Termites can be a significant source of N2O emissions in tropical and subtropical ecosystems. The emission rates vary greatly between species, with many species creating emission hotspots while others acting as net sinks. We studied the relationships between net and gross N2O production/consumption and the abundances of eight nitrifier and denitrifier groups (as determined by functional marker genes) in termite gut homogenates for eleven species from five feeding guilds. Overall, the abundance of nitrite reducers and, to a lesser extent, nitrifiers in the gut was strongly correlated with gross N2O production, whereas N2O reduction was correlated with the abundance of nosZ1 N2O reducers and the nosZ1/nirK ratio. Our results show that the differences in gross and net N2O production rates between termite species are primarily due to differences in nitrifier and denitrifier abundances, as well as the ratio of related functional gene marker abundances in the guts. N2O production rates were negatively correlated to the ratio of nosZ to nir. N2O production rates by live individuals measured for a subset of termite species were lower in the wood-feeding than in other species. Our results suggest that the differences in functional gene abundances may be associated with feeding guild, although this should be tested on a larger number of termite species.

Kings and queens of eusocial termites can live for decades, while queens sustain a nearly maximal fertility. To investigate the molecular mechanisms underlying their long lifespan, we carried out transcriptomics, lipidomics and metabolomics in Macrotermes natalensis on sterile short-lived workers, long-lived kings and five stages spanning twenty years of adult queen maturation. Reproductives share gene expression differences from workers in agreement with a reduction of several aging-related processes, involving upregulation of DNA damage repair and mitochondrial functions. Anti-oxidant gene expression is downregulated, while peroxidability of membranes in queens decreases. Against expectations, we observed an upregulated gene expression in fat bodies of reproductives of several components of the IIS pathway, including an insulin-like peptide, Ilp9 . This pattern does not lead to deleterious fat storage in physogastric queens, while simple sugars dominate in their hemolymph and large amounts of resources are allocated towards oogenesis. Our findings support the notion that all processes causing aging need to be addressed simultaneously in order to prevent it.

Eusocial termite queens achieve nearly maximal fertility throughout their extremely long life without apparent signs of aging. Termites represent, therefore, an ideal model for aging research. To investigate the molecular mechanisms underlying their long reproductive life, we carried out transcriptomic, lipidomic and metabolomic analyses on fat bodies of sterile short lived workers, long-lived kings and five stages spanning twenty years of adult queen maturation. In mature reproductives, genes supporting a robust mitochondrial functioning or associated with genome stability were upregulated. In most organisms, insulin signaling increases fertility but decreases lifespan, often accompanied by harmful lipid signatures. Our findings suggest that an upregulation of insulin-like peptide (Ilp9) in the fat body of termite queens is accompanied by a specific lipid metabolism, limiting fat storage, thus sustaining both high fertility and maintaining extreme lifespan. Our results highlight potential molecular targets for research into aging-related metabolic diseases linked to the accumulation of excess fat.

Background Termites account for natural biomass utilization systems (NBUS) that evolved the ability to overcome the overall recalcitrance of lignins towards lignocellulose transformation processes. With the objective of applying this capacity to the conversion of technical lignins produced by biorefineries, a higher wood-feeding termite species, Nasutitermes ephratae was fed with a commercial grass soda lignin (Protobind 1000, PB1000). The survival rates of Protobind 1000-fed termites were determined as well as changes in the structure of gut bacterial community and in the chemical composition of this technical lignin. Results The ingestion of PB1000 by worker castes of N. ephratae was revealed by Pyrolysis-Gas Chromatography Mass Spectrometry (Py-GC/MS) analyses directly performed on termites. Survival rates were reduced by two –fold in the termites fed with PB1000 compared to controls. The relative abundance of Firmicutes and Bacteroidetes increased in the gut bacterial community of termites fed with PB1000. The digestion of PB1000 by termites triggered an increase in the syringyl-to-guaiacyl (S/G) ratios. These changes in the chemical composition of PB1000 in the gut of termites was marked by a decrease in relative content of free phenolic monomers. Conclusion This work showed the abilities of digestive tract of a wood-feeding higher termite species, N. ephratae to metabolize the fraction of the volatile phenolic monomers of PB1000. Overall, our results provide insights into the bacterial lineage candidates for development of bacterial inoculum for pretreatment processes in valorization of technical lignin in biorefinery.

