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Background Termites primarily feed on lignocellulose or soil in association with specific gut microbes. The functioning of the termite gut microbiota is partly understood in a handful of wood-feeding pest species but remains largely unknown in other taxa. We intend to fill this gap and provide a global understanding of the functional evolution of termite gut microbiota. Results We sequenced the gut metagenomes of 145 samples representative of the termite diversity. We show that the prokaryotic fraction of the gut microbiota of all termites possesses similar genes for carbohydrate and nitrogen metabolisms, in proportions varying with termite phylogenetic position and diet. The presence of a conserved set of gut prokaryotic genes implies that essential nutritional functions were present in the ancestor of modern termites. Furthermore, the abundance of these genes largely correlated with the host phylogeny. Finally, we found that the adaptation to a diet of soil by some termite lineages was accompanied by a change in the stoichiometry of genes involved in important nutritional functions rather than by the acquisition of new genes and pathways. Conclusions Our results reveal that the composition and function of termite gut prokaryotic communities have been remarkably conserved since termites first appeared ~ 150 million years ago. Therefore, the “world’s smallest bioreactor” has been operating as a multipartite symbiosis composed of termites, archaea, bacteria, and cellulolytic flagellates since its inception.

Mid-Cretaceous amber from Myanmar, also known as “burmite”, has produced 1013 species of which 693 are Insects. The Mesozoic cockroach record is rich, with thousands of specimens that show diverse ecological adaptations. Cretaceous cockroaches in amber have been reported from Lebanon, Taimyr, New Jersey and France. Cockroaches from Myanmar amber are represented by at least 74 species, of which 14 are formally described. Spinaeblattina with S. myanmarensis gen. et sp. nov. and S. yixianensis comb. n. = Piniblattella yixianensis from the Yixian Formation is along with Cratovitisma the second genus of the Mesoblattinidae known from both amber and sedimentary deposits. Fossil ootheca are rare with 6 specimens reported so far and the present isolated one, Blattoothecichnus argenteus ichnogen. et ichnosp. nov., is the most ancient, portable, otherwise resembling that of Cryptocercus. Ancient crystals of organic salt, calcium oxalate, forming the ootheca protective layer have been preserved on its surface.

Lower termites, as well as their sister group, the subsocial wood-feeding cockroach Cryptocercus, rely on flagellated eukaryotic symbionts in the hindgut to cooperatively digest their wood diet. In Cryptocercus these flagellates undergo encystment cycles tightly coordinated with the molting cycle of their host, yet the resultant cysts play no demonstrated role in their transmission to neonates; the trophozoite stage of the flagellates is passed directly from parents to offspring via hindgut fluids (proctodeal trophallaxis). This pattern suggests that encystment is a vestige from a gregarious cockroach ancestor, when the flagellates had a functional, two-stage life cycle and the cysts were horizontally transmitted among hosts via coprophagy. The strong integration between flagellate encystment cycles and host developmental physiology in Cryptocercus indicates that the relationship of the flagellates with their proposed gregarious cockroach ancestor was not commensal, but parasitic, with flagellates likely obtaining benefits by taking advantage of host gut metabolites and ingested plant debris. When vertical transmission evolved the parasites were ‘captured,’ and their fitness became inescapably embedded in the fitness of their host. The vertical transmission of gut flagellates and the origin of host subsociality via proctodeal trophallaxis can be considered two sides of the same coin. From the host point of view proctodeal trophallaxis marks the origination of parental care by provisioning neonates with nourishment, metabolites and beneficial microbiota. From the flagellate point of view, proctodeal trophallaxis was a shift from horizontal to vertical transmission, pushing them from the parasitic to the mutualistic end of the symbiotic spectrum, arguably making this host behavioral change the most critical juncture in the evolutionary trajectory of the termite lineage.

The symbiotic gut flagellates of lower termites form host-specific consortia composed of Parabasalia and Oxymonadida. The analysis of their coevolution with termites is hampered by a lack of information, particularly on the flagellates colonizing the basal host lineages. To date, there are no reports on the presence of oxymonads in termites of the family Stolotermitidae. We discovered three novel, deep-branching lineages of oxymonads in a member of this family, the damp-wood termite Porotermes adamsoni. One tiny species (6-10 mu m), Termitimonas travisi, morphologically resembles members of the genus Monocercomonoides, but its SSU rRNA genes are highly dissimilar to recently published sequences of Polymastigidae from cockroaches and vertebrates. A second small species (9-13 mu m), Oxynympha loricata, has a slight phylogenetic affinity to members of the Saccinobaculidae, which are found exclusively in wood-feeding cockroaches of the genus Cryptocercus, the closest relatives of termites, but shows a combination of morphological features that is unprecedented in any oxymonad family. The third, very rare species is larger and possesses a contractile axostyle; it represents a phylogenetic sister group to the Oxymonadidae. These findings significantly advance our understanding of the diversity of oxymonads in termite guts and the evolutionary history of symbiotic digestion. (C) 2019 Elsevier GmbH. All rights reserved.

Cryptocercus is a genus of sub-social wood-feeding cockroaches that represents the sister group to the eusocial termites. We generated mitochondrial (12S+16S rRNA, COII), nuclear (28S rRNA) and Blattbacterium endosymbiont (16S+23S rRNA) sequence data for 8 new Chinese species, and combined these with previously available data to undertake the most extensive analysis of phylogenetic relationships within the genus to date. As expected, phylogenetic relationships among Blattabacterium strains were found to be congruent with those of their hosts. Three major clades were found to exist in Asian populations, one representing taxa from the Hengduan mountains in Southwestern China, a second including taxa from Russia, Korea, Northeastern China, and Yunnan in the Hengduan Mountains, and a third including taxa from the Qinling Mountains and Daba Mountains in Central China. A molecular dating analysis using 7 termite fossils to calibrate the molecular clock indicated that the divergence of American and Asian Cryptocercus occurred 55.09Ma (41.55-72.28Ma 95% CI), and that the radiations of American and Asian taxa occurred 28.48Ma (20.83-37.95Ma 95% CI) and 20.97Ma (15.78-27.21Ma 95% CI) respectively. Reconstruction of ancestral geographic distributions using S-DIVA suggested Cryptocercus was originally distributed across both continents, as opposed to ancestral migration of Cryptocercus from one continent to the other. The last common ancestor of Asian Cryptocercus was inferred to have existed in Central China. An examination of male chromosome numbers in Asian Cryptocercus showed that diploid numbers vary from 2n=15 to 2n=41, and indicates the presence of eight new species. Our study represents the most comprehensive phylogenetic and biogeographic study yet performed for this important group of cockroaches.

