Intercaste,intercolony, and temporal variation in cuticular hydrocarbons ofCopotermes formosanus shiraki (Isoptera: Rhinotermitidae)

We characterized the variation in cuticular hydrocarbon mixtures between seven colonies of the Formosan subterranean termite,Coptotermes formosanus Shiraki, from the same population. We report differences between castes, between colonies, and within the population over time to assess seasonality. Colonies ofC. formosanus from Oahu, Hawaii, were sampled for 25 months. Each month, one sample each of 200 workers, 50 soldiers, nymphs, or alates from each colony was subjected to gas chromatography-mass spectrometry (GC-MS) analysis of the cuticular hydrocarbons. We resolved 39 individual peaks and identified 52 individual or isomeric mixtures of hydrocarbons. Onlyn-alkanes and methyl-branched alkanes occur; no olefins were found. Internally branched monomethylalkanes were the most abundant class of hydrocarbons, representing 45% to 50% of the total 9-;11-;13-Methyl-heptacosane accounted for over 30% of the total hydrocarbon for all castes. 2-Methyl- and 3-methylalkanes comprise approximately 30% of the total. Internally branched dimethylalkanes constitute 15% to 20% of the total cuticular hydrocarbon. Only one trimethylalkane, 13,15,17-trimethylnonacosane, was found in small amounts. The hydrocarbon mixtures of all four castes were similar. Quantitative differences in hydrocarbon mixtures among the castes were easily displayed using canonical discriminant analysis. Soldiers and workers are significantly different from one another and from nymphs and alates. Nineteen peaks are statistically significant between workers and soldiers. Nymphs and alates were not statistically different. We detected statistically significant quantitative differences between colonies in 18 peaks for workers and 12 peaks for soldiers. Each of the colonies ofC. formosanus can be separated from the others by the proportions of their hydrocarbon components. We detected statistically significant differences between months of the year for 12 peaks for workers and four peaks for soldiers; two peaks each for workers and soldiers showed distinct, seasonal trends. This seasonal shift in proportions of hydrocarbons correlates with the production of alates.     

Identification of trail pheromone precursors from Subterranean termite,Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae)

Whole-body extracts of the termite,Coptotermes formosanus Shiraki served for examining the presence of trail pheromone precursor(s). Three trail pheromone precursor candidates, identified as dodecatrienyl stearate, dodecatrienyl oleate, and dodecatrienyl linoleate, were isolated using various chromatographic methods in conjunction with bioassay and by capillary GC-MS analyses.     

Cuticular hydrocarbons of eight species of north american cone beetles,Conophthorus hopkins

A study to determine the degree of similarity and/or diversity among eight of the 15 described species ofConophthorus is reported. Cuticular hydrocarbons were evaluated forC. conicolens, C. ponderosae, C. cembroides, C. edulis, C. radiatae, C. coniperda, C. resinosae, andC. banksianae. Seventy-eight individual and isomeric mixtures of hydrocarbons were identified by gas chromatography-mass spectrometry, includingn-alkanes, alkenes, alkadienes, 2- or 4-methylalkanes, 3-methylalkanes, and single-component and isomeric mixtures of internally branched mono-, di-, and trimethylalkanes. Differences in alkenes and mono-, di-, and trimethylalkanes can be used easily to separate the eight species.Conophthorus conicolens andC. ponderosae contain the most complex blends. Hydrocarbon patterns in three geographically separated populations ofC. ponderosae, each from a different host, are qualitatively identical with the exception of a homologous series of 3,7-dimethylalkanes from adults collected fromPinus lambertiana cones. The latter could comprise a sibling species. Hydrocarbon mixtures of two eastern species,C. resinosae andC. banksianae, are qualitatively identical, supporting the suspicion thatC. banksianae may not be a valid species. Closely relatedC. cembroides andC. edulis have similar combinations of hydrocarbons except for a unique and abundant alkene (C₂₇₁) inC. edulis and two dimethyhexacosanes inC. cembriodes.Coleoptera: Scolytidae.     