Congeneric species that live in sympatry may have evolved various mechanisms that maintain reproductive isolation among species. However, with the spread of invasive organisms owing to increased global human activity, some species that evolved in allopatry can now be found outside their native range and may have the opportunity to interact, in the absence of mechanisms for reproductive isolation. In South Florida, where the Asian subterranean termite, Coptotermes gestroi (Wamann), and the Formosan subterranean termite, Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) are invasive, the two species can engage in heterospecific mating behavior as their distribution range and their dispersal flight season both overlap. Termites rely on semiochemicals for many of their activities, including finding a mate after a dispersal flight. In this study, we showed that females of both species produce (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol (DTE) from their tergal glands as a shared sex pheromone. We suggest that both species primarily rely on an inundative dispersal flight strategy to find a mate, and that DTE is used as a short distance pheromone or contact pheromone to initiate and maintain the tandem between males and females. The preference of C. gestroi males for C. formosanus females during tandem resulted from the relatively high amount of DTE produced by tergal glands of C. formosanus females, when compared with those of C. gestroi females. This results in confusion of mating in the field during simultaneous dispersal flights, with a potential for hybridization. Such observations imply that no prezygotic barriers emerged while the two species evolved in allopatry for ~18 Ma.

Trail-following behavior is a key to ecological success of termites, allowing them to orient themselves between the nesting and foraging sites. This behavior is controlled by specific trail-following pheromones produced by the abdominal sternal gland occurring in all termite species and developmental stages. Trail-following communication has been studied in a broad spectrum of species, but the “higher” termites (i.e. Termitidae) from the subfamily Syntermitinae remain surprisingly neglected. To fill this gap, we studied the trail-following pheromone in six genera and nine species of Syntermitinae. Our chemical and behavioral experiments showed that (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol is the single component of the pheromone of all the termite species studied, except for Silvestritermes euamignathus. This species produces both (3Z,6Z)-dodeca-3,6-dien-1-ol and neocembrene, but only (3Z,6Z)-dodeca-3,6-dien-1-ol elicits trail-following behavior. Our results indicate the importance of (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol, the most widespread communication compound in termites, but also the repeated switches to other common pheromones as exemplified by S. euamignathus.

There are multiple forms of interactions between termites and bacteria. In addition to their gut microbiota, which has been intensively studied, termites host intracellular symbionts such as Wolbachia. These distinct symbioses have been so far approached independently and mostly in adult termites. We addressed the dynamics of Wolbachia and the microbiota of the eggs and gut for various life stages and castes of the wood-feeding termite, Nasutitermes arborum, using deep-sequencing of the 16S rRNA gene. Wolbachia was dominant in eggs as expected. Unexpectedly, it persisted in the gut of nearly all stages and castes, indicating a wide somatic distribution in termites. Wolbachia-related sequences clustered into few operational taxonomic units, but these were within the same genotype, acquired maternally. Wolbachia was largely dominant in DNA extracts from the guts of larvae and pre-soldiers (59.1%-99.1% of reads) where gut-resident lineages were less represented and less diverse. The reverse was true for the adult castes. This is the first study reporting the age-dependency of the relative abundance of Wolbachia in the termite gut and its negative correlation with the diversity of the microbiota. The possible mechanisms underlying this negative interaction are discussed.

Studies on termite symbiosis have revealed that significant symbiont lineages are maintained across generations. However, most studies have focused only on the worker caste. Little is known about the gut microbiota of reproductives, the most probable vectors for transmitting these lineages to offspring. Using 16S rRNA gene-based Illumina MiSeq sequencing, we compared the gut microbiota of swarming alates of the higher termite Nasutitermes arborum with those of their nestmates from the parental colony. The OTU-based alpha diversity indices showed that the gut microbiota of the alates was at least as diverse as those of non-reproductive adults. It was largely dominated by Spirochaetes mostly of the Treponema I cluster (63.1% of reads), the same dominant taxa found in soldiers and workers of this species and in workers of closely related Nasutitermes species. The termite-specific lineages also included other representative taxa such as several clusters of Bacteroidetes and Fibrobacteres-TG3 group. The microbiota of alates was dominated by a core set of host-specific lineages (87% of reads, 77.6% of OTUs), which were always present across all castes/stages. This first comprehensive survey of the microbiota of the founding reproductives of these xylophagous higher termites shows that the bulk of the host endogenous symbionts, mostly taxa that cannot thrive outside the gut, are brought from the parent colony. The royal pair therefore seems to be a key player in the transmission of symbionts across generations and thereby in host-symbiont codiversification. The high proportion of fiber-degrading lineages in their gut suggests a wood-rich diet unlike the larval stages.

