminées, même en l'absence de gousse. On observe chez les femelles vierges une rétention des ovocytes dans les oviductes latéraux, qui s'accompagne apparemment d'une résorption partielle; environ un tiers des ufs est néanmoins émis. A 30°C, des femelles régulièrement nourries de pollen présentent une fécondité moyenne d'environ 650 ufs; le maximum observé est de 1457 ufs. La longévité moyenne de ces femelles est d'environ trois mois, mais certaines vivent près de six mois, en l'absence de toute diapause.

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Summary Reproductive activity in a strain of Caryedon serratus originating from groundnut stores was studied in Congo under humidity, temperature and daylength conditions similar to those prevailing in farmers' stores as well as at constant 30°C. Oogenesis was slightly stimulated in the presence of unshelled groundnuts. The presence of males did not affect oogenesis. Groundnuts had no effect on mating. Females usually became sexually receptive after a first oocyte had matured. Oviposition by virgin females was strongly stimulated by the presence of nuts. Oocyte retention was very low in inseminated females, even in the absence of groundnuts. Virgin females exhibited oocyte retention in lateral oviducts, apparently together with resorption. Egg chorions were occasionnally observed in the bursa copulatrix of old virgin females, but no proof is given of the existence of resorption mechanisms in that gland. About one third of the eggload was eventually laid. At 30°C, the fecundity of inseminated and regularly pollen-fed females was about 650 eggs. Maximum lifetime egg production was 1457. These females had a mean length of life of about three months, and some individuals survived almost six months, with no sign of diapause.

     

Nutrition et élevage du prédateur polyphageMacrolophus caliginosus [Heteroptera,Miridae] sur milieux artificiels

La possibilité de nourrir la punaiseMacrolophus caliginosus Wagner à l'aide de milieux artificiels a été testée. En l'absence de végétal, les témoins nourris avec des ?ufs d'Ephestia kuehniella Zeller ont une survie médiocre et fournisent 33% d'adultes. Avec le meilleur milieu artificiel la survie est comparable à celle des témoins et 21 % d'adultes sont obtenus. Avec ce même milieu, mais en présence de feuille de géranium, la production d'adultes atteint 62%. Les compositions en acides aminés des punaises élevées avec le milieu montrent des écarts inférieurs à 20% par rapport aux témoins. Le développement complet deM. caliginosus nourri à l'aide de milieu artificiel a été obtenu. Le végétal joue un rôle important dans la biologie de ce prédateur.     

Pseudoperichaeta insidiosa un nouveau tachinaire[DIP.] élevé dansGalleria mellonella [Lep.]

Résumé Pseudoperichaeta insidiosa R.D., tachinaire parasite non spécifique d'Ostrinia nubilalis Hübner est élevée pour la première fois dans les larves deGalleria mellonella L. La méthode d'élevage est exposée en détail et certaines caractéristiques biologiques du développement post-embryonnaire sur cet h?te de substitution sont définies. Quelques éléments de la biologie des adultes au laboratoire sont décrits.

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Summary For the first time,Pseudoperichaeta insidiosa R.D., a non specific tachinid parasite ofOstrinia nubilalis (Hübner), was reared onGalleria mellonella L. With this substitution host a yield of about 0.35 pupa per deposited planidium is obtained. The mean pupal weight is 28.8 mg with 1 planidium per host and 25.0 with 2. Some post embryonic development and adult biological features are given. The whole life cycle lasts about 45 days. Till now 8 generations have been reared.

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Avec la collaboration technique deColette Ogier.     

Genetic analysis of Pycr1 and Pycr2 in mice

The final step in proline biosynthesis is catalyzed by three pyrroline-5-carboxylate reductases, PYCR1, PYCR2, and PYCR3, which convert pyrroline-5-carboxylate (P5C) to proline. Mutations in human PYCR1 and ALDH18A1 (P5C Synthetase) cause Cutis Laxa (CL), whereas mutations in PYCR2 cause hypomyelinating leukodystrophy 10 (HLD10). Here, we investigated the genetics of Pycr1 and Pycr2 in mice. A null allele of Pycr1 did not show integument or CL-related phenotypes. We also studied a novel chemically-induced mutation in Pycr2. Mice with recessive loss-of-function mutations in Pycr2 showed phenotypes consistent with neurological and neuromuscular disorders, including weight loss, kyphosis, and hind-limb clasping. The peripheral nervous system was largely unaffected, with only mild axonal atrophy in peripheral nerves. A severe loss of subcutaneous fat in Pycr2 mutant mice is reminiscent of a CL-like phenotype, but primary features such as elastin abnormalities were not observed. Aged Pycr2 mutant mice had reduced white blood cell counts and altered lipid metabolism, suggesting a generalized metabolic disorder. PYCR1 and -2 have similar enzymatic and cellular activities, and consistent with previous studies, both were localized in the mitochondria in fibroblasts. Both PYCR1 and -2 were able to complement the loss of Pro3, the yeast enzyme that converts P5C to proline, confirming their activity as P5C reductases. In mice, Pycr1; Pycr2 double mutants were sub-viable and unhealthy compared to either single mutant, indicating the genes are largely functionally redundant. Proline levels were not reduced, and precursors were not increased in serum from Pycr2 mutant mice or in lysates from skin fibroblast cultures, but placing Pycr2 mutant mice on a proline-free diet worsened the phenotype. Thus, Pycr1 and -2 have redundant functions in proline biosynthesis, and their loss makes proline a semi-essential amino acid. These findings have implications for understanding the genetics of CL and HLD10, and for modeling these disorders in mice.     

