Pattern of recovery of species-specific cuticular hydrocarbon mixtures by Reticulitermes santonensis and Reticulitermes lucifugus grassei after being removed from a mixed group. Is the acquisition of allospecific hydrocarbons reversible?

Each of the termite species Reticulitermes santonensis and Reticulitermes lucifugus grassei has its own particular cuticular chemical profile. When members of the two species are placed together to form artificially mixed species groups, their chemical profiles undergo changes: Each species acquires all the hydrocarbons which initially characterized the other species. When the members of a mixed group which had been kept together for 24 h were split into two homospecific groups, the cuticular profiles of the members of both groups immediately showed a sharp drop in both the homospecific and allospecific components. In R. santonensis, the homospecific hydrocarbons subsequently increased in quantity, reaching values which were higher on the 33rd day after the separation than those initially recorded in this species; whereas in R. lucifugus grassei, the homospecific hydrocarbon proportions were still lower on the 33rd day than the initial values. In both species, the allospecific hydrocarbon levels began to increase sharply on the 5th day after separation, and the homospecific products still showed no tendency to return to the initial proportions 33 days after separation. In the light of these results, some hypotheses are put forward as to what mechanisms might posibly regulate the hydrocarbon profiles of these two species. Arch. Insect Biochem. Physiol. 35:237-259, 1997.© 1997 Wiley-Liss, Inc.     

Phylogenetic analyses of two mitochondrial genes and one nuclear intron region illuminate European subterranean termite (Isoptera: Rhinotermitidae) gene flow, taxonomy, and introduction dynamics

Phylogenetic analyses of multiple DNA sequences were conducted to elucidate gene flow, evolutionary patterns, taxonomy, and the dynamics of two accidental introductions: Reticulitermes lucifugus grassei into Devon, United Kingdom and R. flavipes into Europe. Two mitochondrial DNA genes totaling 1495 bp and a 380-bp ribosomal intergenic transcribed spacer were sequenced. Neighbor-joining and parsimony analyses revealed that multiple female lineages of R. lucifugus grassei were introduced into Devon possibly from southwestern France, where the species was indigenous. The taxonomic status of the European R. santonensis as a species separate from the North American R. flavipes has been questioned since it was described in 1924. Phylogenetic analyses revealed a close genetic relationship between R. flavipes from the United States and R. santonensis from France. These analyses, coupled with morphological and chemotaxonomic data, provide strong support for R. santonensis and R. flavipes being the same species. They also suggested that R. santonensis infestations likely resulted from R. flavipes being introduced into Europe.     

Phylogeography of two European Reticulitermes (Isoptera) species: the Iberian refugium

Chemical, i.e. cuticular hydrocarbons, and molecular data were used to probe the phylogeography of Reticulitermes termites collected from various parts of France, Spain and Portugal. Phylogenetic relationships were inferred from sequences of the internal transcribed spacer (ITS2) of nuclear ribosomal RNA genes as well as from two partial mitochondrial DNA segments, the cytochrome oxidase II gene and a sequence combining the tRNA-Leu gene and fragments of the NADH dehydrogenase I and ribosomal 16S genes. Two species, namely, R. grassei and R. banyulensis, were identified based on an analysis of cuticular hydrocarbons and the identification was confirmed by ITS2 haplotyping. However, phylogeny based on the analysis of mitochondrial DNA was not completely in agreement with the conclusions drawn from the chemical and nuclear data. An analysis of 56 R. grassei colonies revealed intraspecific differentiation into two major lineages with distinct geographical ranges. Whereas analysis of cuticular hydrocarbons showed that R. banyulensis was chemically distinct from R. grassei, analysis of mitochondrial DNA showed its close kinship with the R. grassei lineage occurring in southern Spain. This kinship could be explained by their evolution from a common polymorphic ancestor species in this ice age refugium.     

Molecular relationships among European samples of Reticulitermes (Isoptera, Rhinotermitidae)

Taxonomy and phyletic relationships of the European termites of the holarctic genus Reticulitermes are highly debated and poorly known. Sequencing analyses of the complete cytochrome oxidase subunit II and of a fragment of 16S rDNA mitochondrial genes were performed on 21 Italian and French populations. Distance, parsimony, and maximum-likelihood evaluations on single and combined data sets confirm the presence in central peninsular Italy of R. lucifugus lucifugus, but demonstrate that a coastal Tuscan population and the Sardinian ones pertain to the Corsican subspecies R. lucifugus corsicus, whose distribution appears therefore transtyrrenian. Northeastern Italian samples are highly differentiated from presently analyzed R. lucifugus and constitute a new entity, possibly more related to the Japanese R. speratus. Finally, the French R. santonensis and the North American R. flavipes appear strongly related, so that the supposed synonymy between the two taxa must be accepted.     

