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Analysis of extracts of hemolymph obtained from sexually mature alate females of Solenopsis invicta from monogyne colonies resulted in identification of juvenile hormone III (JH III). The average amount of JH III was 0.32+/-0.04 pmol/&mgr;molof hemolymph. Topical application of 0.038 pmol of JH III was sufficient to stimulate alates to shed their wings in the presence of the queen. The time in which alates were induced to dealate decreased linearly with increasing concentrations of JH III from 0.038 to 3.8 pmol. However, higher JH III concentrations deviated from linearity and did not reach dealation times comparable with those that occur after mating flights. Thus, it appears that the mechanism of dealation that occurs when female alates are out of the influence of their queen is different from the one associated with mating flights. Application of 0.42 &mgr;mol of precocene II inhibited dealation of alates in queenless colonies. However, this inhibition was reversed after applying 38 pmol JH III to precocene-treated alates. The sizes of corpora allata (CA) from sexuals treated with JH III did not differ from those of controls. However, the sizes of CA were reduced in alates treated with precocene II. The results indicated that JH was important to dealation.

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An aqueous extract of German cockroach, Blattella germanica (L.), fecal material was evaluated for inducing trail-following behavior in German cockroaches. In arena tests the fecal extract was found to stimulate trail following in 74% of adult male cockroaches. Significantly fewer cockroaches (22%) followed water-treated (control) trails. Residual activity of the fecal extract trails was evaluated by bioassay after the trails had been stored in the refrigerator or in the open air. Although trails stored in the refrigerator showed no decline in activity after 14 d, those stored in the open air declined significantly after 3 d, inducing only 40% of adult male cockroaches to follow the trail. After 7 d the activity of trails stored in the open air was further reduced to 23%. The ability of fecal-extract trails to influence trap catch in monitoring stations was determined by bioassay. Paper trails treated with fecal extract or water were positioned between cockroach harborages and monitoring stations inside 122-cm2 arenas. The presence of the fecal extract-treated trails significantly enhanced trap catch. Mean catch in the traps with fecal extract trails was 28 cockroaches compared with a mean of 11 cockroaches in the control traps. The trap catch ratios of adults to nymphs in the treated and control treatments were not significantly different.

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A 171,000 M(r )polypeptide of Podisus maculiventris (Say) (Heteroptera: Pentatomidae) that constituted 16% of the protein in eggs also constituted up to 25% of the protein in hemolymph of fed females. It was identified as the major or sole apoprotein of vitellogenin. Eggs contained major polypeptides of 171, 106, and 51 kDa. The hemolymph polypeptide was identified with a polypeptide (vitellin) in egg extracts by comparing molecular weights, specificity of occurrence in fed females, and immunological reactivities. Females, starved for 5 days after eclosion to assure complete previtellogenic development, produced vitellogenin within a day after feeding on larval Galleria mellonella, and within 4 days after feeding on an artificial diet. Appearance of vitellogenin preceded ovarian growth by 2-3 days. Two monoclonal antibodies raised against egg proteins of P. maculiventris were selected for their strong reaction against egg extract and female hemolymph and null reaction against male hemolymph. Only one 170-kDa band in egg and hemolymph reacted with the antibodies on denaturing Western blots. These monoclonal antibodies are being used to develop an enzyme-linked immunosorbent assay (ELISA) to quantitate reproductive response of females to diets of differing quality.

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In this paper we examined the conidial attachment of Metarhizium anisopliae on the southern green stink bug, Nezara viridula, using the exuvia and nymphal stage of the host as a substrate for M. anisopliae conidiospores. Initial studies using fluorescein isothiocyanate (FITC)-labeled conidia examined the differential binding of conidia to various sites on the cuticle. Both the topography and the chemistry of the cuticle affected conidial adhesion. Conidia were trapped in areas containing large numbers of setae (e.g., antennal tips, apical portions of tibia and tarsi). Chemical treatments to remove the cuticle proteins did not affect conidial adhesion, but solvent extraction of cuticular lipids significantly reduced the adhesion of M. anisopliae spores. Germination of M. anisopliae conidia attached to N. viridula cuticle was much less than conidia attached to other insect cuticle substrates. After a 24-hr incubation, only 5-20% of the conidia produced detectable germ tubes. The aldehyde (E)-2-decenal, a primary component of the stink bug scent gland, was detected in cuticle extracts and found to be selectively fungistatic to certain entomopathogenic fungi, including M. anisopliae. The hydrocarbon fraction (nC13 and nC21 to nC31 hydrocarbon series) served as a binding substrate for M. anisopliae, but conidia did not degrade these hydrocarbons and did not use them as a carbon source.

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Extracts of 22 fruits were tested for their attractancy toAnastrepha suspensa (Loew), the Caribbean fruit fly. Box-orange, calamondin, carambola, cattley guava, loquat, and Surinam-cherry were about equal in attractiveness to males and females. Nine synthetic chemicals, including four found in box-orange ripe seed, were attractive to females. Five synthetic chemicals, including two in box-orange ripe seed, were attractive to males. Farnesol,α-phellandrene, and 3-carene were highest in attractiveness to both males and females. Females were more attracted than males to 12 synthetic chemicals. These data suggest that host chemicals serve as attractants and that female and male specific attractants and traps could be developed from host kairomone data. These data also suggest that the volatilization of chemicals from water may play an important role in kairomone biology.

