About lipid metabolism in Hermetia illucens (L. 1758): on the origin of fatty acids in prepupae.

Although increasingly targeted in animal nutrition, black soldier fly larvae or prepupae (BSF, Hermetia illucens L. 1758) require the characterization and modulation of their fatty acid profile to become fully integrated within the feed sector. This improvement will only be possible by the understanding of underlaying biochemical pathways of fatty acid synthesis in BSF. In this study, we hypothesized a labelling of de novo synthesized fatty acids in BSF by the incorporation of deuterated water (D2O) in their feed. Three batches of fifty larvae were reared on two diets with different polyunsaturated fatty acid profiles moistened with 40% of H2O or D2O: chicken feed or 40% of chicken feed and 60% of flax cake. Although the occurrence of D2O in insect feed increased the larval development time and decreased prepupal weight, it was possible to track the biosynthesis of fatty acids through deuterium labelling. Some fatty acids (decanoic, lauric or myristic acid) were exclusively present in their deuterated form while others (palmitic, palmitoleic or oleic acid) were found in two forms (deuterated or not) indicating that BSF can partially produce these fatty acids via biosynthesis pathways and not only by bioaccumulation from the diet. These results suggest the importance of carbohydrates as a source of acetyl-CoA in the constitution of the BSF fatty acid profile but also the potential importance of specific enzymes (e.g. thioesterase II or Δ12 fat2 desaturase) in BSF fatty acid metabolism. Finally, nearly no deuterated polyunsaturated fatty acids were found in BSF fed with deuterium confirming that BSF is not able to produce these types of fatty acids. Despite the high levels of linolenic acid in flax-enriched diets, BSF will simply bioaccumulate around 13% of this fatty acid and will metabolize approximately two-thirds of it into saturated fatty acids as lauric or myristic acid.

Male flowers of Aconitum compensate for toxic pollen with increased floral signals and rewards for pollinators.

Many plants require animal pollinators for successful reproduction; these plants provide pollinator resources in pollen and nectar (rewards) and attract pollinators by specific cues (signals). In a seeming contradiction, some plants produce toxins such as alkaloids in their pollen and nectar, protecting their resources from ineffective pollinators. We investigated signals and rewards in the toxic, protandrous bee-pollinated plant Aconitum napellus, hypothesizing that male-phase flower reproductive success is pollinator-limited, which should favour higher levels of signals (odours) and rewards (nectar and pollen) compared with female-phase flowers. Furthermore, we expected insect visitors to forage only for nectar, due to the toxicity of pollen. We demonstrated that male-phase flowers emitted more volatile molecules and produced higher volumes of nectar than female-phase flowers. Alkaloids in pollen functioned as chemical defences, and were more diverse and more concentrated compared to the alkaloids in nectar. Visitors actively collected little pollen for larval food but consumed more of the less-toxic nectar. Toxic pollen remaining on the bee bodies promoted pollen transfer efficiency, facilitating pollination.

Proteomic analysis of Arabidopsis thaliana (L.) Heynh responses to a generalist sucking pest (Myzus persicae Sulzer).

Herbivorous insects can cause severe cellular changes to plant foliage following infestations, depending on feeding behaviour. Here, a proteomic study was conducted to investigate the influence of green peach aphid (Myzus persicae Sulzer) as a polyphagous pest on the defence response of Arabidopsis thaliana (L.) Heynh after aphid colony establishment on the host plant (3 days). Analysis of about 574 protein spots on 2-DE gels revealed 31 differentially expressed protein spots. Twenty out of these 31 differential proteins were selected for analysis by mass spectrometry. In 12 of the 20 analysed spots, we identified seven and nine proteins using MALDI-TOF-MS and LC-ESI-MS/MS, respectively. Of the analysed spots, 25% contain two proteins. Different metabolic pathways were modulated in Arabidopsis leaves according to aphid feeding: most corresponded to carbohydrate, amino acid and energy metabolism, photosynthesis, defence response and translation. This paper has established a survey of early alterations induced in the proteome of Arabidopsis by M. persicae aphids. It provides valuable insights into the complex responses of plants to biological stress, particularly for herbivorous insects with sucking feeding behaviour.

