Secretory oviducts contribute to the high egg-laying rate of physogastric termite queens (Isoptera: Termitidae).

Physogastric termite queens are characterized by a notorious enlargement of the abdomen triggered by an equal development of the ovaries. Other physogastry-related modifications have been reported on the fat body, cuticle, midgut, tracheal system, and hemolymph. Surprisingly, modifications on the lateral oviducts of these females, important sites for ovulation and egg transport, have received little attention. Here we took advantage of the high fecundity of physogastric queens in three termitid species to evaluate ovary development and also to compare the morphophysiological features of the lateral oviducts between early-mated and physogastric queens of Cornitermes cumulans. Older queens show well-developed ovaries, with numerous ovarioles connected to the lateral oviducts through pedicels. At these sites, several corpora lutea were observed, residual follicle cells from previous ovulation events. Such features were absent among early-mated queens and reflect then the maturity and ageing of the queens. Histological and histochemical analyses indicated that secretory activity of the lateral oviducts was also restricted to physogastric queens, in which proteins, but not polysaccharides, are secreted into the oviduct lumen. The likely function of these proteins, based on previous studies, is to lubricate the lateral oviducts and stimulate muscular contractions to the egg transport. The physogastry of termite queens is a notorious feature, characterized by several body modifications, especially concerning the ovaries. Our results shed light on the physogastry-related changes in the lateral oviducts of termite queens, as their increasing secretory activity is in agreement with the high number of eggs produced and transporting through these structures. Thus, such changes correspond to an important step allowing the high egg-laying rate shown by physogastric termite queens.

First Neotropical record of the association between brown sclerotium-forming fungi and termite eggs in a nest of Coptotermes gestroi (Blattaria, Isoptera, Rhinotermitidae).

Insects and fungi are abundant in many environments, in which facultative and/or obligate associations involving these groups have been established during evolution. In termites, mutualism with fungi is well reported for some termite lineages (e.g., Macrotermitinae). Within some subterranean termite species (Rhinotermitidae), egg-mimicking fungi, also referred to as "termite "balls", are often harbored inside the nest, mixed to the egg piles. Such interaction seems to be advantageous for both partners since the fungi are protected inside the nest while they may serve as an additional food source and also provide cellulases which may be incorporated into the termite digestive process. Although such mutualism has been reported for seven species of Reticulitermes and Coptotermes formosanus, all the samplings were restricted to temperate regions. Here, we provide the first Neotropical record of this termite-fungus association, and the first report for Coptotermes gestroi. The morphological characters of the "termite balls" observed in a C. gestroi nest resemble those already reported for Reticulitermes spp. and the congeneric species C. formosanus. They include a color ranging from light to dark brown, spherical shape, and a reduced diameter (0.23-0.34 mm). Our findings provide new insights into the geographical distribution of the association between termites and sclerotium-forming fungi. Future genetic analyses will be valuable aiming to identify the egg-mimicking fungi associated with C. gestroi and shed light on the evolution of this fascinating symbiosis.

Comparative reproductive biology of pre-, imaginal, and neotenic castes of the Asian termite Coptotermes gestroi (Blattaria, Isoptera, Rhinotermitidae).

The Asian subterranean termite Coptotermes gestroi is a worldwide structural pest, although its reproductive biology has been poorly investigated due to a cryptic habit and occurrence of polycalic nests. In this study, we investigated ovarian development and oogenesis in different-aged females of C. gestroi: fourth-instar nymphs, non-functional neotenics, alates, and functional queens. We show that the ovaries develop gradually according to their age and functionality, as younger individuals possess immature oocytes, whereas alates and functional queens always undergo vitellogenesis. Oocytes were classified into previtellogenic (stages I, II, and III) or vitellogenic (stages IV, V, and VI). Ovary development varied among non-functional neotenics, and a rapid differentiation and/or the presence of primary reproductives are believed to influence such a maturation. Immature oocyte stages were shared between fourth-instar nymphs and neotenics. These characteristics, together with other neotenic features (wing buds, body pigmentation, and eye color), should be evaluated in detail aiming to clarify which nymphal instars differentiate into secondary reproductives. Oogenesis was not uniform among alate females, and cross-sectional area of terminal oocytes was significantly smaller in alates when compared to functional queens, suggesting different degrees of maturation in swarming individuals. Functional queens always had mature terminal oocytes (stage VI). Ovariole number and oocyte maturation in C. gestroi relies on several factors and may therefore differ among individuals of the same caste. Future studies should take into account these reproductive features to evaluate how they impact colony development.

