Volatile organic compounds released by blowfly larvae and pupae: new perspectives in forensic entomology.

To evaluate postmortem intervals (PMIs), one should take into account the determined age of necrophagous flies present on the cadaver. However, PMI determination needs further improvement, and rapid and accurate approaches have therefore to be developed. While previous studies have focussed on insect cuticular hydrocarbons, here we explore the volatile profile released by larvae and pupae of Calliphora vicina Robineau-Desvoidy (Diptera: Calliphoridae). We monitored changes in volatile compounds daily, by headspace solid-phase microextraction, followed by gas chromatography-mass spectrometry. Branched and unbranched hydrocarbons, alcohols, esters and acids were identified, and the volatile profile was shown to vary, in both composition and quantity, with the age of the larva/pupa under investigation. We concluded, based on the analysis of the released volatile organic compounds, that it is possible to increase the accuracy of the estimated PMI, through improved estimation of the age of blowflies present on the cadaver.

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.

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.

Analysis of volatile compounds produced by 2 strains of Lactococcus lactis isolated from leben (Tunisian fermented milk) using solid-phase microextraction-gas chromatography.

The volatile compounds that characterize Leben during fermentation with 2 Lactococcus lactis strains (SLT6 and SLT10) in flasks, in a 100-L fermentor, and during storage at 4 degrees C, were investigated and compared to those from commercial Leben. Volatile compounds from Leben were concentrated by a Carboxen-PDMS fiber and analyzed by GC-MS. These compounds include acids, alcohols, aldehydes, ketones, sulfur compounds, and hydrocarbons. Commercial Leben presented a poor volatile profile compared to the laboratory-made Leben. The mixed culture of 2 Lactococcus lactis strains resulted in higher volatile compound formation than the single strain culture. The GC volatile profiles of Leben produced in flask and in the 100-L fermentor were similar. Changes in volatile compounds were observed during storage at 4 degrees C. The effect of culture conditions on production of volatiles by SLT6 strain was studied. Aeration (0.1 mL/min) and agitation enhanced the production of diacetyl, acetoin, 3-methylbutanal, and 3-methylbutanol. Fermentation at pH 5 had no effect on volatile production.

What makes Episyrphus balteatus (Diptera: Syrphidae) oviposit on aphid infested tomato plants?

Under attack by insect pests, many plant species change their volatile chemical emissions to attract natural enemies. Most of the tomato (Lycopersicon sp., Solanaceae) varieties are subjected to infestation by molluscs and insects, including the generalist aphid Myzus persicae Sulzer (Homoptera, Aphididae). Episyrphus balteatus De Geer (Diptera: Syrphidae) is a generalist aphid predator that was here observed to lay eggs on M. persicae infested tomato but not on non-infested plants. In order to identify the volatile chemicals that guide the foraging and oviposition behaviour of E. balteatus, we collected and identified volatiles released in the headspace of both aphid infested and uninfested tomato plants by SPME-GC-MS. The identified chemicals were subsequently tested by electroantennography (EAG) on E. balteatus. Monoterpenes and sesquiterpenes were identified, the main volatile chemicals being beta-phellandrene, 2-carene, alpha-phellandrene, 3-carene and o-pinene. Electrical depolarizations were observed for each tested monoterpene, with optimal responses ranging from -0.2 to -0.8 mV. Episyrphus balteatus antennae showed dose-response relationships towards all the active chemicals. (E)-beta-farnesene, the main component of the aphid alarm pheromone, was the only active sesquiterpene, and is presumed to act as an oviposition stimulus for E. balteotus.

Modelling migration from high-density polyethylene containers into concentrated solutions used as food flavourings.

Migration from high-density polyethylene into different liquids (hexane, ethanol, lemon terpenes and their emulsions) was modelled using the response surfaces method. Polynomial equations (z = A + Bx + Cy + Dx2 + Ey2 + Fxy) were established and parameters determined for each compound. Correlation coefficients were generally > 90%. Analysis indicated that 2,4-bis(1,1-dimethylethyl)phenol, probably due to the degradation of the antioxidant additive tris(2,4-bis(1,1-dimethylethyl)phenyl)phosphite, migrated into each liquid tested, whatever the temperature. Oligomers (10-30 carbons) terminated by a vinyl group were also detected.

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.

Analysis of 5-vinyl-1,3-oxazolidine-2-thione in complex matrices at ppb level.