(Z,E,E)-dodecatrien-1-ol: A minor component of trail pheromone of termite,Coptotermes formosanus Shiraki

In the course of the elucidation of the primary structure of an isolated trail pheromone fromC. formosanus, a minor component that had the same molecular weight as the major trail pheromone, (Z,Z,E)-3,6,8-dodecatrien-1-ol [(Z,Z,E)-DTE-OH], was detected in the mass chromatogram ofm/z 180 of capillary GC-MS. The mass spectrum of the minor component showed a prominent pattern of dodecatrien-1-ol. Chemical analysis demonstrated that the complete structure was (Z,E,E)-DTE-OH. Furthermore, capillary GC-MS-HR-SIM analysis indicated that the component existed only in the workers ofCoptotermes formosanus Shiraki and was not present in workers ofReticulitermes speratus (Kolbe). This minor component may be a species-specific factor ofC. formosanus, although this was not suggested by a two-choice bioassay.     

Trilinolein Identified as A Sex-Specific Component of Tergal Glands in Alates of Coptotermes formosanus

Hexane extracts of the tergal glands from female alates of the Formosan subterranean termite Coptotermes formosanus were analyzed by high performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry with collision-induced dissociation. Double bond configuration was determined by chemical modifications with gas chromatography-mass spectrometry. A single component, identified as the triacylglycerol, trilinolein, was unique to the female tergal glands. This compound was not found in other areas of the female alate abdomen or in the corresponding area of male alates. Neither gland extract nor trilinolein caused a behavioral response from male alates. However, significant differences were found between males and females for responses from neurons within sensilla of the maxillary palps.     

Allelochemical resistance of bald cypress,Taxodium distichum,heartwood to the subterranean termite,Coptotermes formosanus

The heartwood of bald cypress,Taxodium distichum (L.) Rich., resisted feeding attack by the Formosan subterranean termite,Coptotermes formosanus Shiraki. Hexane-extracted heartwood, however, was consumed at > 12 times the amount of sound heartwood eaten. A bioassay usingT. distichum sapwood as a feeding substrate was employed to assess the antitermitic activity of successive hexane, acetone, and methanol extracts of heartwood shavings and isolates derived from the active hexane extract. Two fractions, eluted from the crude hexane extract by liquid chromatography, significantly reduced termite feeding compared to the parent extract, while a third fraction was less active than the original hexane extract. Each fraction contained one major component. All three components were structurally related diterpenes. The two most active heartwood constituents were identified by GC-MS and NMR as ferruginol and manool, while the third and least active, but most prevalent, compound in heartwood was identified as nezukol. Results of bioassays suggest that these allelochemicals act principally as feeding deterrents with accompanying termite mortality due to starvation.     

Determination of feeding preference of Formosan subterranean termite (Coptotermes formosanus Shiraki) for some amino acid additives

A choice feeding test using 21 amino acids was conducted to determine the feeding preference of Formosan subterranean termite,Coptotermes formosanus Shiraki, in the laboratory. Significantly more filter paper treated withd-aspartic acid orl-glutamic acid was consumed by Formosan subterranean termites than was control filter paper treated with water. In two-choice feeding tests, termites consumed significantly more filter paper treated withd-aspartic acid orl-aspartic acid than paper treated with water. Addingl-proline,l-lysine, orl-isoleucine to filter paper significantly increased consumption compared with control filter paper in no-choice tests. The use of amino acid additives in termite baits is briefly discussed.     

Cuticular Hydrocarbons and Soldier Defense Secretions of <Emphasis Type="Italic">Reticulitermes</Emphasis> in Southern California: A Critical Analysis of the Taxonomy of the Genus in North America

Cuticular hydrocarbons (CHC) and soldier defense secretions (SDS) were characterized for collections of Reticulitermes from six counties (Los Angeles, Orange, Riverside, San Bernardino, San Diego, and Santa Barbara) in southern California. Collection sites included the type locality for R. hesperus, Lake Arrowhead (formerly known as Little Bear Lake) in the San Bernardino Mountains. In southern California, there are two CHC phenotypes, SC-A and SC-B, which are easily distinguished by the presence or absence of 5-methyl pentacosane, 5-methyl heptacosane, 5,17-dimethyl pentacosane, and 5,17-dimethyl heptacosane. These phenotypes are similar, but not identical, to previously designated phenotypes of Reticulitermes from northern California. The SDS of termites collected from southern California were characterized; (—)-germacrene A was abundant in all but the four samples from Lake Arrowhead. Soldiers of phenotype SC-A produced >79% germacrene A. The four samples from Lake Arrowhead produced no germacrene A, but contained >78% γ-cadinene. The SDS from the Lake Arrowhead samples were more similar to those of CA-A/CA-A′ from northern California than to any of the CHC phenotypes from southern California. Soldiers of CHC phenotype SC-B produced germacrene A, with the proportion varying from 16.2 to 98.7%. The SDS of SC-B were more similar to those of SC-A than to any of the phenotypes from northern California. The CHC phenotype SC-A found in southern California likely represents R. hesperus and SC-B appears to be a new, as yet undescribed species. We discuss the state of current taxonomic research on Reticulitermes.