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Pheromonal volatiles emitted by irradiated and control 5- to 11-day-old Caribbean fruit flies,Anastrepha suspensa (Loew), were collected on Tenax adsorbent filters and quantified by gas chromatography (GC). The components released were identified by comparison of retention times on GC and by mass spectrometry (MS) with authentic synthetic standards. Pharate adults were irradiated with gamma rays from a(60)Co source at a dose rate of 10.3 Gray (Gy)/min. The total dosages given were 30, 50, 70, and 100 Gy. Pheromone volatiles were collected from adult males when they were between 5 and 11 days of age. The compounds quantified were (Z)-3-nonenol and (Z, Z)-3,6-nonadienol, which eluted from the GC column together and were quantified as one peak,ß-bisabolene, suspensolide, anastrephin, and epianastrephin. Irradiation with 30 Gy did not significantly reduce any pheromonal components, nor did it change the pheromonal blend. In contrast, suspensolide and bisabolene were significantly reduced in flies irradiated with 50 Gy, while the nonenols and epianastrephin were reduced at the 70-Gy dose. Irradiation with the 100-Gy dose reduced all components with the exception of suspensolide.

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Volatiles emitted byAnastrepha suspensa andAnastrepha ludens males were trapped and compared by GC-MS. Both species emitted previously reported nine-carbon alcohols, (Z)-3-nonenol and (Z,Z)-3,6-nonadienol, and epimeric lactones, anastrephin and epianastrephin. A third isomeric lactone, the macrolide suspensolide, which was previously known only fromA. suspensa is reported here for the first time in the volatiles ofA. ludens. We also report that both species released different proportions of the same three sesquiterpene hydrocarbons:α-farnesene,ß-bisabolene, andα-trans-bergamotene. Theß-bisabolene was isolated in sufficient quantity fromA. suspensa volatiles to establish its absolute configuration as the (R)-(+)-enantiomer. Some plausible biosynthetic relationships of farnesol to the isoprenoid lactones and sesquiterpenes identified in this study are discussed. Finally, we report thatA. suspensa produces the monoterpene (Z)-ß-ocimene whileA. ludens volatiles contained limonene.

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Males of the Caribbean fruit fly,Anastrepha suspensa (Loew), typically form leks and attract females by releasing a multicomponent volatile pheromone. Previous reports have identified two nine-carbon alcohols, three lactones, a sesquiterpene, and a monoterpene in the volatiles. The present report is a study of the physiology of male pheromone release and of ecological and social interactions that influence pheromone release by laboratoryreared flies. Volatiles released by males were trapped on Tenax, eluted, separated, and quantitatively measured by gas chromatography. Experiments showed that the volatiles were primarily released from mouth and anus. Sealing the anal opening or the mouth with melted beeswax resulted in up to 40% or greater reduction in most components, and sealing both mouth and anus further reduced release of volatiles, but some volatiles are possibly still released directly from the cuticle. An anal pouch of everted tissue played a major role as a large evaporative surface for release of some of the volatile components. Male flies entrained to a 14∶10 light-dark cycle showed a peak release of volatiles at 11-12 hr into the photophase, but smaller quantities of the same volatiles were released over a broad period during the daylight hours. Laboratory-reared males peaked in pheromone release at 7-10 days and production and release continued through 35 days of age. Single males released significantly more of all components measured than did groups of males. The reduction by aggregations of males may be related to lekking behavior in this fruit fly. The pheromone probably serves to attract females to a lek site, but additional parameters are likely to enter into the choice of male made by the arriving female.

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Controlled infestation ofDendroctonus terebrans (Olivier) on bolts of slash pine,Pinus ellionii Engelm. var.elliottii, elicited greater attraction of wild conspecifics than uninfested bolts. Secondary attraction was not apparent, however, to standing slash pines that had received volunteer attacks, when compared with attraction to unattacked but susceptible trees. Hindguts from in-flight or attacking femaleD. terebrans contained frontalin, and those from in-flight or attacking males containedexo-brevicomin.Trans-pinocarveol,cis-verbenol,trans-verbenol, myrtenal, verbenone, myrtenol, and other compounds were produced by both sexes during gallery construction in host trees. Synthetic frontalin, when deployed with a standard host odor mixture of turpentine and ethanol, was very attractive to maleD. terebrans in field-trapping experiments. The addition of eitherexo-brevicomin orendo-brevicomin to the frontalin-turpentine combination negated the attractive effect of frontalin for males.Trans-verbenol, myrtenol, and verbenone had little effect onD. terebrans behavior. Responses of females did not differ among treatments in any of the 11 field experiments.

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A new rapid procedure for preparing soft internal tissues from insects that allows air drying was found to compare favorably with tissues prepared by critical point drying. In the new procedure, tissues were fixed in 1% glutaraldehyde, dehydrated through a graded ethanol series, immersed in hexamethyldisilazane (HMDS) for 5 minutes, and air dried. Tissues prepared by both the HMDS treatment and by critical point drying were coated with gold for scanning electron microscopy. Tissues prepared by the HMDS treatment did not shrink or distort upon air drying and excellent surface detail was preserved. The HMDS treatment required about 5 minutes, whereas the critical point drying procedure required about 1.5 hours.

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