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Colony genetic organization and colony fusion in the termite Reticulitermes flavipes as revealed by foraging patterns over time and space.

Temporal and spatial analyses are seldom utilized in the study of colony genetic structure, but they are potentially powerful methods which can yield novel insights into the mechanisms underlying variation in breeding systems. Here we present the results of a study which incorporated both of these dimensions in an examination of genetic structure of subterranean termites in the genus Reticulitermes (primarily R. flavipes). Most colonies of this species (70%) were simple families apparently headed by outbred primary reproductives, while most of the remaining (27% of the total) colonies contained low effective numbers of moderately inbred reproductives. Mapping the spatial distribution of colony foraging sites over time revealed that despite the high colony density, the absolute foraging boundaries of most R. flavipes colonies were persistent and exclusive of other conspecific colonies, which suggests that this species is more territorial than has been implied by laboratory studies of intraspecific aggression. Nevertheless, we found a single colony (3% of all colonies) which contained the offspring of more than two unrelated reproductives. Although other studies have also described subterranean termite colonies with a similarly complex genetic composition, we demonstrate here that such colonies can form under natural conditions via the fusion of whole colonies. This study underscores how repeated sampling from individual colonies over time and space can yield information about colony spatial and genetic structure that cannot be obtained from conventional analyses or sampling methods.

Animals↗

On the origin of termite workers: weighing up the phylogenetic evidence.

Resolving the phylogenetic history of a 'true' worker caste in termites is essential to our understanding of termite eusocial evolution. Whether this caste is ancient and monophyletic or derived and polyphyletic will have a tremendous impact on our interpretation of termite eusocial history and remains an outstanding question in termite biology. Recent work has begun to re-examine this question in light of new phylogenetic information, but new questions have now arisen about how best to model character state changes in termite caste systems. In the present paper, we compare the models of Grandcolas and D'Haese [J. Evol. Biol. 15 (2002) 885] and Thompson et al. [J. Evol Biol. 13 (2000) 8691 and attempt to make explicit how these proposals differ with respect to the number of, and homology between, character states. We highlight the support each model has for the two principal, but competing, evolutionary hypotheses outlined above.

Animals↗

The origin of a 'true' worker caste in termites: mapping the real world on the phylogenetic tree.

The evolution of the 'true' worker caste in termites is not decisively inferred by coding and mapping both this character and the foraging behaviour on a phylogenetic tree. Answering to Thompson et al. (2000, 2003), and with reference to Grandcolas and D'Haese (2002), we show that this indecisive inference depends on the correct consideration paid to the outgroups. These last ones could be non subsocial cockroaches, or some wood-eating subsocial cockroaches often considered misleadingly as living ancestors, or even any hemimetabolous insects, all of them would be unambiguously lacking 'true' worker caste and pseudergate caste and not showing the 'one-piece' life type foraging behaviour. These statements derive from observing, coding and mapping the real world on the tree without making ad hoc assumptions. In that respect, because termites do not exist in isolation, apart from the tree of life, mapping the character of interest on the tree must be applied to the outgroups as well.

Animals↗

Phylogenetic diversity of termite gut spirochaetes.

A molecular phylogenetic analysis was done of not-yet-cultured spirochaetes inhabiting the gut of the termite, Reticulitermes flavipes (Kollar). Ninety-eight clones of near-full-length spirochaetal 16S rDNA genes were classified by ARDRA pattern and by partial sequencing. All clones grouped within the genus Treponema, and at least 21 new species of Treponema were recognized within R. flavipes alone. Analysis of 190 additional clones from guts of Coptotermes formosanus Shiraki and Zootermopsis angusticollis (Hagen), as well as published data on clones from Cryptotermes domesticus (Haviland), Mastotermes darwiniensis Froggatt, Nasutitermes lujae (Wasmann) and Reticulitermes speratus(Kolbe), revealed a similar level of novel treponemal phylogenetic diversity in these representatives of five of the seven termite families. None of the clones was closely related (i.e. all bore < or = 91% sequence similarity) to any previously recognized treponeme. The data also revealed the existence of two major phylogenetic groups of treponemes: one containing all of the currently known isolates of Treponema and a large number of phylotypes from the human gingival crevice, but only a minority of the termite gut spirochaete clones; another containing the majority of termite spirochaete clones and two Spirochaeta (S. caldaria and S. stenostrepta), which, although free living, group within the genus Treponema on the basis of 16S rRNA sequence. Signature nucleotides that almost perfectly distinguished the latter group, herein referred to as the 'termite cluster', occurred at the following (E. coli numbering) positions: 289-G x C-311; A at 812; and an inserted nucleotide at 1273. The emerging picture is that the long-recognized and striking morphological diversity of termite gut spirochaetes is paralleled by their phylogenetic diversity and may reflect substantial physiological diversity as well.