Split-brood field studies using the biparental, subsocial wood-feeding cockroach Cryptocercus punctulatus indicate that parental absence in families of three different age classes of nutritionally independent juveniles results in significantly slower growth when compared to sib groups allowed to remain with adults, even when nymphs are more than 2 years old when parents are removed. In one family that was divided when nymphs were 15 months old and followed for 4 years after treatment, 69 % of nymphs allowed to remain with parents reached maturity in the third year of the study, while those isolated from parents remained in the subadult stage at the 3-year mark. In the fourth year, measurements of wet weights and head capsule widths of the newly mature adults in the two treatments of this family were not significantly different. A parental vs. non-parental social environment, then, resulted in different rates of growth to reproductive maturity, but nymphs in both treatments ultimately reached similar adult sizes. The results indicate that parental effects can modify offspring ontogeny independently of the direct transfer of resources in the form of trophallactic food and symbionts. Because cockroaches in this genus are sister group to the developmentally plastic and juvenilized termites, further exploration of the role of the social environment on development in Cryptocercus is warranted. Termites are eusocial cockroaches that exhibit weak or delayed post-embryonic development; the majority of colony members are arrested in the small, soft-bodied, altricial morphotype displayed by early juvenile stages of their subsocial sister group, Cryptocercus. Developmental dynamics in this cockroach genus can therefore provide insight into conditions that may have favored the origin of eusociality in the lineage. Here, we demonstrate that the parental social environment in Cryptocercus punctulatus modifies offspring development independently of the direct transfer of hindgut fluids and does so for more than 2 years after hatch. A system in which parent-offspring interactions modify juvenile ontogeny, whatever the mechanism, contributes to the unique set of circumstances that ultimately resulted in the evolution of termite eusociality.

The complete mitogenome of the subsocial wood-eating cockroach Cryptocercus kyebangensis was sequenced and compared with that of another Asian cockroach, Cryptocercus relictus and the primitive termite Mastotermes darwiniensis. The complete mitogenome of C. kyebangensis is a closed circular molecule of 15,720 bp, containing13 protein-coding genes, two ribosomal RNA genes (12S rRNA and 16S rRNA), 22 transfer RNA genes, and one D-loop region. Nucleotide composition of the C. kyebangensis mitogenome shows an AT bias (74.4%), in particular, 46.0% A, 28.4% T, 9.7% G, and 15.9% C. GC skews are similar among the mitogenomes of C. kyebangensis, C. relictus, and M. darwiniensis, but AT skew is stronger in Cryptocercus mitogenomes than in the M. darwiniensis mitogenome. The whole mitogenome of C. kyebangensis showed 88.8% nucleotide similarity to that of C. relictus but only 74.9% similarity to that of M. darwiniensis. Phylogenetic analyses, based on amino acid sequences of the 13 mitochondrial proteins, showed tha...

Two new Chinese Cryptocercus (Blattodea: Cryptocercidae) species, C. arcuatus sp. nov. and C. convexus sp. nov., are described together with photographs. This description includes the external structure of the adults and structures of the female genitalia. A comparative study of female genitalia was based on the 5 Chinese species in the family Cryptocercidae. Significant differences in female genitalia of Cryptocercidae are summarized. A key to these 5 species and a checklist of the species of Cryptocercus worldwide are provided.

Members of the genus Trichonympha are among the most well-known, recognizable and widely distributed parabasalian symbionts of lower termites and the wood-eating cockroach species of the genus Cryptocercus. Nevertheless, the species diversity of this genus is largely unknown. Molecular data have shown that the superficial morphological similarities traditionally used to identify species are inadequate, and have challenged the view that the same species of the genus Trichonympha can occur in many different host species. Ambiguities in the literature, uncertainty in identification of both symbiont and host, and incomplete samplings are limiting our understanding of the systematics, ecology and evolution of this taxon. Here we describe four closely related novel species of the genus Trichonympha collected from South American and Australian lower termites: Trichonympha hueyi sp. nov. from Rugitermes laticollis, Trichonympha deweyi sp. nov. from Glyptotermes brevicornis, Trichonympha louiei sp. nov. from Calcaritermes temnocephalus and Trichonympha webbyae sp. nov. from Rugitermes bicolor. We provide molecular barcodes to identify both the symbionts and their hosts, and infer the phylogeny of the genus Trichonympha based on small subunit rRNA gene sequences. The analysis confirms the considerable divergence of symbionts of members of the genus Cryptocercus, and shows that the two clades of the genus Trichonympha harboured by termites reflect only in part the phylogeny of their hosts.

Complete mitochondrial genomes (mitogenomes) of two cockroach species, Periplaneta australasiae and Neostylopyga rhombifolia, 15,605 bp and 15,711 bp in length, respectively, were determined. As reported for other cockroach mitogenomes, the two mitogenomes possessed typical ancestral insect mitogenome gene composition and arrangement. Only several small intergenic spacers were found: one, which was common in all sequenced cockroach mitogenomes except for the genus Cryptocercus, was between tRNA-Ser (UCN) and ND1 and contained a 7bp highly conserved motif (WACTTAA). Three different types of short tandem repeats in the N. rhombifolia control region (CR) were observed. The homologous alignments of these tandem repeats with other six cockroach mitogenome CRs revealed a low similarity. Three conserved sequence blocks (CSB) were detected in both cockroach mitochondrial CRs. CSB1 was specific for blattinine mitogenomes and was highly conserved with 95% similarity, speculating that this block was a possible molecular synapomorphy for this subfamily. CSB3 located nearby downstream of CSB1 and has more variations within blattinine mitogenomes compared with CSB1. The CSB3 was capable of forming stable stem-loop structure with a small T-stretch in the loop portion. We assessed the influence of four datasets and two inference methods on topology within Orthopteroidea. All genes excluding the third codon positions of PCGs could generate more stable topology, and higher posterior probabilities than bootstrap values were presented at some branch nodes. The phylogenetic analysis with different datasets and analytical methods supported the monophyly of Dictyoptera and supported strongly the proposal that Isoptera should be classified as a family (Termitidae) of the Blattaria. Specifically, Shelfordella lateralis was inserted in the clade Periplaneta. Considering the K2P genetic distance, morphological characters, and the phylogenetic trees, we suggested that S. lateralis should be placed in the genus Periplaneta.

A comprehensive compendium on the taxonomy and biology of the 3106 living and fossil species of the worlds termites is presented, along with reviews of Isoptera morphology and evolution, identification keys, the history of termite systematics, and summary of the worlds 363 significant pest species. A complete bibliography is provided of nearly 5000 references covering virtually all aspects of termite taxonomy and biology through December 2011.The morphology of Isoptera is thoroughly reviewed and illustrated with original scanning electron micrographs and photomicrographs, covering the cuticular anatomy and those internal organs that are taxonomically and phylogenetically significant, including several new character systems. Terminology is presented for the systems of tibial spines and spurs so as to establish homologs. Keys are presented to the nine living families of termites, and the world subfamilies and genera of Archotermopsidae, Hodotermitidae, Kalotermitidae, Mastotermitidae, Rhinotermitidae, Serritermitidae, Stolotermitidae, and Stylotermitidae. A key to subfamilies of the Termitidae is included. A detailed morphological diagnosis for each family and subfamily is provided, along with images of exemplar species. The history of isopteran research in taxonomy, systematics, morphology, paleontology, and biology is reviewed from 1758 to the present, with emphasis on transformative workers such as Holmgren, Silvestri, Emerson, Roonwal, Noirot, and Sands. Evolution of the Isoptera is reviewed, including the diversity and natural history of genera and species in all Zoogeographic regions, major patterns in social biology, the phylogeny of Recent and fossil genera and families, and 135 million years of fossils preserved as compressions, mineralized replicas, and in amber. The definitive sister group to the Isoptera is the monogeneric family of wood roaches, Cryptocercidae (Cryptocercus), so the taxonomic ranks of the two groups are now Infraorder Isoptera and Infraorder Cryptocercoidea within Order Blattaria (roaches and termites).The compendium summarizes the taxonomic history, nomenclature, distribution, type locality, and repository, and all significant aspects of natural history and biology for each species of the world, exclusive of pest control and colony inquilines (termitophiles). The classification of Recent and fossil lower termites (all those exclusive of family Termitidae) used in the compendium is from Engel et al. (2009), which is based on morphology and largely congruent with molecular studies. Rhinotermitidae s.s. (exclusive of Stylotermitidae) may be paraphyletic with respect to Termitidae, although the six traditional subfamilies of the former are used here. A separate section summarizes the nomenclatural changes made in the compendium, including new synonymies, new combinations, status novus, lectotype selection etc. A detailed list is provided of museums and other institutional collections that house type specimens. An index is included. The Treatise is intended to provide an authoritative foundation for taxonomic work on the Isoptera, present and future.