Lignoceric Acid and Hexacosanoic Acid: Major Components of Soldier Frontal Gland Secretions of the Formosan Subterranean Termite (Coptotermes formosanus)

Qualitative and quantitative analyses were performed on trimethylsilyl derivatives of soldier frontal gland secretions of the Formosan subterranean termite, Coptotermes formosanus Shiraki, by using gas chromatography–mass spectrometry. Ranges of 17.23–33.78 mg/g (acid/wet weight of secretion) of lignoceric acid and 5.39–7.67 mg/g of hexacosanoic acid were found in the soldier frontal gland secretion of three different colonies. This is the first report that organic acids occur in the soldier secretions of this species.     

Cuticular hydrocarbons of the eastern subterranean termite,Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae)

Major cuticular hydrocarbon components in several castes ofReticulitermes flavipes (Kollar) have been identified and quantitated. Types of hydrocarbons present includen-alkanes, 2-methylalkanes, 3-methylalkanes, 5-methylalkanes, an alkene, and an alkadiene, with a range in carbon numbers from C₂₁ to C₂₆, This is the first report on insect cuticular hydrocarbons in which both 2- and 3-methylalkanes are present, as well as the first report of an insect with a conjugated alkadiene.     

How Reliable is the Analysis of Complex Cuticular Hydrocarbon Profiles by Multivariate Statistical Methods?

Numerous recent studies have correlated cuticular hydrocarbon profiles with a wide range of behaviors, particularly in social insects. These findings are wholly or partly based on multivariate statistical methods such as discriminate analysis (DA) or principal component analysis (PCA). However, these methods often provide limited insight into the biological processes that generate the small differences usually detected. This may be a consequence of variability in the system due to inadequate sample sizes and the assumption that all compounds are independent. A fundamental problem is that these methods combine rather than separate the effects of signal components. By using cuticular hydrocarbon data from previous social insect studies, we showed that: (1) in 13 species of Formica ants and seven species of Vespa hornets, at least one group of hydrocarbons in each species was highly (r ² > 0.8) correlated, indicating that all compounds are not independent; (2) DA was better at group separation that PCA; (3) the relationships between colonies (chemical distance) were unstable and sensitive to variability in the system; and (4) minor compounds had a disproportionately large effect on the analysis. All these factors, along with sample size, need to be considered in the future analysis of complex chemical profiles.     

Cuticular hydrocarbons of adults of the cowpea weevil,Callosobruchus maculatus

The composition of the cuticular hydrocarbons of the cowpea weevil,Callosobruchus maculatus (F.), was determined by using combined gas chromatography-mass spectrometry. The hydrocarbons constituted 88% of the cuticular lipids and were composed of four homologous series of alkanes. Mono- and dimethyl branched-chain alkanes made up 83% of the hydrocarbon fraction.Coleoptera: Bruchidae.Mention of a proprietary product or company name in this paper does not constitute a recommendation of this product by the U.S. Department of Agriculture.     

Cuticular hydrocarbons of the paper wasp,Polistes fuscatus: A search for recognition pheromones

The cuticular chemicals of 124 individual wasps (foundresses and workers) from 23 colonies ofPolistes fuscatus were analyzed. The compounds identified, all of which were hydrocarbons, were similar to those of other vespid wasps in that the bulk of the hydrocarbons were 23–33 carbons in chain length. However, the hydrocarbon profile ofP. fuscatus differed from those of its congeners in its proportions of straight-chain alkanes, methylalkanes, and alkenes. Three of the 20 identified hydrocarbons, 13- and 15-MeC₃₁, 11,15- and 13,17-diMeC₃₁, and 13-, 15-, and 17-MeC₃₃, had properties postulated for recognition pheromones: colony specificity, efficacy in assigning wasps to the appropriate colony, heritability, lack of differences between foundresses and workers, and distinctive stereochemistry.     

