Phenotypic effects of antibiotic‐induced axenity and egg disinfection in early larval European seabass (Dicentrarchus labrax L.)

In the present study, the hypothesis that the difference in axenic conditions in the incubation and rearing environment of European seabass larvae induces size and shape effects on the specimens is tested. This difference is studied between xenic and axenic seabass larvae of DAH (day after hatching) 0, 5, 11 and 15. The axenic rearing protocol involves an egg disinfection with glutaraldehyde after the primary one with iodine in the hatchery, and the hypothesis that this secondary disinfection induces size and shape effects is also tested. In order to accomplish this, three egg and larvae treatments are included: “DA” (disinfected axenic), “DX” (disinfected xenic) and “NX” (nondisinfected xenic). Regarding the effect of antibiotic‐induced axenity, DA larvae exhibited larger bodies than both DX and NX on DAH 5 and 11. They also had a smaller yolk sac than DX at hatching, but consumed it slower. Towards the end of the experiment, DA larvae were thicker, but slightly more curved than DX and NX, which may be an abnormal shape, or a slightly more advanced ontogenetic stage. As far as egg disinfection, it had significant but very moderate shape effects on DAH 5 and 11, and disinfected larvae consumed their yolk sac faster. To the best of our knowledge, this study is one of the first to illustrate the subtle but significant size and shape effects caused by antibiotic‐induced axenity and secondary egg disinfection in the early larval stages, which suggest the existence of bacterial mechanisms that play a phenotypic role.     

1H‐NMR metabolomic profiling of the crayfish Astacus leptodactylus subjected to polyphenol‐enriched diets

¹H‐NMR analysis of the hepatopancreas, muscle and haemolymph of Astacus leptodactylus after feeding with polyphenol‐enriched diet is reported. ¹H‐NMR spectra of lipophilic extracts showed the presence of cholesterol, fatty acid residues, phospholipids and triglycerides. ¹H‐NMR spectra of aqueous extracts identified 35 metabolites in the hepatopancreas, 31 in the muscle and 22 in the haemolymph. A total of 20 metabolites (amino acids and their derivatives) were present in the hepatopancreas, the muscle and the haemolymph. A total of 10 metabolites were present in both the hepatopancreas and the muscle (five amino acids, 2‐hydroxybutyrate, choline, myo‐inositol, glycogen and uracil). 2‐Hydroxyisobutyrate and creatine were present in both the hepatopancreas and the haemolymph. Phosphorylethanolamine, phosphocholine and fumarate were present only in the hepatopancreas and isoleucine only in the muscle. Statistical analysis showed that the percentage of weight gain was statistically higher in polyphenol‐enriched diet groups compared to the control and that polyphenols had a stimulating effect on the general metabolism. No stress‐related metabolites were higher in crayfish fed with polyphenol‐enriched diet. Conversely, phosphatidylcholine, cholesterol and DHA, linked to resistance to environmental stress and diseases, were higher compared to the control diet. This study indicates that ¹H‐NMR is a useful tool to study the metabolomics in relation to diet differences.     

Coping with change in predation risk across space and time through complementary behavioral responses

Background

Our picture of behavioral management of risk by prey remains fragmentary. This partly stems from a lack of studies jointly analyzing different behavioral responses developed by prey, such as habitat use and fine-scale behavior, although they are expected to complement each other. We took advantage of a simple system on the Kerguelen archipelago, made of a prey species, European rabbit Oryctolagus cuniculus, a predator, feral cat Felis catus, and a mosaic of closed and open foraging patches, allowing reliable assessment of spatio-temporal change in predation risk. We investigated the way such a change triggered individual prey decisions on where, when and how to perform routine activities.

Results

Rabbit presence and behavior were recorded both day and night in patches with similar foraging characteristics, but contrasted in terms of openness. Cats, individually recognizable, were more active at night and in closed patches, in line with their expected higher hunting success in those conditions. Accordingly, rabbits avoided using closed patches at night and increased their vigilance if they did. Both day and night, rabbits increased their use of closed patches as compared to open patches in windy conditions, thereby probably reducing the thermoregulatory costs expected under such harsh environmental conditions.

