Elevated Carbon Dioxide Concentration Reduces Alarm Signaling in Aphids.

Insects often rely on olfaction to communicate with conspecifics. While the chemical language of insects has been deciphered in recent decades, few studies have assessed how changes in atmospheric greenhouse gas concentrations might impact pheromonal communication in insects. Here, we hypothesize that changes in the concentration of atmospheric carbon dioxide affect the whole dynamics of alarm signaling in aphids, including: (1) the production of the active compound (E)-ß-farnesene (Eßf), (2) emission behavior when under attack, (3) perception by the olfactory apparatus, and (4) the escape response. We reared two strains of the pea aphid, Acyrthosiphon pisum, under ambient and elevated CO2 concentrations over several generations. We found that an increase in CO2 concentration reduced the production (i.e., individual content) and emission (released under predation events) of Eßf. While no difference in Eßf neuronal perception was observed, we found that an increase in CO2 strongly reduced the escape behavior expressed by an aphid colony following exposure to natural doses of alarm pheromone. In conclusion, our results confirm that changes to greenhouse gases impact chemical communication in the pea aphid, and could potentially have a cascade effect on interactions with higher trophic levels.

Chemical Composition and Acaricidal Activity of Thymus algeriensis Essential Oil against Varroa destructor.

The aim of the present study was to determine the chemical composition and evaluate the acaricidal activity of Thymus algeriensis essential oil (TAEO) against Varroa destructor. This ectoparasitic mite is a pest of the honey bee Apis mellifera. The essential oil from the, aerial parts of T. algeriensis, obtained by hydrodistillation, was obtained in a yield of 2.8± 0.2%, w/w. The TAEO was analyzed by GC and GC/MS. Thirty-four compounds were identified, representing 99.3% of the oil. The main constituents were carvacrol (48.4%), γ-terpinene (14.9%), p-cymene (14.7%), and thymol (5.6%). Four lots 'were constituted at the level of an apiary in order to study the dynamics of the Varroa destructor and its host, Apis mellifera. After diagnosis by the.biological method "install of diapers", the lots were treated at different doses of TAEO (0.1, 0.3 and 0.5%). TAEO was sprayed on top of the hives. The results show that TAEO at 0.5% resulted in a decrease in the rate of infestation of Varroa destructor, causing a mortality rate of 32.6% without negative effect on the nesting of the queen. The essential oil of T. algeriensis could be used as a bioacaricidal agent.

Differentiation in putative male sex pheromone components across and within populations of the African butterfly Bicyclus anynana as a potential driver of reproductive isolation.

Sexual traits are often the most divergent characters among closely related species, suggesting an important role of sexual traits in speciation. However, to prove this, we need to show that sexual trait differences accumulate before or during the speciation process, rather than being a consequence of it. Here, we contrast patterns of divergence among putative male sex pheromone (pMSP) composition and the genetic structure inferred from variation in the mitochondrial cytochrome oxidase 1 and nuclear CAD loci in the African butterfly Bicyclus anynana (Butler, 1879) to determine whether the evolution of "pheromonal dialects" occurs before or after the differentiation process. We observed differences in abundance of some shared pMSP components as well as differences in the composition of the pMSP among B. anynana populations. In addition, B. anynana individuals from Kenya displayed differences in the pMSP composition within a single population that appeared not associated with genetic differences. These differences in pMSP composition both between and within B. anynana populations were as large as those found between different Bicyclus species. Our results suggest that "pheromonal dialects" evolved within and among populations of B. anynana and may therefore act as precursors of an ongoing speciation process.

Analytical methods used to quantify isoflavones in cow's milk: a review.

This paper provides an update and comprehensive review of the analytical methods used for quantifying isoflavones and their metabolites in cow's milk. Isoflavones are secondary plant metabolites that are similar to 17 ß-estradiol in chemical structure. They form one of the most common categories of phytoestrogens. Numerous health benefits have been attributed to isoflavones, but many of these compounds are also considered to be endocrine disruptors, with adverse effects on health. These contradictory trends offer an attractive prospect for future research, and therefore, sensitive and reliable analytical methods are required to clarify various issues about isoflavones. For this review, a structured methodology was used to select 26 relevant articles published between 2005 and 2015 from the Scopus and CAB Abstract databases. The review discusses individual steps of the analytical procedures described in these articles, including sample preparation, instrumental analysis and validation. The most commonly used analytical procedure is sample preparation involving liquid-liquid extraction and an enzymatic hydrolysis step followed by liquid chromatography with mass spectrometry analysis. Currently, however, there is no standardized procedure for the sample preparation and analysis of isoflavones in milk.

Plutella xylostella (L.) infestations at varying temperatures induce the emission of specific volatile blends by Arabidopsis thaliana (L.) Heynh.