The objective of this study is to elucidate the evolutionary rate of Cryptocercus SSU rDNAs based on the relative rate test (RRT) and comparative analysis of the primary sequences. In the RRTs, the evolutionary rate differences (dKs) of the Cryptocercus SSU rDNAs were not statistically significant (0.107

The wood-feeding cockroach Cryptocercus punctulatus Scudder (Blattodea: Cryptocercidae) is an important member of the dead wood (saproxylic) community in montane forests of the southeastern United States. However, its population biology remains poorly understood. Here, aspects of family group co-occurrence were characterized to provide basic information that can be extended by studies on the evolution and maintenance of sub-sociality. Broad sampling across the species’ range was coupled with molecular data (mitochondrial DNA (mtDNA) sequences). The primary questions were: (1) what proportion of rotting logs contain two or more different mtDNA haplotypes and how often can this be attributed to multiple families inhabiting the same log, (2) are multi-family logs spatially clustered, and (3) what levels of genetic differentiation among haplotypes exist within a log, and how genetically similar are matrilines of co-occurring family groups? Multi-family logs were identified on the premise that three different mtDNA haplotypes, or two different haplotypes among adult females, is inconsistent with a single family group founded by one male–female pair. Results showed that of the 88 rotting logs from which multiple adult C. punctulatus were sampled, 41 logs (47%) contained two or more mtDNA haplotypes, and at least 19 of these logs (22% overall) were inferred to be inhabited by multiple families. There was no strong evidence for spatial clustering of the latter class of logs. The frequency distribution of nucleotide differences between co-occurring haplotypes was strongly right-skewed, such that most haplotypes were only one or two mutations apart, but more substantial divergences (up to 18 mutations, or 1.6% uncorrected sequence divergence) do occasionally occur within logs. This work represents the first explicit investigation of family group co-occurrence in C. punctulatus, providing a valuable baseline for follow-up studies.

We identified the antifungal gene termicin in three species of Cryptocercus woodroaches. Cryptocercus represents the closest living cockroach lineage of termites, which suggests that the antifungal role of termicin evolved prior to the divergence of termites from other cockroaches. An analysis of Cryptocercus termicin and two β-1,3-glucanase genes (GNBP1 and GNBP2), which appear to work synergistically with termicin in termites, revealed evidence of selection in these proteins. We identified the signature of past selective sweeps within GNBP2 from Cryptocercus punctulatus and Cryptocercus wrighti. The signature of past selective sweeps was also found within termicin from Cryptocercus punctulatus and Cryptocercus darwini. Our analysis further suggests a phenotypically identical variant of GNBP2 was maintained within Cryptocercus punctulatus, Cryptocercus wrighti, and Cryptocercus darwini while synonymous sites diverged. Cryptocercus termicin and GNBP2 appear to have experienced similar selective pressure to that of their termite orthologues in Reticulitermes. This selective pressure may be a result of ubiquitous entomopathogenic fungal pathogens such as Metarhizium. This study further reveals the similarities between Cryptocercus woodroaches and termites.

We identified and characterized Gram-negative bacteria binding proteins (GNBPs) and their predicted antifungal activity in the woodroach Cryptocercus punctulatus. C. punctulatus is likely to share many characteristics with the subsocial ancestor of the Isoptera, including allogrooming, which facilitated the evolution of termite sociality. Based on a phylogenetic analysis, an ancestral GNBP with an intact β-1,3-glucanase active site appears to have duplicated in a common ancestor of subsocial Cryptocercus woodroaches and termites. In termites, the secreted β-1,3-glucanase activity of GNBPs provides important prophylactic protection from fungal pathogens such as Metarhizium anisopliae, which can evade the immune system after entering the insect. Here, we identify β-1,3-glucanase activity on the cuticular surface of C. punctulatus that originates from the salivary gland and is likely spread by allogrooming. Cuticular washes have antifungal activity against M. anisopliae conidia that is suppressed by an inhibitor (GDL) of termite GNBP β-1,3-glucanase activity. C. punctulatus nymphs that are treated with GDL and subsequently exposed to M. anisopliae conidia show significantly greater mortality than the untreated nymphs exposed to conidia. A molecular evolutionary analysis of GNBPs in two species of Parcoblatta, Periplaneta americana, C. punctulatus and representative termites indicates that selection-directed change in a glycosylphosphatidylinositol (GPI) anchor region. Modification of the GPI anchor region may have been instrumental in the evolution of an antifungal defense strategy that depends on the external secretion of GNBPs from the salivary gland and their dissemination by grooming. This strategy may have helped compensate for the vulnerability of a subsocial woodroach-like ancestor to fungal disease that results from prolonged development with a thin cuticle and facilitated the transition to termite eusociality.

Anaerobic cellulolytic flagellate protists of the hindguts of lower termites and the wood-feeding cockroach Cryptocercus are essential to their host's ability to digest lignocellulose. Many have bacteria associated with their surfaces and within cytoplasmic vesicles—likely important symbioses as suggested by molecular and other data. Some of the most striking examples of these symbioses are in the parabasalid family Hoplonymphidae, but little or no data exist on the structural aspects of their symbioses, their relationships with bacteria through different life-cycle stages, or their diversity and phylogenetic relationships in Cryptocercus. We investigated these areas in the hoplonymphid genera Barbulanympha and Urinympha from Cryptocercus punctulatus using light and electron microscopy, and analysis of small subunit rRNA. Microscopy reveals variation in density of bacterial surface symbionts related to life-cycle stage, a glyococalyx possibly important in bacterial adhesion and/or metabolite exchange, and putative viruses associated with bacterial surface symbionts. Patterning of surface bacteria suggests protists emerging from the resistant (dormant) stage are colonized by a small population of bacterial cells, which then divide to cover their surface. Additionally, cytoplasmic protrusions from the protist are covered by bacteria. Phylogenetic analysis rejects the monophyly of Hoplonymphidae, suggesting multiple origins or losses of these bacterial symbioses.

