Structural and biochemical evaluation of Ceratitis capitata odorant-binding protein 22 affinity for odorants involved in intersex communication.

In insects, odorant-binding proteins (OBPs) connect the peripheral sensory system to receptors of olfactory organs. Medfly Ceratitis capitata CcapObp22 shows 37% identity and close phylogenetic affinities with Drosophila melanogaster OBP69a/pheromone-binding protein related protein 1. The CcapObp22 gene is transcribed in the antennae and maxillary palps, suggesting an active role in olfaction. Here, we recombinantly produced CcapObp22, obtaining a 13.5 kDa protein capable of binding multiple strongly hydrophobic terpene compounds, including medfly male pheromone components. The highest binding affinity [half maximal effective concentration (EC50) = 0.48 µM] was to (E,E)-α-farnesene, one of the most abundant compounds in the male pheromone blend. This odorant was used in cocrystallization experiments, yielding the structure of CcapOBP22. The monomeric structure shows the typical OBP folding, constituted by six α-helical elements interconnected by three disulphide bridges. A C-terminal seventh α-helix constitutes the wall of a deep, L-shaped hydrophobic cavity. Analysis of the electron density in this cavity suggested trapping of farnesene in the crystal structure, although with partial occupancy. Superposition of the CcapOBP22 structure with related seven-helical OBPs highlights striking similarity in the organization of the C-terminal segment of these proteins. Collectively, our molecular and physiological data on medfly CcapOBP22 suggest its involvement in intersex olfactory communication.

Effect of Male House Mouse Pheromone Components on Behavioral Responses of Mice in Laboratory and Field Experiments.

Urine of male house mice, Mus musculus, is known to have primer pheromone effects on the reproductive physiology of female mice. Urine-mediated releaser pheromone effects that trigger certain behavioral responses are much less understood, and no field studies have investigated whether urine deposits by male or female mice, or synthetic mouse pheromone, increase trap captures of mice. In field experiments, we baited traps with bedding soiled with urine and feces of caged female or male mice, and recorded captures of mice in these and in control traps containing clean bedding. Traps baited with female bedding preferentially captured adult males, whereas traps baited with male bedding preferentially captured juvenile and adult females, indicating the presence of male- and female-specific sex pheromones in soiled bedding. Analyses of headspace volatiles emanating from soiled bedding by gas chromatography/mass spectrometry revealed that 3,4-dehydro-exo-brevicomin (DEB) was seven times more prevalent in male bedding and that 2-sec-butyl-4,5-dihydrothiazole (DHT) was male-specific. In a follow-up field experiment, traps baited with DEB and DHT captured 4 times more female mice than corresponding control traps, thus indicating that DEB and DHT are sex attractant pheromone components of house mouse males. Our study provides impetus to identify the sex attractant pheromone of female mice, and to develop synthetic mouse pheromone as a lure to enhance the efficacy of trapping programs for mouse control.

Unsaturated Cuticular Hydrocarbons Enhance Responses to Sex Pheromone in Spruce Budworm, Choristoneura fumiferana.

The primary sex pheromone components of the female spruce budworm, Choristoneura fumiferana (Clem.) (Lepidoptera: Tortricidae), are (E)- and (Z)-11-tetradecenal, produced in 95:5 ratio. However, male flight responses to calling females in a wind tunnel were faster and maintained longer than responses to any synthetic aldehyde blend. Analyses of cuticular extracts from spruce budworm adults revealed series of n-alkanes and n-monoalkenes with predominantly odd numbers of carbon atoms from C23- C29 in both sexes. (Z,Z,Z)-3,6,9-tricosatriene and (Z,Z,Z)-3,6,9-pentacosatriene were identified only in cuticular extracts from females. Pheromonally naïve males showed wing fanning and circling responses to forewing scales from females but not to scales from males. Males also exhibited the same strong responses to scales excised from pharate females, indicating that the pheromone components are produced by females prior to emergence. (Z)-11-hexadecenal and (Z)-5-tricosene enhanced male responses to the primary sex pheromone aldehydes in wind tunnel bioassays, including higher proportions of in-flight and copulatory responses by males and increased time on the source. Addition of (Z,Z,Z)-3,6,9-tricosatriene to the 95/5 blend of (E)- and (Z)-11-tetradecenal released close-range copulatory responses including abdomen curling on treated septa. We propose that the sex pheromone blend of C. fumiferana is composed of the 95/5 blend of (E)- and (Z)-11-tetradecenal as primary components, with (Z)-11-hexadecenal, (Z)-5-tricosene and (Z,Z,Z)-3,6,9-tricosatriene fulfilling secondary roles in orientation and close-range courtship.

Trapping Spodoptera frugiperda (Lepidoptera: Noctuidae) Moths in Different Crop Habitats in Togo and Ghana.

The economic impact of the invasion of Spodoptera frugiperda (J.E. Smith, Lepidoptera: Noctuidae) into Africa has so far been limited to maize agriculture but could potentially impact many other crops. Trapping based on pheromone lures provides a cost-effective method for detecting this important pest (commonly known as fall armyworm) and will be essential for large-scale monitoring of populations to determine its geographical distribution and migration behavior as the species equilibrates to its new environment. However, the effective use of pheromone trapping requires optimization for a given location. An earlier report demonstrated that two commercial lures (one 3-component and the other 4-component) that were effective for trapping S. frugiperda in maize fields in Togo, Africa. The current study extends these findings to agricultural areas that differ in plant host composition (maize, pasture grasses, rice, and sorghum) in multiple locations in Ghana and Togo. In two seasons, significantly higher numbers of moths were found in maize, and in one season, higher numbers were found in rice than in sorghum and pasture grass systems. The results confirm the effectiveness of pheromone trapping and identify pheromone lures and trapping methods best suited for the different agroecosystems common to West Africa and that are at risk of infestation by S. frugiperda.

Attractiveness and Sexual Competitiveness of Anastrepha obliqua Males (Diptera: Tephritidae) Fed on a Diet Enriched With Providencia rettgeri.

The West Indian fruit fly, Anastrepha obliqua (Macquart), is the second most important tephritid fruit fly in Mexico, infesting mango, hog plum and guava fruits. To control this pest, the Mexican government has implemented the use of the sterile insect technique (SIT), which involves the mass production, sterilization and release of flies. However, the A. obliqua laboratory males used in SIT are selected to a lesser extent by the wild females during competitiveness tests. The objective of this study was to compare the effects of males fed on fruit fly food enriched with Providencia rettgeri to those in males fed on food alone, assessing male mating competitiveness, capture of females using traps baited with males fed with the enriched diet and sex pheromone components. The results indicated that males fed with the diet enriched with P. rettgeri had increased mating competitiveness and captured more females in the field cage tests. However, no difference was observed in the proportion of volatile sex pheromone components identified during the calling of A. obliqua males. The results suggest the value of incorporating bacteria into the mass rearing technique of A. obliqua adults in order to improve the sexual competitiveness of males from the laboratory compared to wild males.