Distinct chemical blends produced by different reproductive castes in the subterranean termite Reticulitermes flavipes

The production of royal pheromones by reproductives (queens and kings) enables social insect colonies to allocate individuals into reproductive and non-reproductive roles. In many termite species, nestmates can develop into neotenics when the primary king or queen dies, which then inhibit the production of additional reproductives. This suggests that primary reproductives and neotenics produce royal pheromones. The cuticular hydrocarbon heneicosane was identified as a royal pheromone in Reticulitermes flavipes neotenics. Here, we investigated the presence of this and other cuticular hydrocarbons in primary reproductives and neotenics of this species, and the ontogeny of their production in primary reproductives. Our results revealed that heneicosane was produced by most neotenics, raising the question of whether reproductive status may trigger its production. Neotenics produced six additional cuticular hydrocarbons absent from workers and nymphs. Remarkably, heneicosane and four of these compounds were absent in primary reproductives, and the other two compounds were present in lower quantities. Neotenics therefore have a distinct ‘royal’ blend from primary reproductives, and potentially over-signal their reproductive status. Our results suggest that primary reproductives and neotenics may face different social pressures. Future studies of these pressures should provide a more complete understanding of the mechanisms underlying social regulation in termites.

Endocuticle is involved in caste differentiation of the lower termite

Caste differentiation in termites is one of the most conspicuous examples of facultative polyphenism in animals. It is clear that specific cuticular formation occurs in hard exocuticles during caste differentiation. However, the developmental pattern of the soft endocuticle in the differentiation pathways of castes is unknown. To reveal whether the endocuticle is involved in caste differentiation, we compared the exocuticle and endocuticle thickness of individuals in 2 pathways (nymph line and worker line) of caste differentiation in the termite Reticulitermes aculabialis. The endocuticle protein genes were identified by transcriptome analysis and the expression patterns of these genes were confirmed in caste differentiation. We found that the endocuticle structure showed dynamic changes in 2 pathways, and the first difference in endocuticle structure occurred after larvae differentiation bifurcated into workers and nymphs. The thinning of the endocuticle was a significant event from nymphs developing into alates with the thickest exocuticle and thinnest endocuticle. The thickest endocuticle layers were found in the heads of the workers and the ultrastructure of the endocuticle in the heads was more complex than that in the thorax–abdomens. Six endocuticle protein genes were identified and annotated as endocuticle structural glycoproteins SgAbd-2, SgAbd-9, and Abd-5. The expression levels of endocuticle protein genes changed dramatically during caste development and the expression levels in neotenic reproductives (secondary reproductives) were significantly higher than those in alates (primary reproductives). These results reveal the roles of endocuticles in caste differentiation and adaptation to the environment.

A Glycoproteinaceous Secretion in the Seminal Vesicles of the Termite Coptotermes gestroi (Isoptera: Rhinotermitidae)

Coptotermes gestroi is a subterranean termite with colonies generally headed by a pair of primary reproductives, although neotenics may occur. In this study, the male reproductive system was compared during different life stages of nymphs, alates, neotenic reproductives, and kings of C. gestroi, focusing on the modifications of this system along the maturation of these individuals. The structure of the male reproductive system follows the pattern described for insects, although C. gestroi males do not exhibit conspicuous penises and differentiated accessory glands. In kings, each testis consisted of about seven lobes, significantly increased in size as compared to younger males. The spermatogenesis begins in third-instar nymphs, which already presented spermatozoa in the testes. The seminal vesicles are individualized in C. gestroi and have a secretory distal portion and a proximal portion with a role in spermatozoa storage. The secretion of the seminal vesicles is strongly periodic acid Schiff (PAS)-positive, whereas the xylidine Ponceau test revealed proteins that increase in quantity while the males become older. This is the first record of glycoproteins in the lumen of seminal vesicles in termites. Further studies will clarify how they are produced and interact in the physiology and nutrition of the non-flagellate spermatozoa of C. gestroi.

Diversity of Termite Breeding Systems

Termites are social insects that live in colonies headed by reproductive castes. The breeding system is defined by the number of reproductive individuals in a colony and the castes to which they belong. There is tremendous variation in the breeding system of termites both within and among species. The current state of our understanding of termite breeding systems is reviewed. Most termite colonies are founded by a primary (alate-derived) king and queen who mate and produce the other colony members. In some species, colonies continue throughout their life span as simple families headed by the original king and queen. In others, the primary king and queen are replaced by numerous neotenic (nymph- or worker-derived) reproductives, or less commonly primary reproductives, that are descendants of the original founding pair leading to inbreeding in the colony. In still others, colonies can have multiple unrelated reproductives due to either founding the colonies as groups or through colony fusion. More recently, parthenogenetic reproduction has shown to be important in some termite species and may be widespread. A major challenge in termite biology is to understand the ecological and evolutionary factors driving the variation in termite breeding systems.