Comparison of the spatial distribution and reproductive cycle of Reticulitermes santonensis Feytaud and Reticulitermes lucifugus grassei Clément (Isoptera, Rhinotermitidae) suggests that they represent introduced and native species, respectively

Summary: The termites Reticulitermes santonensis (Rs) and R. lucifugus (Rl) were first regarded as different species (Feytaud, 1924) and then as the same species (Buchli, 1958). In later studies, Clément (1978, 1986) and Bagnères et al. (1990) used morphological and chemical (cuticular hydrocarbons) criteria to show that they actually are two distinct species. Feytaud (1924) and Bagnères et al. (1990) suggested that Rs could have originated from a North American population of R. flavipes that was introduced into Europe. The present study examined the distribution and reproductive cycle of Rs and an Rl sup-species, Rl grassei (Rlg), which are partly sympatric in southwest France. In the wild, Rs has an aggregated distribution related to human surroundings (past or present), i.e. populations are either dense or entirely absent. Conversely, Rlg is distributed randomly (from dense to few) throughout its range. For example, Rs is not found in towns within the Landes Forest, an area in southwest France that remained economically depressed and sparsely settled until after 1850 (Sargos, 1997), while Rlg is quite common there. Their respective reproductive strategies show that Rs populations spread through budding (swarms are scarce and many brachypterous nymphs and neotenics are found), whereas Rlg populations reproduce by both swarming and budding. It is likely that Rs is an introduced species and Rlg a native one that has reached its northern limits in France. A closer examination of North American Reticulitermes species, particularly R. flavipes, might reveal a relationship with the European species.     

Distribution and genetic variation of Reticulitermes (Isoptera: Rhinotermitidae) in Portugal

The genus Reticulitermes has a Holartic distribution and is widely represented in southern Europe. It is established in Portugal, across the full range of ambient environmental conditions, using different types of wood resources, as available, and nesting ubiquitously in many types of soil. Sequencing of part of the mtDNA COII gene revealed that all the specimens sampled on a north-south transect of the country were Reticulitermes grassei (considered synonymous with R. lucifugus grassei). Nevertheless, an association between the genetic and the geographical distances of sampled specimens was shown, leading to the conclusion that discrete termite populations have differentiated in situ. The geographic component of the genetic polymorphism should be the subject for further research.     

Ontogenic potentialities of the worker caste in two sympatric subterranean termites in France

In termites, the capacity of workers to differentiate into neotenic reproductives is an important characteristic that deserves particular attention. To gain insight into the differentiation pathway, the potentialities of workers and the endocrinal changes during the formation of neotenics were compared in two sympatric termites, Reticulitermes flavipes and Reticulitermes grassei. After 1 year of development, 100% of R. flavipes worker groups produced neotenics against only 63% of R. grassei groups. The average production of female neotenics was significantly higher in R. flavipes worker groups compared with R. grassei groups and R. flavipes produced a greater proportion of female neotenics. Moreover, R. flavipes produced more offspring, not only because there were more females, but also because R. flavipes females were more productive. Moreover, the offspring produced by R. flavipes grew faster than the offspring of R. grassei. Both ecdysteroid and juvenile hormone (JH) titers varied significantly during the development of neotenics. The two species showed similar ecdysteroid titer variation patterns. However, the JH titer variation patterns strongly differed: in R. grassei, the concentration of JH increased in maturing neotenics then dropped in mature neotenics, whereas in R. flavipes, the level of JH was significantly higher than in R. grassei and remained constantly high in mature neotenics. Overall, these results suggest that these two species differ strongly in many life-history traits as well as in the physiological control of their caste differentiation system. Possible origins and mechanisms of such interspecific variations are discussed, as well as their evolutionary and ecological consequences.     

