A GABA Receptor Modulator and Semiochemical Compounds Evidenced Using Volatolomics as Candidate Markers of Chronic Exposure to Fipronil in Apis mellifera.

Among the various "omics" approaches that can be used in toxicology, volatolomics is in full development. A volatolomic study was carried out on soil bacteria to validate the proof of concept, and this approach was implemented in a new model organism: the honeybee Apis mellifera. Emerging bees raised in the laboratory in pain-type cages were used. Volatolomics analysis was performed on cuticles, fat bodies, and adhering tissues (abdomens without the digestive tract), after 14 and 21 days of chronic exposure to 0.5 and 1 µg/L of fipronil, corresponding to sublethal doses. The VOCs analysis was processed using an HS-SPME/GC-MS method. A total of 281 features were extracted and tentatively identified. No significant effect of fipronil on the volatolome could be observed after 14 days of chronic exposure. Mainly after 21 days of exposure, a volatolome deviation appeared. The study of this deviation highlighted 11 VOCs whose signal abundances evolved during the experiment. Interestingly, the volatolomics approach revealed a VOC (2,6-dimethylcyclohexanol) that could act on GABA receptor activity (the fipronil target) and VOCs associated with semiochemical activities (pheromones, repellent agents, and compounds related to the Nasonov gland) leading to a potential impact on bee behavior.

A comparative genomic approach using mouse and fruit fly data to discover genes involved in testis function in hymenopterans with a focus on Nasonia vitripennis.

BACKGROUND: Spermatogenesis appears to be a relatively well-conserved process even among distantly related animal taxa such as invertebrates and vertebrates. Although Hymenopterans share many characteristics with other organisms, their complex haplodiploid reproduction system is still relatively unknown. However, they serve as a complementary insect model to Drosophila for studying functional male fertility. In this study, we used a comparative method combining taxonomic, phenotypic data and gene expression to identify candidate genes that could play a significant role in spermatogenesis in hymenopterans. RESULTS: Of the 546 mouse genes predominantly or exclusively expressed in the mouse testes, 36% had at least one ortholog in the fruit fly. Of these genes, 68% had at least one ortholog in one of the six hymenopteran species we examined. Based on their gene expression profiles in fruit fly testes, 71 of these genes were hypothesized to play a marked role in testis function. Forty-three of these 71 genes had an ortholog in at least one of the six hymenopteran species examined, and their enriched GO terms were related to the G2/M transition or to cilium organization, assembly, or movement. Second, of the 379 genes putatively involved in male fertility in Drosophila, 224 had at least one ortholog in each of the six Hymenoptera species. Finally, we showed that 199 of these genes were expressed in early pupal testis in Nasonia vitripennis; 86 exhibited a high level of expression, and 54 displayed modulated expression during meiosis. CONCLUSIONS: In this study combining phylogenetic and experimental approaches, we highlighted genes that may have a major role in gametogenesis in hymenopterans; an essential prerequisite for further research on functional importance of these genes.

Physiological and biochemical responses to thermal stress vary among genotypes in the parasitic wasp Nasonia vitripennis.

In ectotherm species such as insects, thermal fluctuations represent a major environmental factor driving development, survival and reproduction of individuals. Reproductive traits are particularly sensitive to heat stress that can induce a permanent sterility, or at least hypofertility, of adult males. This study aims to compare physiological and biochemical responses associated to male performances to an exposure of 24 h to moderately high temperature (36 °C) among three inbred lines of N. vitripennis (AsymC, Cor, Oul). Cor males showed very specific metabolic adjustments compared to the two other lines. By contrast, Oul males showed stronger phenotypic adjustment of its life cycle, and produced metabolic water to compensate water loss by heat stress. Finally, AsymC males had probably more difficulties to acclimate at 36 °C, even for a short period, as their adult longevity was significantly reduced. Thus, the ability of developmental plasticity in N. vitripennis males exposed to heat stress appears to be dependent of their genotypes.

Volatolomics in Bacterial Ecotoxicology, A Novel Method for Detecting Signatures of Pesticide Exposure?