USE OF MICRO-DAMS IN POTATO FURROWS TO REDUCE EROSION AND RUNOFF AND MINIMISE SURFACE WATER CONTAMINATION THROUGH PESTICIDES.

The use of micro-dams in potato furrows is an interesting technology to reduce erosion and runoff in hilly areas. These phenomena are major sources of surface water contamination by nutrients and plant protection products (Gillijns et al., 2005). In 2011 Bayer CropScience set up a trial in collaboration with the Walloon Agricultural Research Centre (CRA-W) and ULg-Gembloux Agro-Bio Tech in Huldenberg (Belgium) to demonstrate this technique in potatoes. Micro-dams create barriers between furrows in order to encourage rainwater to infiltrate in the soil rather than to run off. The results from the trial over this year confirm that the application of micro-dams is effective in reducing erosion and runoff significantly. The total loss of plant protection products (PPP) to surface water is dramatically reduced and also strongly depends on the physic-chemical characteristics of the active ingredients. In addition, the technique tends to produce a higher yield of potato tubers as an effect of an optimised utilisation of the available rainwater and nutrients.

Effect of a fungal infection on the profile of volatile organic compounds emitted by plant roots.

It is known since few years that the aerial and underground parts of the plants emit volatile organic compounds (VOCs) that can interact with other organisms of the environment. They are involved in the attraction of seed dispersers and pollinators, the repellence of enemies via direct or indirect mechanisms and the induction of defence systems in other parts of the same plant or in other plants in the vicinity (Dudareva et al., 2006). It has been shown previously that the VOCs spectrum emitted by plants hardly depends on their physiological state (Kant et al., 2009). However those phenomenons were poorly studied at the edaphic level. Thus, the Rhizovol project, a multidisciplinary project in Gembloux Agro-Bio Tech was set up to study the emissions of VOCs by plant roots and their interactions with other organisms of the rhizosphere. As a partner of this project, the Plant Pathology Unit of Gembloux Agro-Bio Tech chose to study the effect of a fungal infection on the profile of VOCs emitted by plant roots, based on three model organisms, barley (Hordeum vulgare L.), since it is a major crop in Belgium that can suffer a large range of aggressions, and two pathogenic fungi, Cochliobolus sativus and Fusarium culmorum, responsible for root and foot rots and seedling blight on cereals (Wiese, 1977). Later in the development, C. sativus produces elongate brown-black lesions (spot blotch) and F. culmorum induces head blight and produces mycotoxins that make the grain unsuitable for consumption (Nielsen et al., 2011). The objective of this work was to identify the VOCs emitted during the dual interactions between barley roots and a pathogenic fungus. The study was performed in two steps; first, the independent analyses of the VOCs emitted by each of the partners (C. sativus, F. culmorum and healthy barley roots), then the analyses of the VOCs spectrum emitted during dual interactions.

Chemical composition and acaricidal properties of Deverra scoparia essential oil (Araliales: Apiaceae) and blends of its major constituents against Tetranychus urticae (Acari: Tetranychidae).

The essential oil of Deverra scoparia Coss. & Durieu was investigated for its acaricidal activity against the worldwide pest twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae). The essential oil was analyzed by fast gas chromatography (GC) and GC-mass spectrometry. The activities of its individual and blended constituents were determined. Our study showed that female mortality increased with increasing D. scoparia oil concentrations, with LD50 and LD90 values at 1.79 and 3.2 mg liter(-1), respectively. A reduction in fecundity had already been observed for concentrations of 0.064, 0.08, and 0.26 mg liter(-1) D. scoparia essential oil. Ten major components, comprising 98.52% of the total weight, were identified; a-pinene was the most abundant constituent (31.95%) followed by sabinene (17.24%) and delta3-carene (16.85%). The 10 major constituents of D. scoparia oil were individually tested against T. urticae females. The most potent toxicity was found with alpha-pinene, delta3-carene, and terpinen-4-ol. The presence of all constituents together in the artificial mixture caused a significant decrease in the number of eggs laid by females, at 0.26 mg liter(-1) (11 eggs), compared with the control (50 eggs). The toxicity of blends of selected constituents indicated that the presence of all constituents was necessary to reproduce the toxicity level of the natural oil.