Facultative asexual reproduction and genetic diversity of populations in the humivorous termite Cavitermes tuberosus

Termite colonies are typically founded by a pair of sexually reproducing dispersers, which can sometimes be replaced by some of their offspring. Some Reticulitermes and Embiratermes species routinely practice asexual queen succession (AQS): the queen is replaced by neotenic daughters produced by parthenogenesis, which mate with the primary king. Here, to cast light on the evolution of AQS, we investigated another candidate species, Cavitermes tuberosus (Termitinae). Of 95 nests, 39 contained a primary queen and 28 contained neotenic females (2–667 individuals), usually with the primary king. Microsatellite analyses confirmed that colonies were initiated by single pairs after large dispersal flights. More than 80% of the neotenic females were of exclusively maternal origin and completely homozygous, suggesting automictic parthenogenesis with gamete duplication. Conversely, workers, soldiers, and most alates and primary reproductives were produced sexually. AQS often occurs late, after colonies have reached maturity, whereas early AQS in other species may boost the young colony's growth rate. We suggest additional benefits of AQS in C. tuberosus , related with a smaller size, lesser stability and higher mobility of colonies. Our data add to the phylogenetical dispersion and diversity of modalities of AQS in termites, supporting a multiple evolutionary origin of this process.

Control of primary reproductives of Microtermes spp. in soil treated with Galleria cadavers infected with Heterorhabditis indica

The virulence of entomopathogenic nematodes (EPN) to the primary reproductives – alates and dealates – of Microtermes spp. was examined. Mortality in alates was observed in 18 h in the filter paper bioassay to test the relative virulence of infective juveniles (IJ) of Heterorhabditis indica and Steinernema abbasi. The LD50 for H. indica was achieved with 5.11 IJ alate−1 at 60 h, while for S. abbasi it was attained at 72 h with 6.91 IJ alate−1. Both nematodes completed their life cycle in the alates with an average production per cadaver of 3245 IJ and 2349 IJ for H. indica and S. abbasi, respectively, at an infection dose of 30 IJ alate−1. Thus H. indica was selected for the soil assay. The soil was implanted with H. indica-infected cadavers of Galleria mellonella. Within 72 h 100% mortality was observed in the dealates. Successful recycling of nematodes in the termite cadavers reinforced the suitability of H. indica for the biological control of primary reproductives.

Asexual queen succession in the subterranean termite Reticulitermes virginicus

Termite colonies are founded by a pair of primary reproductives. In many species, including subterranean termites (family Rhinotermitidae), the primary king and queen can be succeeded by neotenic reproductives that are produced from workers or nymphs within the colony. It is generally believed that these neotenics inbreed within the colony, sometimes for many generations. Here, we show that primary queens of the North American subterranean termite, Reticulitermes virginicus , are replaced by numerous parthenogenetically produced female neotenics. We collected functional female neotenics from five colonies of R. virginicus in North Carolina and Texas, USA. Genetic analysis at eight microsatellite loci showed that 91–100% of the neotenics present within a colony were homozygous at all loci, indicating that they were produced through automictic parthenogenesis with terminal fusion. In contrast, workers, soldiers and alates were almost exclusively sexually produced by mating between the female neotenics and a single king. This is the second termite species shown to undergo asexual queen succession, a system first described in the Japanese species, Reticulitermes speratus . Thus, the conditional use of sexual and asexual reproduction to produce members of different castes may be widespread within Reticulitermes and possibly other subterranean termites.

Disruption of the Termite Gut Microbiota and Its Prolonged Consequences for Fitness

ABSTRACTThe disruption of host-symbiont interactions through the use of antibiotics can help elucidate microbial functions that go beyond short-term nutritional value. Termite gut symbionts have been studied extensively, but little is known about their impact on the termite's reproductive output. Here we describe the effect that the antibiotic rifampin has not only on the gut microbial diversity but also on the longevity, fecundity, and weight of two termite species,Zootermopsis angusticollisandReticulitermes flavipes. We report three key findings: (i) the antibiotic rifampin, when fed to primary reproductives during the incipient stages of colony foundation, causes a permanent reduction in the diversity of gut bacteria and a transitory effect on the density of the protozoan community; (ii) rifampin treatment reduces oviposition rates of queens, translating into delayed colony growth and ultimately reduced colony fitness; and (iii) the initial dosages of rifampin had severe long-term fitness effects onZ. angusticollis. Taken together, our findings demonstrate that the antibiotic-induced perturbation of the microbial community is associated with prolonged reductions in longevity and fecundity. A causal relationship between these changes in the gut microbial population structures and fitness is suggested by the acquisition of opportunistic pathogens and incompetence of the termites to restore a pretreatment, native microbiota. Our results indicate that antibiotic treatment significantly alters the termite's microbiota, reproduction, colony establishment, and ultimately colony growth and development. We discuss the implications for antimicrobials as a new application to the control of termite pest species.