Volatile organic compounds (VOC) produced by microorganisms in response to chemical stressor showed recently increasing attention, because of possible environmental applications. In this work, we aimed to bring the first proof of concept that volatolomic (i.e., VOCs analysis) can be used to determine candidate VOC markers of two soil bacteria strains (Pseudomonas fluorescens SG-1 and Bacillus megaterium Mes11) exposure to pesticides. VOC determination was based on solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS). Accordingly, we highlighted a set of bacterial VOCs modulated in each strains according to the nature of the pesticide used. Three out these VOCs were specifically modulated in P. fluorescens SG-1 when exposed with two pyrethroid pesticides (deltamethrine and cypermethrine): 2-hexanone; 1,3-ditertbutylbenzene and malonic acid, hexyl 3-methylbutyl ester. Our results thus suggest the possible existence of generic VOC markers of pyrethroids in this strain. Of particular interest, two out of these three VOCs, the 1,3-ditertbutylbenzene and the malonic acid, hexyl 3-methylbutyl ester were found also in B. megaterium Mes11 when exposed with cypermethrine. This result highlighted the possible existence of interspecific VOC markers of pyrethroid in these two bacteria. Altogether, our work underlined the relevance of volatolomic to detect signatures of pesticides exposure in microorganisms and more generally to microbial ecotoxicology. Based on these first results, considerations of volatolomics for the chemical risk assessment in environment such as soils can be indirectly explored in longer terms.

Uso de megadosis de vitamina C en la cicatrización y desinflamación de heridas quirúrgicas / Use of megadoses of vitamin C in the healing and disinflammation of surgical wounds

Fundamento: En el campo hospitalario, aún es tema de controversia sobre cuándo recomendar el uso de altas dosis de Vitamina C. Objetivo: Determinar la utilidad clínica de la Vitamina C a grandes dosis por vía intravenosa. Metodología: Se revisaron artículos académicos disponibles en MedLine, EMBASE, Cochrane, ScienceDirect, Scopus, Scielo, Pubmed, Medigraphic, Elsevier y Biblioteca Virtual en Salud. Resultados: El uso de megadosis de vitamina C después de un procedimiento quirúrgico presenta efectos favorables en la resolución del proceso inflamatorio a dosis de 3 gramos al día administrados parenteralmente, a su vez, al ser usado como coadyuvante en la cicatrización de heridas a dosis de 4 gramos al día ejerce un efecto beneficioso contra la dehiscencia de heridas. Estas propiedades se explican por la capacidad del ácido ascórbico para disminuir la actividad de las citocinas proinflamatorias, además de su participación en la síntesis del colágeno tipo I, elemento indispensable para una adecuada formación cicatricial. Conclusiones: Varios estudios comprobaron los beneficios del uso de megadosis de la vitamina C después de un procedimiento quirúrgico.

Background: There is still controversy over how to recommend the use of high doses of Vitamin C in treatment of patients in hospitals. Objective: To determine the clinical usefulness of Vitamin C at high doses intravenously. Methodology: Review of academic articles available in MedLine, EMBASE, Cochrane, ScienceDirect, Scopus, Scielo, Pubmed, Medigraphic, Elsevier and Virtual Health. Results: The use of megadoses of vitamin C afer asurgical procedure has favorable effects in the resolution of the inflammatory process at doses of 3 grams per day administered parenterally. Sometimes it is used as a coadjuvant in the wound healing process at doses of 4 grams per day with beneficial effects and against wound dehiscence. These properties are explained by the ability of ascorbic acid to decrease the activity of proinflammatory cytokines, and in addition to its participation in the synthesis of collagen type I, indispensable element for an adequate scar formation. Conclusions: Several studies have shown the benefits of using megadoses of vitamin C after a surgical procedure.

The spatio-temporal partitioning of sperm by males of the prospermatogenic parasitoid Nasonia vitripennis is in line with its gregarious lifestyle.

Male fitness depends on the number of lifetime progeny of their mates and could be constrained by the chance of finding a mate, lifespan and temporal patterns of sperm production and allocation. Here, we used the parasitic wasp Nasonia vitripennis with a two-week lifespan and a gregarious lifestyle, to analyze how the reproductive system is organized to allocate spermatozoa over consecutive matings. Results show that spermatogenesis is synchronized and completed one day before emergence so that males emerge with a full sperm complement. We also found a regulation of spermatozoa transfer between testis and seminal vesicles that allows males to partition small ejaculates over multiple matings. Overall, this study shows that for N. vitripennis, male fertilization potential is determined (1) at the pupal stage, when spermatogenesis takes place to generate a complete life-long stock, (2) on emergence, when transport of spermatozoa from testes to seminal vesicles is initiated and (3) in adulthood, during which spermatozoa are partitioned over successive copulations. Such life history-traits are consistent with the gregarious lifestyle of N. vitripennis.

Seasonal variation in wing size and shape between geographic populations of the malaria vector, Anopheles coluzzii in Burkina Faso (West Africa).