Characterization of termite lipophorin and its involvement in hydrocarbon transport

The transport of lipids constitutes a vital function in insects and requires the plasma lipoprotein lipophorin. In all insects examined to date, cuticular hydrocarbons are also transported through the hemolymph by lipophorin, and in social insects they play important roles not only in water proofing the cuticle but also in nestmate recognition. High-density lipophorin (HDLp), isolated from Reticulitermes flavipes plasma by KBr gradient ultracentrifugation, contains 66.2% protein and 33.8% lipids; hydrocarbons constitute its major neutral lipid (20.4% of total lipids). Anti-lipophorin serum was generated in rabbit and its specific association with lipophorin, and not with any other plasma proteins, was verified with Western blotting. Immunoprecipitation also confirmed that this antibody specifically recognizes lipophorin, because all hemolymph hydrocarbons of the termites R. flavipes and R. lucifugus and the cockroach Supella longipalpa, which associate only with lipophorin, were recovered in the immunoprecipitated protein. Cross-reactivity of the antiserum with lipophorin from related species was investigated by double immunodiffusion with 10 termite species in the genera Reticulitermes, Coptotermes, Zootermopsis, and Kalotermes, and with five cockroach species. Involvement of lipophorin in hydrocarbon transport was shown by injecting HDLp antiserum into Zootermopsis nevadensis and then monitoring the de novo biosynthesis of hydrocarbons and their transport to the cuticular surface; the antiserum significantly disrupted hydrocarbon transport. ELISA revealed a gradual increase in the lipophorin titer in successively larger R. flavipes workers, and differences among castes in lipophorin titers were highest between nymphs and first instar larvae.     

Phylogenetic relationships of nearctic Reticulitermes species (Isoptera: Rhinotermitidae) with particular reference to Reticulitermes arenincola Goellner

DNA sequence comparisons of the mitochondrial COII, 16S, and 12S rRNA genes were used to infer phylogenetic relationships among the six known US Reticulitermes species (Reticulitermes flavipes, Reticulitermes arenincola, Reticulitermes tibialis, Reticulitermes hageni, Reticulitermes virginicus, and Reticulitermes hesperus) and the closely related European species Reticulitermes santonensis. The interspecific pairwise sequence divergence, based on uncorrected "p" distance, varied up to 10% across the COII, 4.9% across the 16S, and 3% across the 12S fragments. Phylogenetic trees were constructed using maximum parsimony, likelihood, and distance methods. The combined results suggest several phylogenetic relationships including: (i) R. flavipes, R. arenincola, and European R. santonensis are possibly conspecific; (ii) R. virginicus and R. hageni are closely related species; and (iii) R. tibialis and R. hesperus are closely related species. Interestingly, while there is apparent synonymity between R. flavipes and R. arenincola by DNA sequence, there are clear morphological differences in the soldier caste. This finding suggests a combination of molecular and morphological approaches are necessary for accurate species identification. These data lend resolution to the complex problem of Reticulitermes systematics, and will assist future efforts directed toward characterizing species distribution and ecology.     

Alarm Reaction of European Reticulitermes Termites to Soldier Head Capsule Volatiles (Isoptera, Rhinotermitidae)

The alarm behavior of worker and soldier castes of four European subterranean termite species of the genus Reticulitermes (R. santonensis, R. lucifugus, R. grassei, R. banyulensis) is described. In petri dish bioassays we presented filter papers with squashed soldier heads as an odor source to trigger a alarm reaction. Untreated filter papers were used as controls. The behavioral patterns were alike for all four species. In the control situation (no alarm), only occasional workers and few soldiers were seen walking in the petri dish. When the odor source was presented, a general state of alertness was observed, showing as higher activity and presence, zigzag running, antennation of nestmates, jittering and jerking, mandible snapping, head-banging, and attraction to the odor source. Workers were attracted within a few seconds (6.9–13.4 s); soldiers followed much later (18.5–64.6 s). Both workers and soldiers inspected the odor source only briefly (workers for 8.7–12.8 s, soldiers for 10.0–19.5 s), with the exception of R. santonensis, where soldiers stayed an average of 209.8 s. The number of individuals, of 50 workers and 3 soldiers, near the odor source averaged 6.0–9.2 workers and 1.2–2.3 soldiers. The presence of workers is crucial: without workers, a significant alarm reaction in soldiers could not be induced. The roles of soldier and worker castes of Reticulitermes species within alarm communication and the origin and nature of possible alarm signals are discussed.     