5-Vinyl-1,3-oxazolidine-2-thione (5-VOT) is a goitrogenic compound released by enzymatic degradation of progoitrin, the most important glucosinolate occurring in rapeseed meal. This paper describes an analytical method for determining the 5-VOT in complex matrices. The method proposed by Quinsac et al. [J. Assoc. Off. Anal. Chem., 75(3) (1992) 529] has been improved by modification of the extraction conditions and the purification steps. The extraction of 5-VOT is performed with hot acid buffer. The first purification step is achieved by solid-phase chromatography (C(18)). The second purification step of 5-VOT is carried out by complexation with phenyl mercury acetate in cyclohexane and, afterwards, by decomplexation using an aqueous solution of sodium thiosulfate. These reactions move 5-VOT from an aqueous to an organic medium, and then back again to the aqueous phase. This procedure ensures a high purification efficiency. The precise quantification of 5-VOT is completed in 12 min by reverse-phase liquid chromatography (C(18)), using an isocratic elution with an ultraviolet detector and with synthetic 4,4-dimethyl-1,3-oxazolidine-2-thione as internal standard. Using this modified method, 5-VOT can be determined in different matrices such as rapeseed meal, animal diets, muscle, several organs (thyroid, liver, kidney and lung) and biological fluids (plasma and milk). The quantification limit of 5-VOT in the sample is of 1 ppb (1 mug kg(-1)), the recovery rate of 5-VOT is about 90% and the repeatability is over 97%.

Measurement of glucosinolates in rapeseeds.

Several ring tests have been organized by the International Organisation for Standardization (ISO) and the Measurement and Testing Programme (BCR) for improving glucosinolate methods. Finally, HPLC of desulphoglucosinolates is recommended by ISO, the European Committee for Standardization (CEN), and the European Commission (EC) as the official method. X-ray fluorescence has also become particularly common for fast analysis. After checking stability of intact glucosinolates in whole rapeseeds, the BCR has three reference materials (CRM 366, CRM 190, CRM 367) available, certified for their total glucosinolate and sulphur contents. Currently, the Measurement and Testing Programme is supporting a research project concerning the stability of desulphoglucosinolates and intact glucosinolates extracts. At the present time, best results are observed with lyophilized extracts sealed in brown glass ampoules and stored at -18 degrees C.

Characterization of Three Labdane Diterpenes from Aframomum alboviolaceum.

Two labdane diterpenes were isolated from the seeds and the rhizomes of AFRAMOMUM ALBOVIOLACEUM (Ridley) K. Schum (Zingiberaceae) and identified by GC-MS, (1)H-, and (13)C-NMR as ( E)-labda-8(17),12-diene-15,16-dial ( 1) and ( E)beta,17-epoxy-labd-12-ene-15,16-dial ( 2). A third minor compound could be the methyl ( E)-14xi,15-epoxylabd-8(17),12-dien-16-oate. The simultaneous occurrence of these three molecules has been mentioned only in one other species of the same genus, AFRAMOMUM DANIELLII (1).

Biological activity of the shrubBoscia senegalensis (PERS.) LAM. ex Poir. (Capparaceae) on stored grain insects.

Biological activity of leaves, fruits and extract of the African shrubBoscia senegalensis (PERS.) LAM. ex Poir. was evaluated against five stored-grain insects. When added to cowpeas at 2-4% (w/w), fresh ground fruits and leaves caused 80-100% mortality inCallosobruchus maculants (F.) adults and significantly reduced both emergence and damage of the F1 progeny. Acetone fruit extract exhibited a potent fumigant effect onProstephanus truncatus HORN, C.maculatus, andSitotroga cerealella OLIV.; with LT50 values of 3.8, 2.3, and below 1.5 hr, respectively. LC50 determination forB. senegalensis fruits and leaves as well as pure methylisothiocyanate (MITC) onTribolium castaneum HERBST,Sitophilus zeamais MOTSCH. andC. maculatus showed a differential response of the insects to plant parts or MITC. Quantitative dosage ofBoscia active components and LC50 values obtained for the plant tissues, compared to those of pure molecules, indicate that the biological activity ofB. senegalensis is due to the liberation of MITC from a glucosinolate precursor glucocapparin contained inBoscia fruits and leaves.

[Quantitative determinaton of glycerides by densitometry (author's transl)]. / Dosage de glycérides par densitométrie

After determination of charring conditions and the measurement by densitometry of lipids separated by thin-layer chromatography, a statistical analysis was performed to determine the significance of the results. Calibration curves were studied and response factors calculated for some saturated and unsaturated triglycerides.