Animals↗

Impact of oxygen on metabolic fluxes and in situ rates of reductive acetogenesis in the hindgut of the wood-feeding termite Reticulitermes flavipes.

The symbiotic digestion of lignocellulose in the hindgut of the wood-feeding termite Reticulitermes flavipes is characterized by two major metabolic pathways: (i) the oxidation of polysaccharides to acetate by anaerobic hydrogen-producing protozoa; and (ii) the reduction of CO2 by hydrogenotrophic acetogenic bacteria. Both reactions together would render the hindgut largely homoacetogenic. However, the results of this study show that the situation is more complex. By microinjection of radiolabelled metabolites into intact agarose-embedded hindguts, we showed that the in situ rates of reductive acetogenesis (3.3 nmol termite(-1) h(-1)) represent only 10% of the total carbon flux in the living termite, whereas 30% of the carbon flux proceeds via lactate. The rapid turnover of the lactate pool (7.2 nmol termite(-1) h(-1)) consolidates the previously reported presence of lactic acid bacteria in the R. flavipes hindgut and the low lactate concentrations in the hindgut fluid. However, the immediate precursor of lactate remains unknown; the low turnover rates of injected glucose (< 0.5 nmol termite(-1) h(-1)) indicate that free glucose is not an important intermediate under in situ conditions. The influence of the incubation atmosphere on the turnover rate and the product pattern of glucose and lactate confirmed that the influx of oxygen via the gut epithelium and its reduction in the hindgut periphery have a significant impact on carbon and electron flow within the hindgut microbial community. The in situ rates of reductive acetogenesis were not significantly affected by the presence of oxygen or exogenous H2, which is in agreement with a localization of homoacetogens in the anoxic gut lumen rather than in the oxic periphery. This adds strong support to the hypothesis that the co-existence of methanogens and homoacetogens in this termite is based on the spatial arrangement of the different populations of the gut microbiota. A refined model of metabolic fluxes in the hindgut of R. flavipes is presented.

Acetates↗

Spirochete and protist symbionts of a termite (Mastotermes electrodominicus) in Miocene amber.

Extraordinary preservation in amber of the Miocene termite Mastotermes electrodominicus has led to the discovery of fossil symbiotic microbes. Spirochete bacteria and wood-digesting protists were identified in the intestinal tissue of the insect. Fossil wood (xylem: developing vessel-element cells, fibers, pit connections), protists (most likely xylophagic amitochondriates), an endospore (probably of the filamentous intestinal bacterium Arthromitus = Bacillus), and large spirochetes were seen in thin section by light and transmission electron microscopy. The intestinal microbiota of the living termite Mastotermes darwiniensis, a genus now restricted to northern Australia, markedly resembles that preserved in amber. This is a direct observation of a 20-million-year-old xylophagus termite fossil microbial community.

Amber↗

Termites assess wood size by using vibration signals.

Contrary to the common perception that termites are indiscriminant eaters, termites choose their food carefully; however, the methods by which they choose food are not well understood. Using choice experiments and recordings of termites feeding on wooden blocks of different sizes, we show that worker drywood termites (Cryptotermes domesticus) use the resonant frequency of a block of wood to assess its size. Drywood termites showed differences in their response to vibration recordings of termites compared with artificially generated signals, suggesting that they can discriminate the source of vibration. Furthermore, fewer workers matured into neotenic reproductives when recorded termite signals were played, suggesting that vibration signals play an important role in termite communication.

Animals↗

Social exploitation of hexamerin: RNAi reveals a major caste-regulatory factor in termites.