Origin of a new Reticulitermes termite (Isoptera, Rhinotermitidae) inferred from mitochondrial and nuclear DNA data

The Holoarctic termite genus Reticulitermes is widely distributed in Europe. A new Reticulitermes species, R. sp. nov, was recently found in France and Italy. Its phylogenetic position was investigated using a 743-bp fragment of mitochondrial 16S rRNA-ND1 genes and 382-bp of the nuclear ITS2 region. Phylogenies for these sequences were estimated by neighbor-joining, maximum-parsimony and maximum-likelihood analysis. The results strongly supported a relationship between R. sp. nov. and the termite species from the eastern Mediterranean area including Reticulitermes balkanensis from the Balkans, Reticulitermes lucifugus from Turkey and Reticulitermes clypeatus from Israel. The hypothesis of a relationship between R. sp. nov. and the Japanese Reticulitermes speratus was rejected by parametric bootstrap. The current distribution of R. sp. nov. could be linked to postglacial colonization routes between Balkan refuge and northern regions.     

Nestmate discrimination and cuticular profiles of a temporary parasitic ant Lasius sp. and its host L. fuliginosus (Hymenoptera, Formicidae)

Workers of the temporary parasitic ant Lasius sp. were perfectly compatible with their hosts Lasius fuliginosus. Aggression was never seen between allospecific nestmates in either field or laboratory. In the laboratory, trophallaxis and allogrooming between allospecific nestmates were statistically more frequent than that between conspecifics. These ants were highly aggressive toward individuals of either species from another mixed colony located 1 km away, indicating that they discriminated nestmates and nonnestmates regardless of the species to which they belonged. No aggressive actions, however, were observed between two neighboring mixed nests located 5 m apart. We have not considered, however, the relationship of the two neighboring nests: the two nests may have been two subunits of the same family or sister colonies, or may have been different colonies but discriminated from distant aliens. Gas chromatography analysis on the total cuticular extracts showed that Lasius sp. and L. fuliginosus possess distinct cuticular profiles, even though they were collected from the same mixed colony. Received: December 13, 1999 / Accepted: April 25, 2000     

Changes of species specific cuticular hydrocarbon profiles in the cockroaches Nauphoeta cinerea and Leucophaea maderae reared in heterospecific groups

The cuticular hydrocarbon profiles of the cockroaches Nauphoeta cinerea and Leucophaea maderae are species-specific when maintained in homospecific rearings. When individuals were reared in mixed species colonies, they initially remained in homospecific groups under different shelters. However, after 14 days they formed one heterospecific group with cuticular profiles showing characteristics of both species. When individuals were returned in monospecific rearings, their cuticular hydrocarbon profiles returned to species-specific ones within 3 weeks.     

Coevolution of symbiotic spirochete diversity in lower termites.

The phylogenetic relationships of symbiotic spirochetes from five dry-wood feeding lower termites (Cryptotermes cavifrons, Heterotermes tenuis, Kalotermes flavicollis, Neotermes mona, and Reticulitermes grassei) was compared to those described in previous reports. The 16S rDNA bacterial genes were PCR-amplified from DNA isolated from intestinal samples using a spirochete-selective primer, and the 16S amplicons were cloned into Escherichia coli. Sequences of the cloned inserts were then used to determine closest relatives by comparison with published sequences. Clones sharing more than 97% sequence identity were grouped into the same phylotype. Forty-three new phylotypes were identified. These termite whole-gut-spirochetes fell into two previous defined clusters, designated as Treponema Clusters I and II, and one new Cluster III. Thirty-seven phylotypes were grouped in Cluster I. Cluster II comprised three phylotypes, two from Reticulitermes grassei (LJ029 and LJ012) and one from Heterotermes tenuis (LQ016). Three phylotypes, LK057, LK050 and LK028, were affiliated to Cluster III. Members of Cluster I showed the following characteristics: (i) spirochete phylotypes from a particular species of termite were more closely related to each other than to phylotypes of other termite species; (ii) spirochetes obtained from different genera of the same family, such as Cryptotermes sp., Kalotermes sp., and Neotermes sp., all from the family Kalotermitidae, were also related to each other. It was therefore concluded that spirochetes are specific symbionts that have coevolved with their respective species of termites, are stably harbored, and are closely related to members of the same termite family.     