Task-Related Differences in the Cuticular Hydrocarbon Composition of Harvester Ants, Pogonomyrmex barbatus

Colonies of the harvester ant, Pogonomyrmex barbatus, perform a variety of tasks. The behavior of an individual worker appears to depend on its recent history of brief contacts with ants of the same and other task groups. The purpose of this study was to determine whether task groups differ in cuticular hydrocarbon composition. We compared the cuticular hydrocarbon composition of ants collected under natural conditions as they performed one of three tasks: patrolling (locating food sources), foraging, or nest maintenance. Task groups differed significantly in the relative proportions of classes of hydrocarbon compounds, as well as in individual compounds. Relative to nest maintenance workers, foragers and patrollers had a higher proportion of straight-chain alkanes relative to monomethylalkanes, dimethylalkanes, and alkenes. There was no significant difference in the chain length of n-alkanes among the task groups. Foragers did not differ in hydrocarbon composition from patrollers. Colonies differed significantly from one another in hydrocarbon composition, but task groups differed in consistent ways from colony to colony, suggesting that the mechanism responsible for task-related hydrocarbon composition was the same in all colonies. P. barbatus workers switch tasks during their lifetimes, suggesting that cuticular hydrocarbon composition changes during adulthood as well. Nest maintenance workers are probably younger than foragers and patrollers and perform very little of their work outside of the nest. Task-related hydrocarbon differences detected here may be associated with worker age, and/or the abiotic characteristics (temperature, humidity, and ultraviolet light) of the interior and exterior work environments.     

Cuticular Hydrocarbons Provide Reliable Cues of Fertility in the Ant Gnamptogenys striatula

In ca. 150 species of queenless ants, a specialized queen caste is rare or absent, and mated workers take over the role of the queen in some or all of the colonies. Previously, it has been shown that reproduction in queenless ants is regulated by a combination of dominance behavior and chemical fertility signaling. It is unknown, however, whether chemical signals alone can sufficiently regulate reproduction. To investigate this possibility, we studied reproductive regulation in the facultatively queenless ant Gnamptogenys striatula, a species where dominance behavior is rare or absent. Active egg layers and infertile workers showed qualitative and quantitative differences in their cuticular hydrocarbon profile. Five long-chain methyl alkanes, 3,13- and 3,15-dimethyl pentriacontane, 3,13- and 3,15-dimethyl heptentriacontane, and 3,11,15-trimethyl heptentriacontane occurred only on the cuticles of virgin and mated egg layers. Pronounced quantitative differences were found in a further 27 alkenes; alkanes; and mono-, di-, and trimethyl alkanes. Workers that had recently stopped laying eggs had profiles similar to infertile workers, and mating status did not affect this chemical pattern. We conclude that the cuticular hydrocarbon profiles of G. striatula workers provide reliable information about their current fertility. In the interest of colony productivity, this allows reproduction to be regulated without the use of aggression.     

Cuticular Hydrocarbons of Polistes dominulus as a Biogeographic Tool: A Study of Populations from the Tuscan Archipelago and Surrounding Areas

In social insects, the types and proportions of epicuticular lipids may exhibit significant diversity as a result of factors such as age, sex, caste, rank, nest, and relatedness. It is known that these variations can be used by social insects to acquire information regarding conspecific individuals. Recent findings have shown that different populations of Polistes dominulus (Christ.) have distinctly different chemical cuticular profiles, and that wasps are able to recognize individuals of their own population. In this study, we showed that cuticular hydrocarbon patterns of Polistes dominulus are consistent with similarities among northern Tyrrhenian islands, as reported in previous biogeographic studies. Indeed, our findings indicate that cuticular hydrocarbon mixtures of P. dominulus from Capraia and Corsica are grouped together by cluster analysis, while those from Elba and Giglio cluster with cuticular profiles of the mainland wasps (Venturina).     

Cuticular Hydrocarbons as Chemotaxonomic Characters of Pine Engraver Beetles (Ips spp.) in the grandicollis Subgeneric Group