Conclusions

Overall, our data map the landscape of fear in this study system and indicate that prey habitat use and vigilance complement each other. Solely focusing on one or the other tactic may lead to erroneous conclusions regarding the way predation risk triggers prey decisions. Finally, future studies should investigate inter-individual variability in the relative use of these different types of complementary behavioral responses to perceived risk, along with the determinants and outcomes of such tactics.

     

Low-Molecular-Weight Fucoidan Induces Endothelial Cell Migration via the PI3K/AKT Pathway and Modulates the Transcription of Genes Involved in Angiogenesis

Low-molecular-weight fucoidan (LMWF) is a sulfated polysaccharide extracted from brown seaweed that presents antithrombotic and pro-angiogenic properties. However, its mechanism of action is not well-characterized. Here, we studied the effects of LMWF on cell signaling and whole genome expression in human umbilical vein endothelial cells and endothelial colony forming cells. We observed that LMWF and vascular endothelial growth factor had synergistic effects on cell signaling, and more interestingly that LMWF by itself, in the absence of other growth factors, was able to trigger the activation of the PI3K/AKT pathway, which plays a crucial role in angiogenesis and vasculogenesis. We also observed that the effects of LMWF on cell migration were PI3K/AKT-dependent and that LMWF modulated the expression of genes involved at different levels of the neovessel formation process, such as cell migration and cytoskeleton organization, cell mobilization and homing. This provides a better understanding of LMWF’s mechanism of action and confirms that it could be an interesting therapeutic approach for vascular repair.     

Modeling phytophagous mirid nymphs in cool-climate vineyards

In temperate regions, mirids may be occasional pests of vineyards where risk-averse growers use control measures. In that context, an optimal sampling scheme would allow determination of risk at minimal cost. A model driven by the accumulation of degree-days (DD) was developed to predict cumulative total captures of mirid nymphs (mainly the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois)) at 5%, 50%, and 95%. The model was based on 23 data sets collected over 4 years in three vineyards in southern Quebec. Lower and optimum temperatures for development were set at 10^o and 32°C, based on experimental results and previous studies. Cumulative total captures at 5%, 50%, and 95% levels occurred, respectively, at 229, 331, and 479 DD for the first generation and 630, 806, and 1000 DD for the second generation. Paired t-tests and the forecasting efficiencies confirmed the reliability of the predictive model. The model indicated that monitoring for mirid nymphs of cool-climate in vineyards should be initiated at 200 and 500 DD calculated from 1 March for the first and second generations, respectively. Using the same lower and optimum temperatures for the development of mirids, another degree-day model was developed to predict grapevine phenological stages in relation to the modified Eichhorn-Lorenz system. Superimposing the mirid model over the grapevine model allows relating mirid total captures to grapevine phenological stages, and thus optimizing resources to monitor mirid populations in cool-climate vineyards.     

Development and characterization of a new set of 164 polymorphic EST‐SSR markers for diversity and breeding studies in rubber tree (Hevea brasiliensis Müll. Arg.)

Despite its economic importance and recent genome release, the need for molecular tools for Hevea brasiliensis is high. In the frame of a disease resistance study, EST sequences were retrieved from public database or generated by sequencing SSH libraries. Sequences were trimmed and microsatellite motifs searched using an ad hoc bioinformatic pipeline, and pairs of primers for the amplification of candidate markers were generated. We found a total of 10 499 unigenes from both sources of sequences, and 673 microsatellites motifs were detected using the default parameters of the pipeline. Two hundred sixty‐four primer pairs were tested and 226 (85.6%) successfully amplified. Out of the amplified candidate markers, 164 exhibited polymorphism. Relationships based on dendrograms using simple matching index and diversity statistics based on EST‐SSRs were compared with Genomic SSRs, showing the potentialities of EST‐derived microsatellites for resistance studies but also for population genetics approaches.     