Lower termites express a unique form of eusocial polyphenism in that totipotent workers can differentiate into either soldier or reproductive caste phenotypes. In this initial effort using RNA interference in termites, we found that two hexamerin genes, Hex-1 and Hex-2, participate in the regulation of caste polyphenism. Our methodology involved a dual gene-silencing approach that used a single short-interfering RNA fragment to silence the two homologous hexamerin genes. We performed validation studies that evaluated effects on nontarget housekeeping genes, silencing of a nonhousekeeping control gene, and effects at the protein level. We found that the two hexamerin proteins, which are inducible by the morphogenetic juvenile hormone and which constitute a significant proportion of total termite protein, suppress juvenile-hormone-dependent worker differentiation to the soldier caste phenotype. This mechanism allows termite colonies to retain high proportions of altruistic worker caste members, thus apparently enhancing colony-inclusive fitness. These findings demonstrate a unique status quo regulatory mechanism for termite worker caste retention and provide an example of previously undescribed preadult developmental/caste-regulatory genes from any social insect.

Amino Acid Sequence↗

The development of immunity in a social insect: evidence for the group facilitation of disease resistance.

The extraordinary diversity and ecological success of the social insects has been attributed to their ability to cope with the rich and often infectious microbial community inhabiting their nests and feeding sites. Mechanisms of disease control used by eusocial species include antibiotic glandular secretions, mutual grooming, removal of diseased individuals from the nest, and the innate and adaptive immune responses of colony members. Here we demonstrate that after a challenge exposure to the entomopathogenic fungus Metarhizium anisopliae, dampwood termites Zootermopsis angusticollis have higher survivorship when individuals develop immunity as group members. Furthermore, termites significantly improve their ability to resist infection when they are placed in contact with previously immunized nestmates. This "social transfer" of infection resistance, a previously unrecognized mechanism of disease control in the social insects, could explain how group living may improve the survivorship of colony members despite the increased risks of pathogen transmission that can accompany sociality.

Animals↗

Evolution of eusociality and the soldier caste in termites: influence of intraspecific competition and accelerated inheritance.

We present new hypotheses and report experimental evidence for powerful selective forces impelling the evolution of both eusociality and the soldier caste in termites. Termite ancestors likely had a nesting and developmental life history similar to that of the living family Termopsidae, in which foraging does not occur outside the host wood, and nonsoldier helpers retain lifelong options for differentiation into reproductives. A local neighborhood of families that live exclusively within a limited resource results in interactions between conspecific colonies, high mortality of founding reproductives, and opportunities for accelerated inheritance of the nest and population by offspring that differentiate into nondispersing neotenic reproductives. In addition, fertile reproductive soldiers, a type of neotenic previously considered rare and docile, frequently develop in this intraspecific competitive context. They can be highly aggressive in subsequent interactions, supporting the hypothesis that intercolonial battles influenced the evolution of modern sterile termite soldier weaponry and behaviors.

Animals↗

The evolution of fungus-growing termites and their mutualistic fungal symbionts.

We have estimated phylogenies of fungus-growing termites and their associated mutualistic fungi of the genus Termitomyces using Bayesian analyses of DNA sequences. Our study shows that the symbiosis has a single African origin and that secondary domestication of other fungi or reversal of mutualistic fungi to a free-living state has not occurred. Host switching has been frequent, especially at the lower taxonomic levels, and nests of single termite species can have different symbionts. Data are consistent with horizontal transmission of fungal symbionts in both the ancestral state of the mutualism and most of the extant taxa. Clonal vertical transmission of fungi, previously shown to be common in the genus Microtermes (via females) and in the species Macrotermes bellicosus (via males) [Johnson, R. A., Thomas, R. J., Wood, T. G. & Swift, M. J. (1981) J. Nat. Hist. 15, 751-756], is derived with two independent origins. Despite repeated host switching, statistical tests taking phylogenetic uncertainty into account show a significant congruence between the termite and fungal phylogenies, because mutualistic interactions at higher taxonomic levels show considerable specificity. We identify common characteristics of fungus-farming evolution in termites and ants, which apply despite the major differences between these two insect agricultural systems. We hypothesize that biparental colony founding may have constrained the evolution of vertical symbiont transmission in termites but not in ants where males die after mating.

Animals↗

Evidence of termite foraging by Swartkrans early hominids.

Previous studies have suggested that modified bones from the Lower Paleolithic sites of Swartkrans and Sterkfontein in South Africa represent the oldest known bone tools and that they were used by Australopithecus robustus to dig up tubers. Macroscopic and microscopic analysis of the wear patterns on the purported bone tools, pseudo bone tools produced naturally by known taphonomic processes, and experimentally used bone tools confirm the anthropic origin of the modifications. However, our analysis suggests that these tools were used to dig into termite mounds, rather than to dig for tubers. This result indicates that early hominids from southern Africa maintained a behavioral pattern involving a bone tool material culture that may have persisted for a long period and strongly supports the role of insectivory in the early hominid diet.