Does Tribolium brevicornis cuticular chemistry deter cannibalism and predation of pupae?

The cuticular hydrocarbons of insects are species-specific and often function as semiochemicals. The activity of Tribolium brevicornis cuticular hydrocarbons as feeding deterrents that ostensibly function to prevent pupal cannibalism and predation was evaluated. The cuticular hydrocarbons of T. brevicornis pupae were characterized and flour disk bioassays conducted with individual and combined extract components incorporated into artificial diets on which Tribolium adults fed for six days. Feeding by T. brevicornis and T. castaneum on flour disks containing cuticular extracts of T. brevicornis pupae resulted in reduced consumption and weight loss relative to feeding on control flour disks. In both cases, feeding deterrence indices exceeded 80% suggesting that T. brevicornis cuticular hydrocarbons could function to deter cannibalism and predation of pupae by larvae and adult beetles. Sixteen different cuticular hydrocarbons were identified in T. brevicornis pupal extracts. Eight of the commercially available linear alkanes were tested individually in feeding trials with eight Tribolium species. One compound (C28) significantly reduced the amount of food consumed by three species compared to control disks, whereas the compounds C25, C26, and C27 elicited increased feeding in some species. Four other compounds had no effect on consumption for any species. When four hydrocarbon mixtures were tested for synergistic deterrence on T. brevicornis and T. castaneum, none significantly influenced consumption. Our results indicate that the cuticular chemistry of T. brevicornis pupae could serve to deter predation by conspecific and congeneric beetles.

Study of Thermomyces lanuginosa lipase in the presence of tributyrylglycerol and water.

The Thermomyces lanuginosa lipase has been extensively studied in industrial and biotechnological research because of its potential for triacylglycerol transformation. This protein is known to catalyze both hydrolysis at high water contents and transesterification in quasi-anhydrous conditions. Here, we investigated the Thermomyces lanuginosa lipase structure in solution in the presence of a tributyrin aggregate using 30 ns molecular-dynamics simulations. The water content of the active-site groove was modified between the runs to focus on the protein-water molecule interactions and their implications for protein structure and protein-lipid interactions. The simulations confirmed the high plasticity of the lid fragment and showed that lipid molecules also bind to a secondary pocket beside the lid. Together, these results strongly suggest that the lid plays a role in the anchoring of the protein to the aggregate. The simulations also revealed the existence of a polar channel that connects the active-site groove to the outside solvent. At the inner extremity of this channel, a tyrosine makes hydrogen bonds with residues interacting with the catalytic triad. This system could function as a pipe (polar channel) controlled by a valve (the tyrosine) that could regulate the water content of the active site.

Cadaveric volatile organic compounds released by decaying pig carcasses (Sus domesticus L.) in different biotopes.

Forensic entomology uses pig carcasses to surrogate human decomposition and to investigate the entomofaunal colonization. Insects communicate with their environment through the use of chemical mediators, which in the case of necrophagous insects, may consist in the cadaveric volatile organic compounds (VOCs) released by the corpse under decomposition. Previous studies have focused on cadaveric VOCs released from human corpses. Nevertheless, studies on human corpses are restricted for many reasons, including ethics. Forensic entomologists use pig as animal model but very few information are available about the decompositional VOCs released by a decaying pig carcass. We here tested a passive sampling technique, the Radiello diffusive sampler, to monitor the cadaveric VOCs released by decomposing pig carcasses in three biotopes (crop field, forest, urban site). A total of 104 chemical compounds, exclusively produced by the decompositional process, were identified by thermal desorption interfaced with gas chromatography and mass spectrometry (TDS-GC-MS). Ninety, 85 and 57 cadaveric VOCs were identified on pig carcasses laying on the agricultural site, the forest biotope and in the urban site, respectively. The main cadaveric VOCs are acids, cyclic hydrocarbons, oxygenated compounds, sulfur and nitrogen compounds. A better knowledge of the smell of death and their volatile constituents may have many applications in forensic sciences.

Enzymatically prepared n-alkyl esters of glucuronic acid: the effect of freeze-drying conditions and hydrophobic chain length on thermal behavior.