Macrophage-specific responses to human- and animal-adapted tubercle bacilli reveal pathogen and host factors driving multinucleated cell formation

The Mycobacterium tuberculosis complex (MTBC) is a group of related pathogens that cause tuberculosis (TB) in mammals. MTBC species are distinguished by their ability to sustain in distinct host populations. While Mycobacterium bovis (Mbv) sustains transmission cycles in cattle and wild animals and causes zoonotic TB, M. tuberculosis (Mtb) affects human populations and seldom causes disease in cattle. The host and pathogen determinants underlying host tropism between MTBC species are still unknown. Macrophages are the main host cell that encounters mycobacteria upon initial infection, and we hypothesised that early interactions between the macrophage and mycobacteria influence species-specific disease outcome. To identify factors that contribute to host tropism, we analysed blood-derived primary human and bovine macrophages (hMϕ or bMϕ, respectively) infected with Mbv and Mtb. We show that Mbv and Mtb reside in different cellular compartments and differentially replicate in hMϕ whereas both Mbv and Mtb efficiently replicate in bMϕ. Specifically, we show that out of the four infection combinations, only the infection of bMϕ with Mbv promoted the formation of multinucleated giant cells (MNGCs), a hallmark of tuberculous granulomas. Mechanistically, we demonstrate that both MPB70 from Mbv and extracellular vesicles released by Mbv-infected bMϕ promote macrophage multinucleation. Importantly, we extended our in vitro studies to show that granulomas from Mbv-infected but not Mtb-infected cattle contained higher numbers of MNGCs. Our findings implicate MNGC formation in the contrasting pathology between Mtb and Mbv for the bovine host and identify MPB70 from Mbv and extracellular vesicles from bMϕ as mediators of this process.     

Evidence of Coat Color Variation Sheds New Light on Ancient Canids

We have used a paleogenetics approach to investigate the genetic landscape of coat color variation in ancient Eurasian dog and wolf populations. We amplified DNA fragments of two genes controlling coat color, Mc1r (Melanocortin 1 Receptor) and CBD103 (canine-β-defensin), in respectively 15 and 19 ancient canids (dogs and wolf morphotypes) from 14 different archeological sites, throughout Asia and Europe spanning from ca. 12 000 B.P. (end of Upper Palaeolithic) to ca. 4000 B.P. (Bronze Age). We provide evidence of a new variant (R301C) of the Melanocortin 1 receptor (Mc1r) and highlight the presence of the beta-defensin melanistic mutation (CDB103-K locus) on ancient DNA from dog-and wolf-morphotype specimens. We show that the dominant K^B allele (CBD103), which causes melanism, and R301C (Mc1r), the variant that may cause light hair color, are present as early as the beginning of the Holocene, over 10 000 years ago. These results underline the genetic diversity of prehistoric dogs. This diversity may have partly stemmed not only from the wolf gene pool captured by domestication but also from mutations very likely linked to the relaxation of natural selection pressure occurring in-line with this process.     

Physical mapping and cloning of RAD56

Here we report the physical mapping of the rad56-1 mutation to the NAT3 gene, which encodes the catalytic subunit of the NatB N-terminal acetyltransferase in Saccharomyces cerevisiae. Mutation of RAD56 causes sensitivity to X-rays, methyl methanesulfonate, zeocin, camptothecin and hydroxyurea, but not to UV light, suggesting that N-terminal acetylation of specific DNA repair proteins is important for efficient DNA repair.     

Edesign: Primer and Enhanced Internal Probe Design Tool for Quantitative PCR Experiments and Genotyping Assays

Analytical PCR experiments preferably use internal probes for monitoring the amplification reaction and specific detection of the amplicon. Such internal probes have to be designed in close context with the amplification primers, and may require additional considerations for the detection of genetic variations. Here we describe Edesign, a new online and stand-alone tool for designing sets of PCR primers together with an internal probe for conducting quantitative real-time PCR (qPCR) and genotypic experiments. Edesign can be used for selecting standard DNA oligonucleotides like for instance TaqMan probes, but has been further extended with new functions and enhanced design features for Eprobes. Eprobes, with their single thiazole orange-labelled nucleotide, allow for highly sensitive genotypic assays because of their higher DNA binding affinity as compared to standard DNA oligonucleotides. Using new thermodynamic parameters, Edesign considers unique features of Eprobes during primer and probe design for establishing qPCR experiments and genotyping by melting curve analysis. Additional functions in Edesign allow probe design for effective discrimination between wild-type sequences and genetic variations either using standard DNA oligonucleotides or Eprobes. Edesign can be freely accessed online at http://www.dnaform.com/edesign2/, and the source code is available for download.     

Production of Mucosally Transmissible SHIV Challenge Stocks from HIV-1 Circulating Recombinant Form 01_AE env Sequences

Simian-human immunodeficiency virus (SHIV) challenge stocks are critical for preclinical testing of vaccines, antibodies, and other interventions aimed to prevent HIV-1. A major unmet need for the field has been the lack of a SHIV challenge stock expressing circulating recombinant form 01_AE (CRF01_AE) env sequences. We therefore sought to develop mucosally transmissible SHIV challenge stocks containing HIV-1 CRF01_AE env derived from acutely HIV-1 infected individuals from Thailand. SHIV-AE6, SHIV-AE6RM, and SHIV-AE16 contained env sequences that were >99% identical to the original HIV-1 isolate and did not require in vivo passaging. These viruses exhibited CCR5 tropism and displayed a tier 2 neutralization phenotype. These challenge stocks efficiently infected rhesus monkeys by the intrarectal route, replicated to high levels during acute infection, and established chronic viremia in a subset of animals. SHIV-AE16 was titrated for use in single, high dose as well as repetitive, low dose intrarectal challenge studies. These SHIV challenge stocks should facilitate the preclinical evaluation of vaccines, monoclonal antibodies, and other interventions targeted at preventing HIV-1 CRF01_AE infection.