Response of Reticulitermes spp. (Isoptera: Rhinotermitidae) in northern California to baiting with hexaflumuron with sentricon termite colony elimination system

Colonies of Reticulitermes spp. were baited with prototype and commercial Sentricon stations (Dow AgroSciences LLC, Indianapolis, IN) to test the efficacy of hexaflumuron in different concentrations and bait matrices and to document reinvasion of the foraging territories vacated by eliminated colonies. Seven colonies of Reticuliternes spp. from two sites were characterized with cuticular hydrocarbon analyses and mark-release-recapture and agonistic behavioral studies. Three colonies were observed as controls and four colonies were baited. When a connection between the bait station and the monitoring station could not be confirmed by mark-release-recapture studies, the results of the baiting were equivocal. The monitoring stations of a colony at our wildland site were devoid of termites 406 d after baiting with one Sentricon station, but became reoccupied with the same species of termites approximately 6 mo after baiting. A colony at the residential site was baited with 0.5% hexaflumuron in the Recurit II bait matrix; 60 d later termites were absent from all monitoring stations. These monitoring stations remained unoccupied for > or = 18 mo. Foraging Reticulitermes spp. appeared in three of the seven monitoring stations 18, 24, and 36 mo after baiting, respectively. Using cuticular hydrocarbon analyses and agonistic behavior studies, we determined that the Reticulitermes spp. occupying these monitoring stations were from three different colonies; none were members of the original colony destroyed by baiting. Another colony at the residential site was baited using a noncommercial, experimental bait; 52 d later termites were absent from all monitoring stations. The monitoring stations remained unoccupied for > or = 9 mo. A different Reticulitermes sp. colony invaded one monitoring station 9 mo after baiting.     

Cuticular Hydrocarbons of Reticulitermes (Isoptera: Rhinotermitidae) from Northern California Indicate Undescribed Species

Cuticular hydrocarbon mixtures of Reticulitermes samples from disparate locations in northern California were characterized and correlated with species determinations. Literature records indicate that there are two extant species of Reticulitermes in northern California: Reticulitermes hesperus Banks and R. tibialis Banks. We identified five distinct hydrocarbon phenotypes in Reticulitermes from northern California. Termites were diagnosed morphologically as R. tibialis for all five cuticular hydrocarbon phenotypes based on available keys to soldiers. Commonly accepted distributions of Reticulitermes in northern California indicate that our primary collection sites should only produce R. hesperus. On the basis of cuticular hydrocarbons alone we suggest that there are two or more undescribed taxa of Reticulitermes in northern California, or that described species of Reticulitermes occur in northern California, but have never been noticed before. Therefore, we conclude that the taxonomy of Reticulitermes in North America is in need of revision.     

白蚁表皮碳氢化合物组分鉴定及分类学意义

应用GC-MS分析表明,不同种类白蚁表皮碳氢化合物组成和含量均有差异。运用UPGMA聚合R分析的最小距离系数值绘制的系统树表明：圆唇凸额类白蚁——黄胸散白蚁Reticulitermes flaviceps、双色散白蚁R. dichrous和R. sp.1之间;圆唇平额类白蚁——圆唇散白蚁R. labralis、小头散白蚁R. microcephalus、R. sp.2和R. sp.3之间; 尖唇类白蚁——海南异白蚁Heterotermes hainanensis、拧黄异白蚁H. citrinus、细颚异白蚁H.leptomandibularis、尖唇异白蚁H. aculabialis和H. sp. 4之间的亲缘关系较近。3类白蚁中,圆唇凸额类白蚁和圆唇平额类白蚁亲缘关系相近,而两者与尖唇类白蚁亲缘关系较远。实验结果表明,我国存在异白蚁属,它与散白蚁属主要区别在于其表皮缺乏以下数种碳氢化合物：正十七烷烃、正二十烷烃、正二十一烷烃、正二十二烷烃、正二十三烷烃、正二十四烷烃和正二十六烷烃等;却含有一种特殊化合物异喹啉。表皮碳氢化合物分析的结果与形态分类的结果有一定差异,形态分类被鉴定为双色散白蚁的R. sp.1,被鉴定为小头散白蚁的R. sp.2,被鉴定为圆唇散白蚁的R.sp.3,被鉴定为尖唇散白蚁的H. sp.4,根据表皮碳氢化合物分析的结果,R. sp.1、R. sp.3和H. sp.4可能是其他种,而R. sp.2则可能是圆唇散白蚁的亚种或其他种。     