The effect of combined abiotic and biotic factors on plant volatile organic compound (VOC) emissions is poorly understood. This study evaluated the VOC emissions produced by Arabidopsis thaliana (L.) Col-0 subjected to 3 temperature regimes (17, 22, and 27°C) in the presence and absence of Plutella xylostella larvae over 2 time intervals (0-4 and 4-8 h), in comparison to control plants. The analyses of VOCs emitted by Arabidopsis plants were made by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). It was found that certain volatile groups (e.g., alcohols, ketones, aldehydes, and terpenes) are induced by both single factors (temperature or larval infestation) and combined factors (temperature and larvae interactions), whereas other volatile groups (e.g., isothiocyanates [ITCs] and nitrile) were specific to the experimental conditions. ITCs (mainly 4-methylpentyl isothiocyanate) were emitted from plants subjected to larval infestation at 17 and 27°C after the 2 time intervals. The proportions of sulfides (mainly dimethyl disulfide) and 4-(methylthio) butanenitrile were significantly higher on herbivore-infested plants at 22°C compared to the other treatments. Overall, our findings indicate that changes in all experimental conditions caused significant changes to the VOC emissions of Arabidopsis plants. Therefore, the interaction between temperature and larval feeding may represent an important factor determining the variability of volatile emissions by plants subjected to multiple simultaneous factors.

The scent of inbreeding: a male sex pheromone betrays inbred males.

Inbreeding depression results from mating among genetically related individuals and impairs reproductive success. The decrease in male mating success is usually attributed to an impact on multiple fitness-related traits that reduce the general condition of inbred males. Here, we find that the production of the male sex pheromone is reduced significantly by inbreeding in the butterfly Bicyclus anynana. Other traits indicative of the general condition, including flight performance, are also negatively affected in male butterflies by inbreeding. Yet, we unambiguously show that only the production of male pheromones affects mating success. Thus, this pheromone signal informs females about the inbreeding status of their mating partners. We also identify the specific chemical component (hexadecanal) probably responsible for the decrease in male mating success. Our results advocate giving increased attention to olfactory communication as a major causal factor of mate-choice decisions and sexual selection.

Analysis of the diastereoisomers of alliin by HPLC.

Garlic has been known for its therapeutic effects for centuries and is used worldwide as a functional food. The concentration of the active molecules could be enhanced by a better knowledge of their biosynthesis. The precursor of these compounds, alliin (a sulfur amino-acid) has been obtained by chemical synthesis. However, this synthesis route also leads to a diastereoisomer as co-product. This work describes the development of an analytical method which allows the separation and quantification of the two diastereoisomers in order to determine in which proportion the natural form can be produced. The HPLC method which was optimized and validated by accuracy profile exploits an original stationary phase consisting of porous graphitic carbon (PGC). Furthermore, the developped method was used to separate the diastereoisomers of methiin, another cysteine sulfoxide, and to analyze an aqueous extract of garlic. The ability to quantify the amount of natural alliin is valuable for further work on garlic molecules and their application for health protection.

Acaricidal activities of Santolina africana and Hertia cheirifolia essential oils against the two-spotted spider mite (Tetranychus urticae).

BACKGROUND: Many plant essential oils show a broad spectrum of activity against pests. This study investigated the effects of two essential oils on Tetranychus urticae, one of the most serious pests in the world. RESULTS: The chemical composition of the two oils was characterised by GC-MS. The most abundant component in the Santolina africana (Jord. & Fourr) oil was terpinen-4-ol (54.96%), while thymol (61%) was prevalent in the Hertia cheirifolia (L.) oil. Mortality and fecundity were measured upon treatment with oil concentrations ranging from 0.07 to 6.75 mg L(-1) with a Potter spray tower. Mite mortality increased with oil concentration, with LC(50) values of 2.35 mg L(-1) for S. africana and 3.43 mg L(-1) for H. cheirifolia respectively. For both oils, a reduction in fecundity was observed at concentrations of 0.07, 0.09 and 0.29 mg L(-1) . Artificial blends of constituents of oils were also prepared and tested with individual constituents missing from the mixture. The results showed that the presence of all constituents was necessary to equal the toxicity of the two natural oils. CONCLUSION: S. africana and H. cheirifolia oils can provide valuable acaricide activity with significantly lower LC(50) values. Thus, these oils cause important mortality and reduce the number of eggs laid by females.

Optimisation of a semiochemical slow-release alginate formulation attractive towards Aphidius ervi Haliday parasitoids.