Specific proteins allow classification of pigs according to sire breed, rearing environment and gender

The present study used a proteomic data set obtained from a 2 × 2 × 2 factorial experiment on longissimus muscle of 24 pigs to identify single or couples of proteins allowing correct classification of pigs according to gender, sire breed, or rearing environment. An actin isoform, a myosin light chain 2 isoform and cytochrome Bc1 allowed each, correct classification of all pigs according to gender. Peroxiredoxin 6 allowed correct classification of 23 pigs according to indoor or outdoor rearing environment, but only if gender was also taken into account. Heat shock protein 73 allowed correct classification of 21 pigs according to sire breed, Duroc or Large White. Results show that proteins may be used as biomarkers for traceability or genetic research. Relationships between treatment factors and intracellular regulatory mechanisms associated with these proteins are discussed.     

Canine visceral leishmaniasis: comparison of in vitro leishmanicidal activity of marbofloxacin, meglumine antimoniate and sodium stibogluconate

The control of canine leishmaniasis largely depends on the success of treatment. Drugs currently available to treat this disease are toxic and partially effective. The curative effect of marbofloxacin, a third-generation fluoroquinolone developed for veterinarian individual treatment, was evaluated in vitro in the presence of Leishmania infantum promastigotes and dog-monocyte-derived macrophages; meglumine antimoniate and sodium stibogluconate were used as comparative treatments. We observed that the killing of Leishmania promastigotes and intracellular amastigotes by marbofloxacin was dose-dependent. We demonstrated that successful treatment of canine infected macrophages for 48 h was possible with 500 microg/ml of marbofloxacin. Leishmanicidal activity acted through a TNF-alpha and nitric oxide pathway and correlated with the generation of nitric oxide (NO(2)) production by monocytes derived macrophages from infected (23+/-5 microM) or healthy (21+/-6 microM) dogs, in comparison with NO(2) concentration in infected/non-treated macrophages (< 3 microM, P<0.01). This significant induced parasiticidal effect correlated with extensive elimination of amastigotes by macrophages derived from infected (11+/-5) and healthy dogs (6+/-2), when compared to infected/non-treated macrophages (530+/-105 and 472+/-86 amastigotes, respectively, P< 0.01). Marbofloxacin was shown to be non-toxic at 500 microg/ml in vitro and no cell apoptosis was observed. The molecule was able to induce a parasitic process after significant elimination of amastigotes in leishmania-infected dog macrophages. We propose that marbofloxacin, compared to standard chemotherapeutic agents (meglumine antimoniate and sodium stibogluconate), could be an effective and pragmatic oral route alternative to treat canine leishmaniasis.     

Morphological and molecular characteristics of Echinococcus multilocularis and Echinococcus granulosus mixed infection in a dog from Xinjiang, China

The Xinjiang plateau of western China has been shown to have a high prevalence for human cystic echinococcosis (CE) caused by Echinococcus granulosus, and human alveolar echinococcosis (AE) caused by Echinococcus multilocularis. The domestic dog is suspected to be the primary definitive host for the transmission of both E. granulosus and E. multilocularis to humans in this locality. Seventeen of 30 stray dogs from Hejing County of Xinjiang were found positive for E. granulosus post mortem, and one double infection was suspected. Worm samples were collected, dyed by carmine, and observed microscopically. Carmine staining examination clearly revealed the differences in number of proglottids and appearance of uterine branches and lateral genital pore for those two species of Echinococcus. Furthermore, gene target DNA fragments were amplified for formal identification of the two parasite species, based on 12s rRNA mitochondrial gene. The PCR products were purified and sequenced. Compared with NCBI GenBank, the DNA sequences demonstrated 100% identity with E. granulosus (sheep strain, G1 genotype) and E. multilocularis.