Coptotermes Wasmann (Isoptera: Rhinotermitidae) is one of the most economically important subterranean termite genera and some species are successful invaders. However, despite its important pest status, the taxonomic validity of many named Coptotermes species remains unclear. In this study, we reviewed all named species within the genus and investigated evidence supporting the validity of each named species. Species were systematically scrutinized according to the region of their original description: Southeast Asia, India, China, Africa, the Neotropics, and Australia. We estimate that of the currently 69 named species described by accepted nomenclatural rules, only 21 taxa have solid evidence for validity, 44 names have uncertain status, and the remaining species names should be synonymized or were made unavailable. Species with high degrees of invasiveness may be known under additional junior synonyms due to independent parochial descriptions. Molecular data for a vast majority of species are scarce and significant effort is needed to complete the taxonomic and phylogenetic revision of the genus. Because of the wide distribution of Coptotermes, we advocate for an integrative taxonomic effort to establish the distribution of each putative species, provide specimens and corresponding molecular data, check original descriptions and type specimens (if available), and provide evidence for a more robust phylogenetic position of each species. This study embodies both consensus and contention of those studying Coptotermes and thus pinpoints the current uncertainty of many species. This project is intended to be a roadmap for identifying those Coptotermes species names that need to be more thoroughly investigated, as an incentive to complete a necessary revision process.

In the tropics, termites are major players in the mineralization of organic matter leading to the production of greenhouse gases including nitrous oxide (N2O). Termites have a wide trophic diversity and their N-metabolism depends on the feeding guild. This study assessed the extent to which N2O emission levels were determined by termite feeding guild and tested the hypothesis that termite species feeding on a diet rich in N emit higher levels of N2O than those feeding on a diet low in N. An in-vitro incubation approach was used to determine the levels of N2O production in 14 termite species belonging to different feeding guilds, collected from a wide range of biomes. Fungus-growing and soil-feeding termites emit N2O. The N2O production levels varied considerably, ranging from 13.14 to 117.62 ng N2O-N d-1 (g dry wt.)-1 for soil-feeding species, with Cubitermes spp. having the highest production levels, and from 39.61 to 65.61 ng N2O-N d-1 (g dry wt.)-1 for fungus-growing species. Wood-feeding termites were net N2O consumers rather than N2O producers with a consumption ranging from 16.09 to 45.22 ng N2O-N d-1 (g dry wt.)-1. Incubating live termites together with their mound increased the levels of N2O production by between 6 and 13 fold for soil-feeders, with the highest increase in Capritermes capricornis, and between 14 and 34 fold for fungus-growers, with the highest increase in Macrotermes muelleri. Ammonia-oxidizing (amoA-AOB and amoA-AOA) and denitrifying (nirK, nirS, nosZ) gene markers were detected in the guts of all termite species studied. No correlation was found between the abundance of these marker genes and the levels of N2O production from different feeding guilds. Overall, these results support the hypothesis that N2O production rates were higher in termites feeding on substrates with higher N content, such as soil and fungi, compared to those feeding on N-poor wood.

Termites have developed many exocrine glands, generally dedicated to defence or communication. Although a few of these glands occur in all termite species, or represent synapomorphies of larger clades, others are morphological innovations of a single species, or a few related species. Here, we describe the nasus gland, a new gland occurring at the base of the nasus of Angularitermes soldiers. The nasus gland is composed of class 1, 2, and 3 secretory cells, a rare combination that is only shared by the sternal and tergal glands of some termites and cockroaches. The ultrastructural observations suggest that the secretion is produced by class 2 and 3 secretory cells, and released mostly by class 3 cells. The base of the nasus has a rough appearance due to numerous pits bearing openings of canals conducting the secretion from class 3 secretory cells to the exterior. We tentatively assign a defensive function to the nasus gland, although further research is needed to confirm this function. Although the gland is described only from species of Angularitermes, other genera of Nasutitermitinae also present a rough nasus base, suggesting the presence of a similar, possibly homologous, gland.