Animals↗

Trichosporon mycotoxinivorans sp. nov., a new yeast species useful in biological detoxification of various mycotoxins.

A yeast strain isolated from the hindgut of the lower termite Mastotermes darwiniensis (Mastotermitidae) was found to represent a new member of the genus Trichosporon. Trichosporon mycotoxinivorans is closely related to T. loubieri on the basis of the phylogenetic trees based on the D1/D2 region of 26S rDNA, an approx. 600 bp fragment of the 18S rDNA and both ITS regions. However, the two species differ at nine positions in the D1/D2 region of 26S rDNA. The IGS1 region of T. mycotoxinivorans is 401 bp long. T. mycotoxinivorans is distinguished from T. loubieri by its ability to assimilate inulin and galactitol, and its inability to grow at 40 degrees C. The name of this newly isolated strain refers to an important characteristics of T. mycotoxinivorans to detoxify mycotoxins such as ochratoxin A and zearalenone. Therefore this strain can be used for the deactivation of the respective mycotoxins in animal feeds.

Animals↗

Phylogenetic diversity and whole-cell hybridization of oxymonad flagellates from the hindgut of the wood-feeding lower termite Reticulitermes flavipes.

SSU rRNA genes of oxymonad protists from the hindgut of the wood-feeding termite Reticulitermes flavipes were PCR-amplified using a newly designed oxymonad-specific forward primer and a newly designed reverse primer specific for termite gut flagellates. After cloning, the clone library was sorted into four groups by RFLP analysis and nearly full-length SSU rRNA gene sequences were obtained for representative clones from each group. Phylogenetic analysis revealed that sequences of all four groups formed a monophyletic cluster with the only other existing SSU rRNA gene sequence of oxymonads. Using whole-cell hybridization with clone-specific fluorescently labeled probes, each of the four clone groups could be assigned to a specific morphotype, which were identified as Dinenympha gracilis, Dinenympha fimbriata, and so-far undescribed species of Pyrsonympha and Dinenympha. Our results demonstrate that the morphological variety of oxymonads is not caused by the presence of different developmental stages of the same organism, but that the various morphotypes represent different species.

Animals↗

Biological and chemical assays of pyrethroids in cattle dung.

Bioassays were developed in Zimbabwe to measure pyrethroid in cattle dung. These and chemical assays then estimated concentrations in dung from treated oxen and elucidated risks to dung fauna. Laboratory bioassays with adult beetles (Histeridae and Scarabaeinae, including Copris, Digitonthophagus, Onitis and Sisyphus spp.) and muscoid larvae (Musca lusoria Wiedemann) indicated that the LC50 of pyrethroids, as ppm in the wet weight, averaged 0.04 for deltamethrin pour-on, 0.25 for deltamethrin dip, 0.22 for alphacypermthrin pour-on, 0.10 for cyfluthrin pour-on, 0.23 for cypermethrin dip and 0.63 for flumethrin dip. Field bioassays involved artificial dung pats of 800 g, deployed in woodland and inspected after 24 h to record insects dead and alive. Beetles were most abundant in the wet season. Muscoid larvae were less seasonal. The LC50 of insecticides in the field confirmed laboratory indications. Adult Diptera (muscoids and Sgifidae) were not repelled or killed until the deltamethrin concentration reached 10 ppm. Pat dispersal by dung fauna and termites (Microtermes spp.) was halved by deltamethrin at 0.1-1 ppm. Scavenging of dead beetles by ants was greatest with small beetles (< 15 mm long) uncontaminated with insecticide. Dips and pour-ons of deltamethrin on cattle gave residues of about 0.01-0.1 ppm in dung produced in the fortnight after application. About 1.6% of the deltamethrin applied was transferred to dung. Deltamethrin and alphacypermethrin in dung showed no detectable degradation in 64 days. Contamination levels threaten populations of slow-breeding beetles.

Animals↗

Mark-recapture without estimating population sizes: a tool to evaluate termite baits.

The mark-recapture technique is currently being used by many termite researchers. Here its two uses are explained and disentangled. Use 1 is mapping the geographical location of the colony and use 2 is estimating the number of termites in the colony. Use 2 is controversial because the accuracy of the estimates has been widely questioned. In this article, it is explained that use 1 is not controversial and that use 1 (not use 2) is the criterion that is currently being used for evaluating the efficacy of termite baits.

Animals↗