BACKGROUND: Optimisation of alginate formulations is described in order to develop semiochemical (E-ß-farnesene and E-ß-caryophyllene) slow-release devices in biological control approaches by attracting predators and parasitoids of aphids. Various formulation criteria were optimised with respect to semiochemical encapsulation capacity. Moreover, the optimised formulation was characterised by texturometry and confocal microscopy. The slow-release rates of semiochemicals were calculated in laboratory controlled conditions. The attractiveness of semiochemical formulations towards Aphidius ervi was demonstrated by olfactometry. RESULTS: Two major parameters were highlighted in encapsulation optimisation: the type of alginate (Sigma L) and the type of crosslinker ion (Ca(2+)). Other formulation parameters were optimised: ionic strength (0.5 M), Ca(2+) (0.2 M) and alginate (1.5%) concentrations and the maturation time of beads in CaCl(2) solution (48 h). After physical characterisation of beads, semiochemical slow-release measurements showed that alginate formulations were efficient sesquiterpene releasers, with 503 µg of E-ß-farnesene and 1791 µg of E-ß-caryophyllene totally released in 35 days. The efficiency of semiochemical alginate beads as attractants for female parasitoids was demonstrated, with high percentages of attraction for semiochemical odours (88 and 90% for E-ß-farnesene and E-ß-caryophyllene respectively) and significant statistical results. CONCLUSION: Semiochemical alginate beads can be considered as efficient slow-release systems in biological control. These formulations could be very useful to attract aphid parasitoids on crop fields.

Microorganisms from aphid honeydew attract and enhance the efficacy of natural enemies.

Aphids are one of the most serious pests of crops worldwide, causing major yield and economic losses. To control aphids, natural enemies could be an option but their efficacy is sometimes limited by their dispersal in natural environment. Here we report the first isolation of a bacterium from the pea aphid Acyrthosiphon pisum honeydew, Staphylococcus sciuri, which acts as a kairomone enhancing the efficiency of aphid natural enemies. Our findings represent the first case of a host-associated bacterium driving prey location and ovipositional preference for the natural enemy. We show that this bacterium has a key role in tritrophic interactions because it is the direct source of volatiles used to locate prey. Some specific semiochemicals produced by S. sciuri were also identified as significant attractants and ovipositional stimulants. The use of this host-associated bacterium could certainly provide a novel approach to control aphids in field and greenhouse systems.

Validation of a fast gas chromatographic method for the study of semiochemical slow release formulations.

The validation of a fast GC-FID analytical method for the quantitative determination of semiochemical sesquiterpenes (E-beta-farnesene and beta-caryophyllene) to be used in an integrated pest management approach is described. Accuracy profiles using total error as decision criteria for validation were used to verify the overall accuracy of the method results within a well defined range of concentrations and to determine the lowest limit of quantification for each analyte. Furthermore it allowed to select a very simple and reliable regression model for calibration curve for the quantification of both analytes as well as to provide measurement uncertainty without any additional experiments. Finally, this validated method was used for the quantification of semiochemicals in slow release formulations. The goal was to verify the protection efficiency of alginate gel beads formulations against oxidation and degradation of sesquiterpenes. The results showed that the alginate beads are adequate slow release devices which protect the bio-active molecules during at least twenty days.

Fast gas chromatography characterisation of purified semiochemicals from essential oils of Matricaria chamomilla L. (Asteraceae) and Nepeta cataria L. (Lamiaceae).

The chemical composition of Matricaria chamomilla L. and Nepeta cataria L. essential oils was determined by GC-MS on an apolar stationary phase by comparison of the characteristic fragmentation patterns with those of the Wiley 275L database. The GC-MS chromatograms were compared with those obtained by fast GC equipped with a direct resistively heated column (Ultra Fast Module 5% phenyl, 5 mx 0.1 mm, 0.1 microm film thickness). Analytical conditions were optimised to reach a good peak resolution (split ratio=1:100), with analysis time lower than 5 min versus 35-45 min required by conventional GC-MS. The fast chromatographic method was completely validated for the analysis of mono- and sesquiterpene compounds. Essential oils were then fractionated by column chromatography packed with silica gel. Three main fractions with high degree of purity in E-beta-farnesene were isolated from the oil of M. chamomilla. One fraction enriched in (Z,E)-nepetalactone and one enriched in beta-caryophyllene were obtained from the oil of N. cataria. These semiochemical compounds could act as attractants of aphid's predators and parasitoids.

Electrophysiological and behavioral responses of the multicolored Asian lady beetle, Harmonia axyridis pallas, to sesquiterpene semiochemicals.

The role of two volatile sesquiterpenes, (E)-beta-farnesene and (-)-beta-caryophyllene, in the chemical ecology of the multicolored Asian lady beetle, Harmonia axyridis Pallas, was investigated by using both electrophysiological and behavioral techniques. (E)-beta-Farnesene is the major component of the alarm pheromone of most aphid species, which are preyed on by H. axyridis. (-)-beta-Caryophyllene was previously isolated from the headspace volatiles above overwintering and aggregated H. axyridis females. These sesquiterpenes elicited significant electroantennogram (EAG) activity from both H. axyridis male and female antennae. In a four-arm olfactometer, male and female H. axyridis were highly attracted toward (E)-beta-farnesene, whereas only males were attracted to (-)-beta-caryophyllene. In a bioassay technique that used a passively ventilated plastic box, both male and female H. axyridis aggregated in the (-)-beta-caryophyllene-treated side of the box. These results support the potential usefulness of (E)-beta-farnesene and (-)-beta-caryophyllene in push-pull strategies that use H. axyridis as a biological control agent in aphid-infested sites or to control this new urban pest in residential structures.