Wood-feeding cockroaches in the genus Crytocercus live in biparental family groups in the interior of rotten logs and are considered socially monogamous. They typically produce a single brood of altricial offspring, with parental care then lasting until the death of adults 2 or 3 years later. Field and laboratory evidence suggests that copulation occurs repeatedly in C. punctulatus, with sexual activity occurring during non-fertile periods. We carried out a long-term field study to explore the significance of repeated copulation in C. punctulatus. The presence of a male positively affected the onset of female reproduction, but neither the male’s physical presence nor his ability to mate influenced the number or early development of nymphs. Sperm are viable in the spermathecae for at least 3 years. We conclude that it is unlikely that male-derived material benefits have driven the evolution of repeated copulation in C. punctulatus. Accumulating evidence suggests that prior to oviposition, repeated copulation may be rooted in sperm competition, but after the eggs are laid it may secure the continued paternal contribution of the male social partner.

A completely preserved cockroach Sociala perlucida gen. et sp. nov. (Blattida: Socialidae fam. nov.) is described from the Mesozoic (Albian Early Cretaceous) amber of Archingeay in France. It is categorized within the new family Socialidae, originating from the Mesozoic cockroach family Liberiblattinidae, and representing the sister group to the most basal known eutermite families. Numerous direct and indirect morphological evidence (such as a unique narrow body and pronotum with reduced coloration, legs without carination and with numerous sensillar pitts (forelegs) as adaptations to life in nests analogical to the living termitophillous species; enlarged head, long palps for communication, a general venation modified in a direction towards termites (subcosta and radial vein [R] approximated, R branches approximated, simplified, cross-veins reduced), and possibly also the development of the breaking forewing sutura for detaching wings after marriage flight), suggest its eusocial mode of life. Thus, the first eusociality originated within cockroaches, prior to giving birth to termites; termites and Cryptocercus are not directly related. The group of eusocial cockroaches makes termite morphotaxon polyphyletic, but also in this case, the erection of a new order for mastotermites (including Cratomastotermitidae) and/or the inclusion of eusocial, morphological cockroaches within termites appears counterproductive, thus an example of such a general exception to the taxonomical procedure is provided.

Parabasalia are a large, diverse clade of anaerobic flagellates, many of which inhabit the guts of wood-feeding insects. Because most are uncultivable, molecular data representing the true diversity of Parabasalia only became possible with the application of single-cell techniques, but in the last decade molecular data have accumulated rapidly. Within the Trichonymphida, the most diverse lineage of hypermastigote parabasalids, molecular data are now available from five of the six families, however, one family, the Spirotrichosomidae, has not been sampled at the molecular level, and is very little studied with electron microscopy. Here we examine a representative of Spirotrichosomidae--Leptospironympha of the wood-feeding cockroach Cryptocercus punctulatus--with scanning and transmission electron microscopy, and analyze its small subunit rRNA gene to infer its phylogenetic position. Phylogenetic analyses place Leptospironympha as sister to a clade comprising Eucomonymphidae and Teranymphidae with moderate support. Examination with scanning and transmission electron microscopy reveals new classes of previously undetected symbiotic surface bacteria, a glycocalyx, granular particles on flagella, and putative phagocytosed bacteria. The range of flagellar patterns in Spirotrichosomidae is quite wide, and the possibility that some members may be more closely related to Eucomonymphidae or Teranymphidae is addressed.

Subsocial wood feeding cockroaches in the genus Cryptocercus, the sister group of termites, retain their symbiotic gut flagellates during the host molting cycle, but in lower termites, closely related flagellates die prior to host ecdysis. Although the prevalent view is that termite flagellates die because of conditions of starvation and desiccation in the gut during the host molting cycle, the work of L.R. Cleveland in the 1930s through the 1960s provides a strong alternate hypothesis: it was the changed hormonal environment associated with the origin of eusociality and its concomitant shift in termite developmental ontogeny that instigates the death of the flagellates in termites. Although the research on termite gut microbial communities has exploded since the advent of modern molecular techniques, the role of the host hormonal environment on the life cycle of its gut flagellates has been neglected. Here Cleveland’s studies are revisited to provide a basis for re-examination of the problem, and the results framed in the context of two alternate hypotheses: the flagellate symbionts are victims of the change in host social status, or the flagellates have become incorporated into the life cycle of the eusocial termite colony. Recent work on parasitic protists suggests clear paths for exploring these hypotheses and for resolving long standing issues regarding sexual-encystment cycles in flagellates of the Cryptocercus-termite lineage using molecular methodologies, bringing the problem into the modern era.

Two Early Cretaceous Burmese amber cockroaches contained protists related to mutualistic flagellates occurring in extant Cryptocercus cockroaches and lower termites. The fossil protists are described as Devescovites proteus Poinar n. gen., n. sp. (Parabasalia: Trichomonadida: Devescovinidae), Paleotrichomones burmanicus Poinar n. gen., n. sp. (Parabasalia: Trichomonida), Burmanymphus cretacea Poinar n. gen., n. sp. (Hypermastigia: Trichonymphida: Burmanymphidae n. fam.) and Oxymonas gigantea Poinar, n. sp. Additional putative protists are also illustrated. Evolutionary implications of this discovery are discussed.

Species in the wood-feeding genus Salganea within the cockroach subfamily Panesthiinae (Blaberidae) typically live in biparental families; their first instars suffer high mortality when removed from adults, and in at least one species, adults are known to feed neonates on oral liquids. In the closely related gregarious wood-feeding genus Panesthia , no parental interaction with offspring is known. We compared the external morphology of first instars of these two genera and found that eye development and cuticular pigmentation at hatching are correlated with social structure. First instars of Panesthia have a dark cuticle and well-developed eyes. First instars of Salganea have a pale, transparent cuticle, and eyes significantly smaller than those of Panesthia relative to adult eye size. The body size of the first-instar of Salganea esakii is significantly smaller than that of Panesthia angustipennis spadica , relative to adult body size (24.0 and 27.4% of adult head-capsule width, respectively), but a more general survey suggests that, overall, neonate size may be similar in the two genera. We compared the first instars of these two taxa of Panesthiinae to those in the biparental, wood-feeding cockroach Cryptocercus (Cryptocercidae) and discuss how decreased investment in both integumentary and ocular development in subsocial cockroaches parallels that seen in altricial vertebrates.

Cryptocercus cockroaches and lower termites harbour obligate, diverse and unique symbiotic cellulolytic flagellates in their hindgut that are considered critical in the development of social behaviour in their hosts. However, there has been controversy concerning the origin of these symbiotic flagellates. Here, molecular sequences encoding small subunit rRNA and glyceraldehyde-3-phosphate dehydrogenase were identified in the symbiotic flagellates of the order Trichonymphida (phylum Parabasalia) in the gut of Cryptocercus punctulatus and compared phylogenetically to the corresponding species in termites. In each of the monophyletic lineages that represent family-level groups in Trichonymphida, the symbionts of Cryptocercus were robustly sister to those of termites. Together with the recent evidence for the sister-group relationship of the host insects, this first comprehensive study comparing symbiont molecular phylogeny strongly suggests that a set of symbiotic flagellates representative of extant diversity was already established in an ancestor common to Cryptocercus and termites, was vertically transmitted to their offspring, and subsequently became diversified to distinct levels, depending on both the host and the symbiont lineages.