In this work, some of the physicochemical properties of enzymatically prepared n-alkyl esters of glucuronic acid are presented. Two questions are addressed. The first concerns the influence of post-purification freeze-drying conditions on octyl glucuronate thermotropic behavior. Depending on the amount of water added before freeze-drying, the alpha/beta anomeric ratio determined by (1)H NMR is affected and differences are observed in DSC thermograms probably due to polymorphism. The second question concerns the effect of hydrophobic chain length on the thermal behavior. An increase of both transition temperature and transition enthalpy is observed by increasing the number of carbon atoms in the alkyl chain (C8

Electrophysiological and behavioral activity of secondary metabolites in the confused flour beetle, Tribolium confusum.

Several previous studies have addressed pheromone communication in various flour beetles (Coleoptera: Tenebrionidae), including the confused flour beetle, Tribolium confusum (du Val). Different stereoisomers of 4,8-dimethyldecanal (DMD) were reported as the only components of an aggregation pheromone, but the behavioral activity of DMD is low. In the present study, additional previously reported secondary metabolites (benzoquinones and hydrocarbons) were tested for electrophysiological activity (EAG) with both sexes of T. confusum. Two benzoquinones and three monoenic hydrocarbons elicited significant EAG activity from both male and female antennae. There was an elevated male EAG response (vs. the females) to two out of the three hydrocarbons and for both quinones. The EAG-active compounds were subsequently investigated for behavioral activity in a walking bioassay. Benzoquinones are considered toxic and have been assigned a function as alarm substances in flour beetles, but we found that methyl-1, 4-benzoquinone in intermediate concentrations was attractive to both male and female beetles and could therefore act as an aggregation pheromone component. Males were also attracted to ethyl-1,4-benzoquinone. The corresponding hydroquinones, presumed precursors of the benzoquinones, did not elicit any electrophysiological response and were not tested for behavioral activity. The unsaturated hydrocarbons (1-tetradecene, 1-pentadecene, and 1-hexadecene) elicited significant EAG responses from both male and female antennae and were also attractive in the behavioral assay. Our results show that several beetle-produced compounds, in addition to 4,8-dimethyldecanal, may be part of a complex pheromone system in flour beetles and play a role in mediating aggregation in T. confusum.

Estimating fatty acid content in cow milk using mid-infrared spectrometry.

Interest in the fatty acid composition of dairy products is increasing; however, the measurement of fatty acids requires using gas-liquid chromatography. Although this method is suitable, it involves a time-consuming procedure, expensive reagents, and qualified staff. By comparison, the mid-infrared (MIR) spectrometry method could be a good alternative for assessing the fatty acid profile of dairy products. The objective of this study was to explore the calibration of MIR spectrometry for estimating fatty acid concentrations in milk and milk fat. Estimated concentrations in milk fat were less reliable than those for the same fatty acids in milk. Results also showed that when the fatty acid concentrations in milk increased, the efficiency of the infrared analysis method in predicting these values simultaneously increased. Selected prediction equations must have a high cross-validation coefficient of determination, a high ratio of standard error of cross-validation to standard deviation, and good repeatability of chromatographic data. Results from this study showed that the calibration equations predicting 12:0, 14:0, 16:0, 16:1cis-9, 18:1, and saturated and monounsaturated fatty acids in milk could be used. Thus, with its potential for use in regular milk recording, this infrared analysis method offers the possibility of assessing and improving the quality of milk produced. Indeed, it enables the fatty acid composition in milk to be estimated for each cow and the estimates to be used as indicator traits to determine the genetic values of underlying fatty acid concentrations. The knowledge of these genetic values would open up opportunities for animal selection aimed at improving the nutritional quality of cow milk.

Nematode infection and reproduction in transgenic and mutant Arabidopsis and tobacco with an altered phenylpropanoid metabolism.

Transgenic and mutant Arabidopsis and tobacco plants with altered phenylpropanoid metabolism were infected with the plant parasitic root knot nematode Meloidogyne incognita to assess the effect of the transgene or mutation on nematode infection and reproduction. Modifications in the lignin biosynthetic pathway which alter lignin composition in roots affected reproduction. In Arabidopsis with increased levels of syringyl lignin, reproduction was lower than in wild-type plants, while in tobacco with reduced levels of syringyl lignin, life cycle progression was stimulated. Overexpression of a MYB transcription factor of phenylpropanoid metabolism in tobacco significantly stimulated reproduction of M. incognita, while overexpression of L-phenylalanine ammonia-lyase had no effect. Arabidopsis transparent testa mutants with deficiencies in flavonoid pathway enzymes did not affect reproduction of M. incognita in the present infection tests.