Cuticular hydrocarbons were extracted, identified, and evaluated as chemotaxonomic characters from all species of adult Ips pine engraver beetles in the grandicollis subgeneric group. The grandicollis group consists of Ips grandicollis (Eichhoff), I. cribricollis (Eichhoff), I. lecontei Swaine, I. montanus (Eichhoff), I. paraconfusus Lanier, I. confusus (LeConte), and I. hoppingi Lanier. In order to provide outgroups for a phylogenetic analysis, cuticular hydrocarbons were also analyzed from Orthotomicus caelatus (Eichhoff), I. latidens (LeConte) (latidens subgeneric group), and I. pini (Say) (pini subgeneric group). Two hundred forty-eight hydrocarbon components were identified by gas chromatography–mass spectrometry. The members of the grandicollis group provided 206 of these compounds. The components represented eight classes: n-alkanes, alkenes, alkadienes, terminally branched methylalkanes, internally branched methylalkanes, dimethylalkanes, trimethylalkanes, and tetramethylalkanes. Different populations of O. caelatus, I. grandicollis, I. lecontei, I. montanus, I. paraconfusus, I. confusus, and I. hoppingi provided no evidence for interpopulational variation in cuticular hydrocarbons. Single populations only were analyzed for I. latidens, I. pini, and I. cribricollis. Sexual dimorphism in cuticular hydrocarbons occurred only in I. lecontei where females produced eight unique components with a pentatriacontane parent chain. Several phylogenetic analyses based on hydrocarbon phenotypes agreed in general with the established morphologically based system of relatedness and with published phylogenies reconstructed from protein and nucleic acid characters. Nearly all hydrocarbon analyses suggested a close relationship between I. grandicollis and I. cribricollis; between I. lecontei and I. montanus; and among the sibling species I. paraconfusus, I. confusus, and I. hoppingi. The presence or absence of specific n-alkanes (n-docosane, n-triacontane); certain dimethylalkanes (terminally branched with octacosane and triacontane parent chains and internally branched with heptacosane, hentriacontane, and docotriacontane parent chains); and 3,7,11-; 3,7,15-trimethylheptacosane permit facile discrimination of I. paraconfusus, I. confusus, and I. hoppingi. These three sibling species are difficult to resolve by external morphology. These data support the species status of I. hoppingi rather than it being considered a host race of the I. confusus complex. They also support the species status of I. cribricollis rather than it being considered part of I. grandicollis. In contrast to other published phylogenies reconstructed from molecular data, phylogenies reconstructed from cuticular hydrocarbons repeatedly place I. lecontei as an integral part of the grandicollis subgeneric group. Thus, cuticular hydrocarbon and pheromone alcohol composition of I. lecontei support its inclusion in the grandicollis subgeneric group.     

Trail-following pheromones of the rhinotermitidae: Approaches to their authentication and specificity

A method for determining the authenticity of subterranean termite trail pheromones is suggested and utilized to verify the presence of trail pheromones inReticulitermes virginicus, R. flavipes, andR. tibialis. In addition, a possible trail pheromone has been demonstrated forCoptotermes formosanus. A choice bioassay method shows that the above trail pheromones are species specific.     

Chemometric Classification of Comb and Cuticular Waxes of the Honeybee Apis Mellifera Carnica

Waxes are important as building material and for the chemical communication of the honeybee Apis mellifera carnica. In this study chemometric tools were established for classifying the different waxes inside the hive. By using gas chromatography in combination with mass spectrometry, components of different types of waxes were analyzed. By considering different substance classes of waxes, discriminant function analyses revealed distinct subtypes of comb waxes and of cuticular waxes. It is shown that the aging of comb wax is in part a spontaneous physicochemical process due to differential volatilities of compound classes with different chain length ranges. On the other hand it is directly influenced by the bees by adding lipolytic enzymes to the comb wax. The data suggest that the varying cuticular wax and comb wax compositions could serve as cues for bees to recognize castes, sexes, or comb age.     

Changes in the Cuticular Hydrocarbon Profile of the Slave-Maker Ant Queen, Polyergus breviceps Emery, After Killing a Formica Host Queen (Hymenoptera: Formicidae)

Queens of the slave-maker ant, Polyergus breviceps, take over nests of their Formica host species by fatally attacking the resident queen. As workers only begin grooming the P. breviceps queen once she has ceased her attack, we investigated whether a change in parasite queen chemistry may account for the change in worker behavior. Cuticular hydrocarbon profiles of newly mated P. breviceps queens and of queens of their two Formica host species were found to be species-specific. Profiles of newly mated P. breviceps queens that had attacked a Formica queen, however, were virtually identical to the queen profile of the species killed. Mass spectral analysis revealed that the hydrocarbons on the cuticles of newly mated P. breviceps changed from primarily normal alkanes to methyl and di-methyl branched alkanes after attacks. The results suggest that cuticular compounds from the host queen were transferred to the parasite queen during their aggressive interaction.