Pseudacanthotermes spiniger and P. militaris are two African fungus-growing termites (Termitidae, Macrotermitinae) which may become pests in disturbed agrosystems where they often live in sympatry. To study their development and their reproductive strategies, colonies of both species were reared in the laboratory for 20 and 17 years, respectively, after their foundation from reproductive pairs. The first steps of development were in great part similar in both species, although P. spiniger favoured the defence during the juvenile period, while P. militaris tended to favour a rapid development. While P. spiniger colonies did not produce alates until year 7 of colony life, P. militaris colonies were able to produce a fertile progeny 4 years after their foundation. In contrast, major soldiers were more rapidly differentiated in the incipient colonies of P. spiniger. Dispersal flights occurred every year for 10 years in P. spiniger. In P. militaris, dispersal flights did not occur regularly although alates appeared yearly. The annual number of alates produced by P. spiniger increased with the colony age to reach a maximum of 25,000 individuals and global production of alates was estimated at ca. 150,000 individuals in the life of a colony. The longevity of P. spiniger and P. militaris colonies was around 20 years. These species were shown to be reproductively isolated by multiple pre-mating mechanisms. While chronological differences in dispersal flights contribute to reproductive isolation of the two species, the non-viability of experimental hybrid colonies also indicates the involvement of post-mating mechanisms of isolation.

* INRA, UR50, Laboratoire de Biotechnologie de l’Environnement, Avenue des Etangs, Narbonne, F11100, France (E-mail: godon@supagro.inra.fr; escudie@supagro.inra.f r ; audrey.battimelli@supagro.inra.fr ; torrijos@supagro.inra.fr; steyer@supagro.inra.fr) **CEMAGREF-HBAN, Parc de Tourvoie BP 44, F-92163 Antony cedex, France (E-mail: ariane.bize@cemagref.fr) *** INRA, UMR1319, MICALIS, PAPPSO, Domaine de Vilvert, F-78352 Jouy-en-Josas cedex, France (E-mail: alain.guillot@jouy.inra.fr) **** Suez Environnement, 38 Rue du president Wilson , F-78230 Le Pecq, France (E-mail: Tianlun.LI@suez-env.com) ***** UMR 211 BIOEMCO-IBIOS, UPEC, 61 Avenue du General de Gaulle, Creteil F-94010, France (E-mail: miambi@univ-paris12.fr) ****** IRD, UMR 211 BIOEMCO-IBIOS, 32 Avenue Henri Varagnat, Bondy, F93143, France (E-mail: Alain.Robert@bondy.ird.fr)

To provide a better understanding of soil–atmosphere gas exchange processes, this study describes the atmospheric nitrous oxide (N2O) uptake by xylophagous termites and the biological process involved. The N2O consumption rates of three xylophagous termite species (Hodotermes mossambicus, Nasutitermes voeltzkowi and Hodotermopsis sjoestedti) were determined in incubation vials with ambient, artificially enhanced N2O concentrations in the headspace. Live individuals of the three termite species significantly decreased N2O concentrations (88%) in the headspace of the vials after 24 h incubation in the dark. The acetylene reduction assay method applied to N. voeltzkowi, a xylophagous termite species, showed a decrease in N2O uptake in acetylene-treated individuals, indicating the potential involvement of termite gut denitrifying microbes. The N2 formed is potentially subjected to assimilation via nitrogenase reductase into termite biomass through biological fixation as demonstrated by the reduction of acetylene to ethylene at an average rate of 18.21 ± 1.34 nmol C2H4 g−1 dw d−1. Further studies should focus on measurements of N2O-reductase (nosZ) gene activity in termite guts to gain a better understanding of the N2O reduction process in xylophagous termite species.

In order to maintain healthy colonies, termite workers dispose of the cadavers of dead nest mates by cannibalism, burial, or necrophoresis. However, when multiple reproductives found a new colony by pleometrosis, there are no worker castes at the early stages of the foundation to eliminate or isolate the corpses. In this study, we showed that in young pleometrotic colonies, reproductives of Pseudacanthotermes spiniger had the ability to perform this task. Because of the claustral conditions, and the potential inability of the dealates to feed on their own, their behaviour was restricted to the burial of the cadaver within the initial chamber. This burial behaviour, previously not reported in the reproductive caste of termites, appeared to be induced by chemical signals released by the corpses during decomposition, among which various fatty acids, indole and phenol were the most active. The burial finally resulted in the physical isolation of corpses, thus reducing the chances for opportunistic pathogens to spread among the rest of the individuals.