The mosquito, Anopheles coluzzii is a major vector of human malaria in Africa with widespread distribution throughout the continent. The species hence populates a wide range of environments in contrasted ecological settings often exposed to strong seasonal fluctuations. In the dry savannahs of West Africa, this mosquito population dynamics closely follows the pace of surface water availability: the species pullulates during the rainy season and is able to reproduce throughout the dry season in areas where permanent water bodies are available for breeding. The impact of such environmental fluctuation on mosquito development and the phenotypic quality of emerging adults has however not been addressed in details. Here, we examined and compared phenotypic changes in the duration of pre-imaginal development, body dry mass at emergence and wing size, shape and surface area in young adult females An. coluzzii originated from five distinct geographic locations when they are reared in two contrasting conditions mimicking those experienced by mosquitoes during the rainy season (RS) and at the onset of the dry season (ODS) in Burkina Faso (West Africa). Our results demonstrated strong phenotypic plasticity in all traits, with differences in the magnitude and direction of changes between RS and ODS depending upon the geographic origin, hence the genetic background of the mosquito populations. Highest heterogeneity within population was observed in Bama, where large irrigation schemes allow year-round mosquito breeding. Further studies are needed to explore the adaptive value of such phenotypic plasticity and its relevance for local adaptation in An. coluzzii.

Deciphering the metabolic changes associated with diapause syndrome and cold acclimation in the two-spotted spider mite Tetranychus urticae.

Diapause is a common feature in several arthropod species that are subject to unfavorable growing seasons. The range of environmental cues that trigger the onset and termination of diapause, in addition to associated hormonal, biochemical, and molecular changes, have been studied extensively in recent years; however, such information is only available for a few insect species. Diapause and cold hardening usually occur together in overwintering arthropods, and can be characterized by recording changes to the wealth of molecules present in the tissue, hemolymph, or whole body of organisms. Recent technological advances, such as high throughput screening and quantification of metabolites via chromatographic analyses, are able to identify such molecules. In the present work, we examined the survival ability of diapausing and non-diapausing females of the two-spotted spider mite, Tetranychus urticae, in the presence (0 or 5°C) or absence of cold acclimation. Furthermore, we examined the metabolic fingerprints of these specimens via gas chromatography-mass spectrophotometry (GC-MS). Partial Least Square Discriminant Analysis (PLS-DA) of metabolites revealed that major metabolic variations were related to diapause, indicating in a clear cut-off between diapausing and non-diapausing females, regardless of acclimation state. Signs of metabolic depression were evident in diapausing females, with most amino acids and TCA cycle intermediates being significantly reduced. Out of the 40 accurately quantified metabolites, seven metabolites remained elevated or were accumulated in diapausing mites, i.e. cadaverine, gluconolactone, glucose, inositol, maltose, mannitol and sorbitol. The capacity to accumulate winter polyols during cold-acclimation was restricted to diapausing females. We conclude that the induction of increased cold hardiness in this species is associated with the diapause syndrome, rather than being a direct effect of low temperature. Our results provide novel information about biochemical events related to the cold hardening process in the two-spotted spider mite.

Effects of diet and salinity on the survival, egg laying and metabolic fingerprints of the ground-dwelling spider Arctosa fulvolineata (Araneae, Lycosidae).

Soil salinity and the salinity of trophic resources may alter the osmoregulatory processes of arthropod, challenging the smooth regulation of body water, and, ultimately, survival. The intra and extracellular build-up of osmolytes represent a common strategy to attenuate acute hyperosmotic stress in several arthropod species. In the present study, we aimed to determine the impact of substrate and trophic resource salinities on salt tolerance in the female wolf spider, Arctosa fulvolineata, which is considered a specialist salt marsh species. We evaluated adult female survival and egg laying, and quantified the osmo-induced accumulation of compatible solutes (GC-MS). Three concentrations of substrate salinity were tested (0‰, 35‰ and 70‰) under three trophic conditions (starved spiders, spiders fed with salt prey [intertidal amphipods] and spiders fed with unsalted prey [freshwater amphipods]). We found no support for diet preferences in female A. fulvolineata, which exhibited similar predation rates on freshwater and marine amphipods. Survival and egg-laying were significantly impaired when female A. fulvolineata were exposed to hypersaline conditions for 12 days. Our results showed an increase in the level of several compatible solutes when spiders were exposed for 12 days to saline conditions. For instance, α-alanine, ß-alanine, arginine, asparagine, aspartate, homoserine, glutamine, glycine, proline and serine levels were 4-10 times higher under hypersaline conditions. The osmo-induced accumulation of amino acids may increase the osmolality of body fluids, thus enhancing the smooth regulation of body fluids and survival ability of wolf spider under extreme saline conditions.