New model for performance prediction in fixed-bed reactors based on the approach of the unused bed zone.

In the present study a practical model useful for designers dealing with pilot scale reactors for pollutant control has been developed. Investigations were undertaken for a single-component lead study on New Zealand clinoptilolite at the temperature of 25+/-1 degrees C. Fifteen runs under different operating conditions such as particle characteristics (expressed as d*(p)/D), column geometry (as l*/D) and flow speed (as R*e) have been performed. The results were interpreted by a response surface method (RSM) from which an equation giving the unused bed zone (UBZ) was obtained. From those dimensionless parameters, a sequence of contour plots was drawn, making it easy for a designer to choose optimum design parameters while controlling the operation performance. The flow rate (R*e) range was established over the laminar flow conditions from 1 to 8, (l/D*) was extended from 7 to 20 and (d(p)/D*) varied from 3% to 10%. Optimization of performance of the reactor as UBZ could vary from 10% to 40% of the material operating capacity.

Potential use of essential oils from Cameroon applied as fumigant or contact insecticides against Sitophilus zeamais Motsch. (Coleoptera: Curculionidae).

Essential oils from seven plants species currently found in Cameroon were extracted by steam distillation and tested for their insecticidal activities against Sitophilus zeamats Motsch. Responses varied with the test applied and the plant species. For the contact toxicity, the acetone was used in order to dilute the pure essential oil. Formulations of 1% of essential oils of Xylopia aethiopica and Ocimum gratissimum were the most toxic and led to 96 and 98% of mortality respectively after 24 h. There was no death in control (0% of mortality). Fumigation test were done by applying 300 microl of pure essential oils in the bottom of closed 800 ml glass containers. In that case, the essential oil of Hyptis spicigera was the most toxic (fumigant) after 48 hours of fumigation followed by Annona seregalensis and Xylopia aethiopica 96 and 95% of mortality respectively. Some of the essential oils of plant species tested are promising for pest control in farmer granaries. However, further investigations are to be done on formulation and side effects.

The prevalence of mycotoxins in Kashin-Beck disease.

Mycotoxins are naturally occurring toxic chemical compounds produced by fungi infesting agricultural crops both during their growth and storage. Such secondary metabolites, when ingested, can produce toxic syndromes in humans. As it has been suggested that mycotoxins might be involved in the development of Kashin-Beck disease (KBD), we undertook a survey of barley grains of KBD-affected families and non-affected families in that country. We found, by thin layer chromatography, a hitherto unknown metabolite of Alternaria sp. This was especially common on the barley grains of KBD-affected families.

Effects of allelochemicals from first (brassicaceae) and second (Myzus persicae and Brevicoryne brassicae) trophic levels on Adalia bipunctata.

Three Brassicaceae species, Brassica napus (low glucosinolate content), Brassica nigra (including sinigrin), and Sinapis alba (including sinalbin) were used as host plants for two aphid species: the generalist Myzus persicae and the specialist Brevicoryne brassicae. Each combination of aphid species and prey host plant was used to feed the polyphagous ladybird beetle, Adalia bipunctata. Experiments with Brassicaceae species including different amounts and kinds of glucosinolates (GLS) showed increased ladybird larval mortality at higher GLS concentrations. When reared on plants with higher GLS concentrations, the specialist aphid, B. brassicae, was found to be more toxic than M. persicae. Identification of GLS and related degradation products, mainly isothiocyanates (ITC), was investigated in the first two trophic levels, plant and aphid species, by high-performance liquid chromatography and gas chromatography-mass spectrometry, respectively. While only GLS were detected in M. persicae on each Brassicaceae species, high amounts of ITC were identified in B. brassicae samples (allyl-ITC and benzyl-ITC from B. nigra and S. alba, respectively) from all host plants. Biological effects of allelochemicals from plants on predators through aphid prey are discussed in relation to aphid species to emphasize the role of the crop plant in integrated pest management in terms of biological control efficacy.