Recent phylogenetic studies indicating that termites are eusocial cockroaches closely related to the genus Cryptocercus have generated fresh interest in wood-feeding Blattaria. Here we summarize the social biology of the wood-feeding genus Salganea (Blaberidae: Panesthiinae). As in Cryptocercus, Salganea exhibits long term, biparental care that includes the defense and feeding of young nymphs. Unlike Cryptocercus, however, Salganea is iteroparous: most studied species reproduce periodically over their lifetime. These divergent reproductive strategies are likely related to parental costs associated with their differing reproductive modes: Cryptocercus is oviparous, while Salganea is ovoviviparous. The pattern of parental investment associated with ovoviviparous reproduction may be one reason why Salganea and other ovoviviparous wood-feeding cockroach lineages did not evolve eusociality.

Nitrogen fixation by gut microorganisms is one of the crucial aspects of symbiosis in wood-feeding termites since these termites thrive on a nitrogen-poor diet. In order to understand the evolution of this symbiosis, we analysed the nitrogenase structural gene nifH in the gut microbial communities. In conjunction with the published sequences, we compared approximately 320 putatively functional NifH protein sequences obtained from a total of 19 termite samples that represent all the major branches of their currently proposed phylogeny, and from one species of the cockroach Cryptocercus that shares a common ancestor with termites. Using multivariate techniques for clustering and ordination, a phylogeny of NifH protein sequences was created and plotted variously with host termite families, genera, and species. Close concordance was observed between NifH communities and the host termites at genus level, but family level relationships were not always congruent with accepted termite clade structure. Host groups examined included basal families (Mastotermitidae, Termopsidae, Kalotermitidae, as well as Cryptocercus), the most derived lower termite family Rhinotermitidae, and subfamilies representing the advanced and highly diverse apical family Termitidae (Macrotermitinae, Termitinae, and Nasutitermitinae). This selection encompassed the major nesting and feeding styles recognized in termites, and it was evident that NifH phylogenetic divergence, as well as the occurrence of alternative nitrogenase-type NifH, was to some extent dependent on host lifestyle as well as phylogenetic position.

Termites are instantly recognizable mound-builders and house-eaters: their complex social lifestyles have made them incredibly successful throughout the tropics. Although known as ‘white ants’, they are not ants and their relationships with other insects remain unclear. Our molecular phylogenetic analyses, the most comprehensive yet attempted, show that termites are social cockroaches, no longer meriting being classified as a separate order (Isoptera) from the cockroaches (Blattodea). Instead, we propose that they should be treated as a family (Termitidae) of cockroaches. It is surprising to find that a group of wood-feeding cockroaches has evolved full sociality, as other ecologically dominant fully social insects (e.g. ants, social bees and social wasps) have evolved from solitary predatory wasps.

The hindguts of lower termites and Cryptocercus cockroaches are home to a distinct community of archaea, bacteria, and protists (primarily parabasalids and some oxymonads). Within a host species, the composition of these hindgut communities appears relatively stable, but the evolutionary and ecological factors structuring community composition and stability are poorly understood, as are differential impacts of these factors on protists, bacteria, and archaea. We analyzed the microbial composition of parabasalids and bacteria in the hindguts of Cryptocercus punctulatus and 23 species spanning 4 families of lower termites by pyrosequencing variable regions of the small-subunit rRNA gene. Especially for the parabasalids, these data revealed undiscovered taxa and provided a phylogenetic basis for a more accurate understanding of diversity, diversification, and community composition. The composition of the parabasalid communities was found to be strongly structured by the phylogeny of their hosts, indicating the importance of historical effects, although exceptions were also identified. Particularly, spirotrichonymphids and trichonymphids likely were transferred between host lineages. In contrast, host phylogeny was not sufficient to explain the majority of bacterial community composition, but the compositions of the Bacteroidetes , Elusimicrobia , Tenericutes , Spirochaetes , and Synergistes were structured by host phylogeny perhaps due to their symbiotic associations with protists. All together, historical effects probably resulting from vertical inheritance have had a prominent role in structuring the hindgut communities, especially of the parabasalids, but dispersal and environmental acquisition have played a larger role in community composition than previously expected.

Members of the genus Cryptocercus are wood-feeding cockroaches that live in the temperate forests. Nine species are recognized in the genus worldwide: two in eastern Eurasia, two in China, and five in the United States. Within the United States, one species occurs in the Pacific Northwest and four occur in the Appalachian Mountains. Previous studies have revealed the presence of potential zones of overlap in distribution among the Appalachian species, raising the possibility of hybridization among them. Differences in mitochondrial DNA have previously been identified for the Appalachian species. However, to identify hybrid individuals one or more species-specific, codominant nuclear markers are required. Therefore, our objective was to undertake allozyme analysis of enzymatic loci to identify fixed, species-specific alleles for the four Appalachian species. We assayed a mean of 42 individuals each from 16 sites for allozyme variation for the four species. At 6 of the 33 loci examined, fixed alternate alleles were identified; a combination of 2 loci enabled the identification of all four species. To identify hybrids in the field, we examined 42 individuals each from 13 sites in which two or more of the above species occur in close proximity for presence of heterozygous individuals at one or more of the six fixed loci. No heterozygous individuals were found suggesting the lack of hybridization among the Appalachian species.

Termites of most isopteran families and wood-feeding cockroaches of the genus Cryptocercus usually harbor more than one symbiotic flagellate species in their hindgut. To evaluate the similarity of their symbiont faunae, data on symbiont composition at a generic level were examined by cluster analysis and type III quantification method. In both analyses, the symbiont composition recorded from host insects belonging to the same families or monophyletic family groups tended to be similar. This tendency was particularly remarkable in the clade Kalotermitidae and the clade Rhinotermitidae plus Serritermitidae. Two basal host groups, the Cryptocercidae and the Mastotermitidae, exhibited very different symbiont compositions. These findings suggested that the symbiont faunae mainly reflect the host's phylogenetic relationships. Within the Rhinotermitid hosts, the genus Reticulitermes showed a unique symbiont fauna although it is not a basal taxon in the Rhinotermitidae. Horizontal transfers of symbiotic protists might explain such anomalistic fauna.

The distributional pattern of the four known karyotypes (male 2n = 37, 39, 43, 45) of the Cryptocercus punctulatus Scudder species complex is reported, based on 71 sites in the Southern Appalachian Mountains with an emphasis on western North Carolina. Populations with different karyotypes are geographically structured in a mosaic, with at least one karyotype occurring in two disjunct regions. Abrupt geographic transitions between karyotypes suggest a parapatric distribution. We found no overlap in the distribution of the different karyotypes, as recently suggested. Although the boundary zones between karyotypes do not appear to coincide with physical or ecological barriers to dispersal, several transitions between karyotypes occur on or near the highest mountains in the southern Appalachians. We suggest that the different karyotypes arose by vicariance, with current boundaries formed by secondary contact when populations isolated in glacial refugia subsequently spread into high-mountain habitats. Because of their dependence on mature mesic forests, populations of the cockroach likely advance up and down mountainsides in cycles of advances and retreats dictated by climatic oscillations that raise and lower the timberline. We discuss the taxonomic status of the different karyotype groups in the C. punctulatus complex, and conclude that more exacting evidence is required to establish if species-level status is warranted. The conclusions of certain earlier studies are weak because, among other things, karyology was not examined in the sampled specimens, including those designated as types.