The trail-following pheromone and sex pheromones were investigated in the Indomalayan termite Hodotermopsis sjoestedti belonging to the new family Archotermopsidae. Gas chromatography coupled to mass spectrometry (GC-MS) after solid phase microextraction (SPME) of the sternal gland secretion of pseudergates and trail-following bioassays demonstrated that the trail-following pheromone of H. sjoestedti was syn-4,6-dimethylundecan-1-ol, a new chemical structure for termite pheromones. GC-MS after SPME of the sternal gland secretion of alates also allowed the identification of sex-specific compounds. In female alates, the major sex-specific compound was identified as (5E)-2,6,10-trimethylundeca-5,9-dienal, a compound previously identified as the female sex pheromone of the termite Zootermopsis nevadensis. In male alates, the major sex-specific compound was identified as syn-4,6-dimethylundecanal, a homolog of syn-4,6-dimethyldodecanal, which has previously been confirmed as the male sex pheromone of Z. nevadensis. The presence of sex-specific compounds in alates of H. sjoestedti strongly suggests for this termite the presence of sex-specific pairing pheromones which were only known until now in Z. nevadensis. Our results showed therefore a close chemical relationship between the pheromones of the taxa Hodotermopsis and Zootermopsis and, in contrast, a clear difference with the taxa Stolotermes and Porotermes, which is in total agreement with the recent creation of the families Archotermopsidae and Stolotermitidae as a substitute for the former family Termopsidae.

Termites (Isoptera) are eusocial insects, which live in an environment that can favor the spread of pathogens. To reduce the chance of an epizootic within a colony, termites have evolved many defense mechanisms. Most studies have focused on the social aspect of disease resistance, while the individual capacity of a termite to survive an infection remains poorly documented. We previously showed that when the eastern subterranean termite, Reticulitermes flavipes (Kollar), was exposed to the entomopathogenic fungus, Metarhizium anisopliae (Metch.) Sorokin, cellular encapsulation of the penetrating fungus was one of the last lines of defense for individual termites to prevent internal mycosis. The current study used histological preparations to (i) compare cellular encapsulation of M. anisopliae among 6 termite species from 5 families that evolved in habitats with different pathogenic pressures, and (ii) examine the effect of cellular encapsulation on the survival of termites exposed to M. anisopliae. Our results showed that all termite species were able to use hemocytes to encapsulate M. anisopliae when this fungus penetrated through the insect cuticle, but that the physiological cost to successfully encapsulate M. anisopliae varied greatly among termite species. We suggest that termite species, which evolved in a habitat with high pathogenic pressure, are adapted with more efficient immune reactions than those that evolved in a habitat with low pathogenic pressure.

Amitermes evuncifer (termitidae, termitinae) is a pest termite in several countries of Africa, damaging crops and buildings. The trail-following pheromone of this termite was isolated from worker sternal glands identified by GC-MS after solid phase microextraction (SPME). The pheromone is composed of (3Z,6Z,8E)-dodeca-3,6,8-trien-1-ol (dodecatrienol) and neocembrene acting in synergy, neocembrene being 5-10 times more abundant than dodecatrienol. Dodecatrienol alone is able to elicit trail-following in workers, whereas neocembrene alone is inactive. However, the association of both components induces a significantly higher trail-following activity. Multicomponent trail-following pheromones were only known, until now, in the termitidae Nasutitermitinae, and in the Rhinotermitidae Prorhinotermes.

In the present study, trail pheromone blends are identified for the first time in termites. In the phylogenetically complex Nasutitermitinae, trail-following pheromones are composed of dodecatrienol and neocembrene, the proportions of which vary according to species, although neocembrene is always more abundant than dodecatrienol (by 25-250-fold). Depending on species, termites were more sensitive to dodecatrienol or to neocembrene but the association of both components always elicited significantly higher trail following, with a clear synergistic effect in most of the studied species. A third component, trinervitatriene, was identified in the sternal gland secretion of several species, but its function remains unknown. The secretion of trail pheromone blends appears to be an important step in the evolution of chemical communication in termites. The pheromone optimizes foraging, and promotes their ecological success. (C) 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99, 20-27.