Genetic analysis of the breeding system of an invasive subterranean termite, Reticulitermes santonensis, in urban and natural habitats

Reticulitermes santonensis is a subterranean termite that invades urban areas in France and elsewhere where it causes damage to human-built structures. We investigated the breeding system, colony and population genetic structure, and mode of dispersal of two French populations of R. santonensis. Termite workers were sampled from 43 and 31 collection points, respectively, from a natural population in west-central France (in and around the island of Oleron) and an urban population (Paris). Ten to 20 workers per collection point were genotyped at nine variable microsatellite loci to determine colony identity and to infer colony breeding structure. There was a total of 26 colonies, some of which were spatially expansive, extending up to 320 linear metres. Altogether, the analysis of genotype distribution, F-statistics and relatedness coefficients suggested that all colonies were extended families headed by numerous neotenics (nonwinged precocious reproductives) probably descended from pairs of primary (winged) reproductives. Isolation by distance among collection points within two large colonies from both populations suggested spatially separated reproductive centres with restricted movement of workers and neotenics. There was a moderate level of genetic differentiation (F(ST) = 0.10) between the Oleron and Paris populations, and the number of alleles was significantly higher in Oleron than in Paris, as expected if the Paris population went through bottlenecks when it was introduced from western France. We hypothesize that the diverse and flexible breeding systems found in subterranean termites pre-adapt them to invade new or marginal habitats. Considering that R. santonensis may be an introduced population of the North American species R. flavipes, a breeding system consisting primarily of extended family colonies containing many neotenic reproductives may facilitate human-mediated spread and establishment of R. santonensis in urban areas with harsh climates.     

Cuticular hydrocarbons suggest three lineages in Reticulitermes (Isoptera: Rhinotermitidae) from North America

Cuticular hydrocarbon mixtures can be used to discriminate insect taxa. They have utility for determining phylogenetic relationships where they are independent characters with discrete states and represent a hierarchical distribution of shared, derived characters. We report inferred degrees of relatedness among the chemical phenotypes of Reticulitermes from PAUP (phylogenetic analysis using parsimony) analyses of cuticular hydrocarbon characters. One hundred and forty-one Reticulitermes colonies collected from California, Georgia, New Mexico, Arizona and Nevada were used. Initial maximum parsimony analyses sorted the 141 colonies into 26 chemical phenotypes. Subsequent analyses, using the ancestral species Coptotermes formosanus and Heterotermes sp. as outgroups, sorted Reticulitermes taxa into three major lineages, each characterized by a different set of dominant methyl-branched or unsaturated hydrocarbon components. Reticulitermes in lineage I have cuticular hydrocarbon mixtures with a preponderance of internally branched monomethylalkanes and 11,15-dimethylalkanes. Those in lineage II are defined by a preponderance of 5-methylalkanes and 5,17-dimethylalkanes. Taxa in lineage III are characterized by the predominance of olefins and a relative paucity of n-alkanes and methyl-branched alkanes. Bootstrap analyses and decay indices provided statistical support and robustness for these chemical-based relationships.     

Comparative study of the cuticular hydrocarbon composition of Melipona bicolor Lepeletier, 1836 (Hymenoptera, Meliponini) workers and queens

In social insects, cuticular hydrocarbons are involved in species, kin, caste and nestmate recognition. Gas chromatography and mass spectrometry were used to compare the cuticular hydrocarbon composition of workers, males and queens of Melipona bicolor. The cuticular hydrocarbon composition of this species was found to consist mainly of C23, C25:1, C25, C27:1, C27, C29:1 and C29, which are already present in imagoes that have not yet abandoned the brood cell. This composition varied quantitatively and qualitatively between and within the castes and sexes. The newly emerged workers and young queens (virgins) had similar cuticular hydrocarbon profiles, which were different from those of the males. When the females start executing their tasks in the colony, the cuticular hydrocarbon profile differences appear. The workers have less variety, while the queens conserve or increase the number of cuticular hydrocarbon compounds. The queens have more abdominal tegumentary glands than the workers, which apparently are the source of the new cuticular compounds.