Modern termite phylogenetics is critically reviewed, with an emphasis on tree topologies as phylogenetic hypotheses. Studies have especially concentrated on (1) the position of Isoptera among the Dictyoptera and (2) the family group relationships within the Isoptera. The first of these problems is still controversial; although the weight of evidence now suggests that termites are nested within the cockroaches, thus making "Blattaria" as presently constituted paraphyletic. The exact position of termites within the cockroaches is uncertain, although Cryptocercus is the most plausible sister group.¶Family groups relationships are rather better resolved. Mastotermitidae is now generally accepted to be the most basal termite group. Termopsidae, Hodotermitidae and Kalotermitidae are all basal to (Termitidae + Serritermitidae + Rhinotermitidae), although their relative positions within that part of the tree are disputed. Most recent studies support a sister group relationship for Serritermitidae and (Termitidae + Rhinotermitidae). However, no study has yet unambiguously found the Rhinotermitidae monophyletic. The Termitidae are well established as monophyletic and as the most apical termite family. However, within the Termitidae the monophyly of none of the subfamilies is well established, making subfamily level analyses unreliable.¶A number of problem areas are identified: (1) poor taxon sampling is a universal problem, (2) higher taxonomic groupings are often assumed to be monophyletic a priori without adequate support, (3) datasets are collected from different taxa and character systems without consideration of the overall international effort.

Cryptocercus are subsocial, xylophagous cockroaches that live in temperate forests. Like other cockroaches, Cryptocercus harbour endosymbiotic bacteria in their fat bodies. Two species of Cryptocercus occur in the palaearctic, one each in eastern Russia and south-central China. In the USA, there are five species: one in the north-west and four in the south-east. Little is known about the relationship between the Eurasian and North American Cryptocercus or the causes of the disjunct distribution. Here, a molecular phylogeny for six out of the seven Cryptocercus species and their endosymbionts is inferred in an attempt to understand the evolution and biogeography of the genus. Our analysis showed that the North American Cryptocercus are monophyletic, suggesting that a single colonization event was followed by vicariance. There was complete concordance between the host and endosymbiont phylogenetic trees. Divergence estimates based on endosymbiont DNA sequences suggested that the palaearctic and nearctic Cryptocercus diverged 70-115 million years (Myr) ago and the eastern- and western-USA species diverged 53-88 Myr ago. These divergence estimates were correlated with biogeographical events, and a hypothesis is presented to explain the current distribution of Cryptocercus. Our findings suggest that Cryptocercus has had a long evolutionary history, dating back to the Jurassic.

Summary Phylogenetic relationships among five cockroach families (Cryptocercidae, Polyphagidae, Blattidae, Blattellidae and Blaberidae) using seventeen species, were estimated based on the DNA sequences of the mitochondrial cytochrome oxidase subunit II (COII) gene. A cladogram inferred using the neighbour-joining method indicated that Polyphagidae and Cryptocercidae are closely related to each other, and that these two groups are a sister group to the remaining cockroach families. The monophyly of this clade, however, was not strongly supported in terms of bootstrap percentages. Blaberidae and Blattellidae were shown to be sister groups as previously proposed, with Blattidae as a sister group to that clade. Non-weighted and weighted parsimony analyses were also performed following analyses of nucleotide substitution patterns that indicated saturation of the COII gene among these taxa had occurred. These parsimonious cladograms suggested that Polyphagidae was the basal family, and Polyphaginae, including Cryptocercus as proposed by Grandcolas 1994a), was not monophyletic. The inferred relationships among cockroach families (Polyphagidae, Cryptocercidae + (Blattidae + (Blattellidae + Blaberidae))) is partly in agreement with some previously published analyses. Additionally, based on molecular data, Asian and American Cryptocercus are suggested to have diverged from one another before the Oligocene (∼20 mya).

Recent phylogenetic studies showed that the genus Cryptocercus belongs to the subfamily Polyphaginae. This result is well supported by the total congruence between these analyses which were independently carried out on morphology and on neuropeptides from corpora cardiaca. Other recent studies contradict each, other and are based on incorrectly defined characters or inappropriate methodologies. Preconceived ideas concerning Cryptocercus (primitiveness, relict distribution) definitely need to be confronted with facts. Facts indicate that Cryptocercus has a surprisingly high dispersal ability and that its present preferenda and distribution suggest a late Tertiary origin. Speculations about unwarrantedly clock-like evolution and dubious fossil records should not be preferred to both congruent phylogenetic framework and facts.

The evolution of pre-social behaviour in two clades of cockroaches was studied in reference to phylogeny. In each clade, both solitary and pre-social behaviours exist. These behaviours are parsimoniously mapped on phylogenies obtained from previous morpho-anatomical studies. Solitariness appears to have evolved several times from pre-social ancestors. These reversals in pre-social behaviour stress once more the usefulness of phylogeny for revealing unexpected patterns, similar to the results obtained in studies of behavioural evolution in Hymenoptera. A simple model involving lack of adaptive radiation in solitaryderived clades and small size of selected ingroups accounts for the observed pattern of common evolutionary reversals.

Seventy-two morpho-anatomical characters were examined in 221 genera belonging to the families Blattidae, Polyphagidae, Blattellidae, and Blaberidae. They were cladistically analyzed and polarized using two mantids and two termites. As no autapomorphies of the family Blattellidae were found, the constituent subfamilies were used as terminal taxa together with other families. Three trees were found (CI = 0.81 and RI = 0.88, without autapomorphies) that differed only by the position of Nyctiborinae relative to Blattellinae and Ectobiinae. The strict consensus tree was [Blattidae [Polyphagidae [Anaplectinae [[Pseudophyllodromiidae, Blaberidae] [Nyctiborinae, Blattellinae, Ectobiinae]]]]]. The main discrepancies with McKittrick's tree were the monophyly of Polyphagidae (instead of paraphyly) and that the Blaberidae is the sister-group of Pseudophyllodromiinae (instead of the sister-group of Blattellinae, Ectobiinae, and Nyctiborinae). These results made it necessary to elevate the Anaplectinae and Pseudophyllodromiinae to familial status, and to give a new sense to the family Blattellidae, which includes only the subfamilies Blattellinae, Ectobiinae, and Nyctiborinae. This phylogeny was used to test current evolutionary hypotheses concerning sociality and reproductive behaviour; many traits were assumed to be ancestral to all cockroaches (protozoan symbionts and familial life habits) or preadaptive (progressing from advanced oviparity in Blattellidae to ovoviviparity in Blaberidae) that must actually be considered homoplasic.

We have designed and utilized degenerate primers in the phylogenetic analysis of [FeFe] hydrogenase gene diversity in the gut ecosystems of roaches and lower termites. H(2) is an important free intermediate in the breakdown of wood by termite gut microbial communities, reaching concentrations in some species exceeding those measured for any other biological system. The primers designed target with specificity the largest group of enzymatic H domain proteins previously identified in a termite gut metagenome. "Family 3" hydrogenase sequences were amplified from the guts of lower termites, Incisitermes minor, Zootermopsis nevadensis, and Reticulitermes hesperus, and two roaches, Cryptocercus punctulatus and Periplaneta americana. Subsequent analyses revealed that all termite and Cryptocercus sequences were phylogenetically distinct from non-termite-associated hydrogenases available from public databases. The abundance of unique sequence operational taxonomic units (as many as 21 from each species) underscores the previously demonstrated physiological importance of H(2) to the gut ecosystems of these wood-feeding insects. The diversity of sequences observed might be reflective of multiple niches that the enzymes have been evolved to accommodate. Sequences cloned from Cryptocercus and the lower termite samples, all of which are wood feeding insects, clustered closely with one another in phylogenetic analyses to the exclusion of alleles from P. americana, an omnivorous cockroach, also cloned during this study. We present primers targeting a family of termite gut [FeFe] hydrogenases and provide results that are consistent with a pivotal role for hydrogen in the termite gut ecosystem and point toward unique evolutionary adaptations to the gut ecosystem.

Subsocial, xylophagous cockroaches of the genus Cryptocercus exhibit a disjunct distribution, with representatives in mature montane forests of North America, China, Korea and the Russian Far East. All described species are wingless and dependent on rotting wood for food and shelter at all stages of their life cycle; consequently, their distribution is tied to that of forests and strongly influenced by palaeogeographical events. Asian and American lineages form distinct monophyletic groups, comprised of populations with complex geographic substructuring. We review the phylogeny and distribution of Cryptocercus, and discuss splitting events inferred from molecular data.

The genus Therea Billberg, 1820 is redefined as a monophyletic taxon belonging to the group Ergaula + Eucorydia + Therea + Cryptocercus in the subfamily Polyphaginae Kirby, 1904. Three new species, T. regularis, T. defranceschii, and T. hyperguttata, are described as well as T. petiveriana (Linnaeus, 1758) and T. nuptialis (Gerstaecker, 1861). The known distribution of the genus in India extends from dry deciduous forests to more humid formations (between latitudes 19 and 10° N). The species live in areas with mean annual rainfall ranging from 500 to 3800 mm.

Cockroaches show the entire range of reproductive modes: oviparous, ovoviviparous, viviparous, and intermediate stages. Postparturition parental care is likewise diverse, ranging from species in which females remain with neonates for a few hours, to biparental care that lasts several years and includes feeding the offspring on bodily fluids in a nest. Both ovoviviparity and parental care arose a number of times in the taxon. Evolution of reproductive mode seems most influenced by predators, parasites and cannibalism. Ovoviviparity, aggregation behavior of young nymphs, and diet are suggested as factors influential in the evolution of postparturition parental care. Females regulate parental investment via absorption of oocytes, abortion, cannibalism and brood reduction. The developmental status of cockroaches at hatching ranges along an altricial–precocial spectrum and is correlated with the presence and type of parental care. In several subsocial species neonates are blind, poorly sclerotized, and dependent for food, while in the sole viviparous cockroach nymphs hatch in an advanced state of development and require fewer molts to adulthood than any known cockroach. Association with microorganisms in both the digestive system and the fat body is suggested as one factor influential in the repro – ductive versatility of cockroaches. In particular, the endosymbiont flavobacteria which mediate the storage and recycling of nitrogenous waste products may allow for the variety of modes of postovulation provisioning of offspring.

SUMMARY Basal termites possess two developmental features that eusocial Hymenoptera lack: the majority of colony members are juveniles whose somatic and reproductive development is temporarily or permanently suspended, and individual development is characterized by extreme phenotypic plasticity. An examination of the literature indicates that the basis for these unique ontogenetic characters is not the prolongation of a pronymphal stage into postembryonic development, as recently suggested. Like other hemimetabolous insects, termites have three embryonic cuticles, and the pronymphal (EC3) cuticle is shed during or shortly after hatch. Nonetheless, a different developmental landmark, dorsal closure, occurs later during embryogenesis in termites than it does in their cockroach relatives, clearly indicating ontogenetic repatterning from an ancestral state. An alternate hypothesis for the origin of isopteran phenotypic plasticity becomes apparent if we remain focused on the phylogenetic and social context of termite evolution. Altricial development occurs in both vertebrate and invertebrate taxa, evolves in response to the parental environment, and is displayed by two distantly related, biparental, wood-feeding cockroaches, including Cryptocercus, the sister-group to termites. It is therefore likely the condition was present in subsocial termite ancestors, and played a complex, multidimensional role in the transition to eusociality. Most relevant to current arguments is that a shift in responsibility for the care of altricial dependents, from parents to the first nutritionally independent nymphs in the family (alloparents), resulted in the developmental stasis of alloparents at a relatively young age. Because early instar cockroaches are not metamorphically competent, these young alloparents would have provided a novel developmental template on which selection could act.

Termites are key decomposer insects in numerous ecosystems in the tropics and beyond, and their unique social systems provide a major counterpoint to those of hymenopteran social insects. Our knowledge of the phylogenetics and systematics of the group have traditionally lagged behind those of other important insect groups, but significant progress has now been made. Here we review recent phylogenetic studies of relationships both among termites, and between termites, cockroaches and mantids. We also discuss studies of co-cladogenesis between termites and two groups of symbionts: cellulolytic hindgut flagellates, and Blattabacterium. A consensus has emerged that the sister-group of termites is the wood-feeding cockroach genus Cryptocercus, and that the digestion of wood by the common ancestor of these two groups was aided by cellulolytic hindgut flagellates. The basal phylogenetic position of Mastotermes darwiniensis among termites has been confirmed, however agreement on the phylogenetic positions of members of the Kalotermitidae, Termopsidae and Hodotermitidae has yet to be reached. Relationships between and within the Rhinotermitidae and Serritermitidae also remain to be settled. Key lineages of the major family Termitidae, however, are now fairly well established.

Altricial development, the production of vulnerable, dependent neonates, is regularly associated with high levels of parental care in both vertebrates and invertebrates and has evolved in at least two phylogenetically distant, biparental wood-feeding cockroach genera: Cryptocercus and Salganea. Here I make the case that altricial development was the first deviation from standard hemimetabolous development in the biparental termite ancestor, played a fundamental role in the origin of the termite lineage, and can account for many of the novel traits and functions that constitute eusociality in extant Isoptera. Altricial neonates of subsocial wood feeding cockroaches are small, fragile, incapable of feeding themselves, and costly to rear. However, the type of care these juveniles require does not necessarily have to originate from a parent; consequently other members of the social group have the opportunity to undertake brood care obligations. Altricial development therefore allows for the transition to alloparental care, and has a great deal of explanatory power when the evolution of termite eusociality is framed as a hierarchical two-stage process. Stage 1 alloparental care both increased reproductive output of adults and prolonged the development of young helpers. Combined, this changed the nature of the family: it was composed of a greater number of small, vulnerable juveniles. At the same time however, parents were increasingly specializing in reproduction at the expense of their defensive parental duties. Stage 1 alloparental care, then, necessitated Stage 2, the development of a soldier caste, but it also allowed it. Soldiers are a nutritionally dependent caste, so alloparental feeding would have to be part of the behavioral repertoire before soldiers with specialized defensive mandibles could appear. As eusociality progressed, altricial development formed the operational basis for the prolongation of juvenile morphology, physiology and behavior into later ontogeny, accounting for many aspects of termite life history neglected by current hypotheses. Altricial development reinforces the importance of biparental care and hemimetabolous development in the termite ancestor and is a clear indication that the social environment can significantly influence developmental ontogeny in the lineage. The evolution of termite eusociality is a prime example of how evolution uses existing components in the origin of new systems.

Relationships among six key dictyopteran taxa (Mantodea; Blat- todea (excluding Cryptocercidae); Cryptocercidae; Mastotermes darwiniensis, Termopsidae and Kalotermitidae (Isoptera)) are analysed based on seventy morphological, developmental and behavioural characters. The fossil record and the 'living fossil' genera Cryptocercus, Mastotermes and Archotermopsisare discussed in detail. Exact analysis of the character state matrix by implicit enu- meration (Hennig86) resulted in one cladogram, strongly supporting Blattodea + Cryptocercidae as a sister group to Mantodea, with the Isoptera as a sister group to that complex. Arrangements within the termites are equivocal, with Termopsidae and Mastotermes darwiniensis possible as the relatively most pri- mitive element of Isoptera.

Trichonympha is one of the most complex and visually striking of the hypermastigote parabasalids-a group of anaerobic flagellates found exclusively in hindguts of lower termites and the wood-feeding cockroach Cryptocercus-but it is one of only two genera common to both groups of insects. We investigated Trichonympha of Cryptocercus using light and electron microscopy (scanning and transmission), as well as molecular phylogeny, to gain a better understanding of its morphology, diversity, and evolution. Microscopy reveals numerous new features, such as previously undetected bacterial surface symbionts, adhesion of post-rostral flagella, and a distinctive frilled operculum. We also sequenced small subunit rRNA gene from manually isolated species, and carried out an environmental polymerase chain reaction (PCR) survey of Trichonympha diversity, all of which strongly supports monophyly of Trichonympha from Cryptocercus to the exclusion of those sampled from termites. Bayesian and distance methods support a relationship between Trichonympha species from termites and Cryptocercus, although likelihood analysis allies the latter with Eucomonymphidae. A monophyletic Trichonympha is of great interest because recent evidence supports a sister relationship between Cryptocercus and termites, suggesting Trichonympha predates the Cryptocercus-termite divergence. The monophyly of symbiotic bacteria of Trichonympha raises the intriguing possibility of three-way co-speciation among bacteria, Trichonympha, and insect hosts.

Symbiotic flagellates play a major role in the digestion of lignocellulose in the hindgut of lower termites. Many termite gut flagellates harbour a distinct lineage of bacterial endosymbionts, so-called Endomicrobia, which belong to the candidate phylum Termite Group 1. Using an rRNA-based approach, we investigated the phylogeny of Trichonympha, the predominant flagellates in a wide range of termite species, and of their Endomicrobia symbionts. We found that Trichonympha species constitute three well-supported clusters in the Parabasalia tree. Endomicrobia were detected only in the apical lineage (Cluster I), which comprises flagellates present in the termite families Termopsidae and Rhinotermitidae, but apparently absent in the basal lineages (Clusters II and III) consisting of flagellates from other termite families and from the wood-feeding cockroach, Cryptocercus punctulatus. The endosymbionts of Cluster I form a monophyletic group distinct from many other lineages of Endomicrobia and seem to have cospeciated with their flagellate host. The distribution pattern of the symbiotic pairs among different termite species indicates that cospeciation of flagellates and endosymbionts is not simply the result of a spatial separation of the flagellate lineages in different termite species, but that Endomicrobia are inherited among Trichonympha species by vertical transmission. We suggest extending the previously proposed candidatus name 'Endomicrobium trichonymphae' to all Endomicrobia symbionts of Trichonympha species, and estimate that the acquisition by an ancestor of Trichonympha Cluster I must have occurred about 40-70 million years ago, long after the flagellates entered the termites.

The evolution of genome size has been discussed in relation to the evolution of various biological traits. In the present study, the genome sizes of 22 dictyopteran species were estimated by Feulgen image analysis densitometry and 6-diamidino-2-phenylindole (DAPI)-based flow cytometry. The haploid genome sizes (C-values) of termites (Isoptera) ranged from 0.58 to 1.90 pg, and those of Cryptocercus wood roaches (Cryptocercidae) were 1.16 to 1.32 pg. Compared to known values of other cockroaches (Blattaria) and mantids (Mantodea), these values are low. A relatively small genome size appears to be a (syn)apomorphy of Isoptera + Cryptocercus, together with their sociality. In some phylogenetic groups, genome size evolution is thought to be influenced by selective pressure on a particular trait, such as cell size or rate of development. The present results raise the possibility that genome size is influenced by selective pressures on traits associated with the evolution of sociality.

Termites are instantly recognizable mound-builders and house-eaters: their complex social lifestyles have made them incredibly successful throughout the tropics. Although known as 'white ants', they are not ants and their relationships with other insects remain unclear. Our molecular phylogenetic analyses, the most comprehensive yet attempted, show that termites are social cockroaches, no longer meriting being classified as a separate order (Isoptera) from the cockroaches (Blattodea). Instead, we propose that they should be treated as a family (Termitidae) of cockroaches. It is surprising to find that a group of wood-feeding cockroaches has evolved full sociality, as other ecologically dominant fully social insects (e.g. ants, social bees and social wasps) have evolved from solitary predatory wasps.

Isoptera are highly specialized cockroaches and are one of the few eusocial insect lineages. Cryptocercus cockroaches have appeared to many as ideal models for inference on the early evolution of termites, due to their possible phylogenetic relationship and several shared key attributes in life history. Recently, Pellens, Grandcolas, and colleagues have proposed the blaberid cockroach Parasphaeria boleiriana to be an alternative model for the early evolution in termites. We compare the usefulness of Cryptocercus and P. boleiriana as models for termite evolution. Cryptocercus and lower Isoptera (1) can both feed on comparatively recalcitrant wood, (2) have an obligate, rich and unique hypermastigid and oxymonadid fauna in the hindgut, (3) transfer these flagellates to the next generation by anal trophallaxis, (4) have social systems that involve long-lasting biparental care, and, finally, (5) are strongly suggested to be sister groups, so that the key attributes (1)–(4) appear to be homologous between the two taxa. On the other hand, P. boleiriana (1) feeds on soft, ephemeral wood sources, (2) shows no trace of the oxymonadid and hypermastigid hindgut fauna unique to Cryptocercus and lower Isoptera, nor does it have any other demonstrated obligate relationship with hindgut flagellates, (3) is likely to lack anal trophallaxis, (4) has only a short period of uniparental brood care, and (5) is phylogenetically remote from the Cryptocercus + Isoptera clade. These facts would argue against any reasonable usage of P. boleiriana as a model for the early evolution of Isoptera or even of the clade Cryptocercus + Isoptera. Cryptocercus thus remains an appropriate model-taxon-by-homology for early termite evolution. As compared to P. boleiriana, some other Blaberidae (such as the Panesthiinae Salganea) appear more useful as model-taxa-by-homoplasy for the early evolution of the Cryptocercus + Isoptera clade, as their brooding behavior is more elaborate than in P. boleiriana.