Salivary glands in workers of Ruptitermes spp. (Blattaria, Isoptera, Termitidae, Apicotermitinae): a morphological and preoteomic approach.

Salivary glands are omnipresent in termites and occur in all developmental stages and castes. They function to produce, store, and secrete compounds, ranging from a feeding function to defensive mechanisms. Here, we provide a complete morphological overview of the salivary glands in the soldierless species Ruptitermes reconditus and R. xanthochiton, and the first proteomic profile of the salivary glands in a Neotropical Apicotermitinae representative, R. reconditus. Salivary glands from both species were composed of several acini, roughly spherical structures composed of two types of central cells (type I and II) and peripheral parietal cells, as well as transporting ducts and two salivary reservoirs. Central cells were richly supplied with electron-lucent secretory vesicles and rough endoplasmic reticulum, a feature of protein-secreting cells. Parietal cells of Ruptitermes spp. had conspicuous characteristics such as electron-lucent secretory vesicles surrounded by mitochondria and well-developed microvilli. Moreover, different individuals showed variation in the secretory cycle of salivary acini, which may be related to polyethism. Ultrastructural analysis evidenced a high synthesis of secretion and also the occurrence of lysosomes and autophagic structures in central cells. Proteomic analysis of the salivary glands revealed 483 proteins divided into functional groups, highlighting toxins/defensins and compounds related to alarm communication and colony asepsis. Soldierless termites are quite successful, especially due to morphological adaptations of the workers, including unknown modifications of exocrine glands. Thus, according to our morphological and proteomic findings, we discuss the potential roles of the salivary gland secretion in different social aspects of the sampled species.

Involvement of the Salivary Glands in the Suicidal Defensive Behavior of Workers in Neocapritermes opacus (Blattaria, Isoptera, Termitidae).

Suicidal behavior in termite workers is an extreme defensive strategy, probably a consequence of having a low number of soldiers available in the colony and there being high predation from enemies. We investigated the suicidal mechanism in workers of the Neotropical termite Neocapritermes opacus, which involves salivary gland autothysis followed by body cuticle rupture and the release of a defensive secretion. Autothysis was triggered by a physical stimulus such as a soldier bite that causes the protrusion of the salivary acini, burst reservoirs, and foregut. Histochemical and ultrastructural analyses showed salivary acini composed of peripheral parietal cells and two types of central cells, types I and II. Type I cells are filled with large electron-lucent secretory vesicles, which reacted positively to bromophenol blue and xylidine-Ponceau tests, indicating the occurrence of proteins. Type II cells are elongated and display smaller apical secretory vesicles. Parietal cells present an intracellular canaliculus with dense microvilli and cytoplasm rich in mitochondria and large electron-dense vesicles, which may participate in the self-destructive mechanism. Worker suicidal behavior was previously reported for N. taracua and N. braziliensis. N. opacus is a new species in which a salivary weapon has been developed and factors contributing to this altruistic response are discussed.

Worker Defensive Behavior Associated with Toxins in the Neotropical Termite Neocapritermes braziliensis (Blattaria, Isoptera, Termitidae, Termitinae).

Termite societies are abundant in the tropics, and are therefore exposed to multiple enemies and predators, especially during foraging activity. Soldiers constitute a specialized defensive caste, although workers also participate in this process, and even display suicidal behavior, which is the case with the species Neocapritermes braziliensis. Here we describe the morphology, mechanisms of action, and proteomics of the salivary weapon in workers of this species, which due to the autothysis of the salivary glands causes their body rupture, in turn releasing a defensive secretion, observed during aggressiveness bioassays. Salivary glands are paired, composed of two translucent reservoirs, ducts and a set of multicellular acini. Histological and ultrastructural techniques showed that acini are composed of two types of central cells, and small parietal cells located in the acinar periphery. Type I central cells were abundant and filled with a large amount of secretion, while type II central cells were scarce and presented smaller secretion. Parietal cells were often paired and devoid of secretion. The gel-free proteomic approach (shotgun) followed by mass spectrometry revealed 235 proteins in the defensive secretion, which were classified into functional groups: (i) toxins and defensins, (ii) folding/conformation and post-translational modifications, (iii) salivary gland detoxification, (iv) housekeeping proteins and (v) uncharacterized and hypothetical proteins. We highlight the occurrence of neurotoxins previously identified in arachnid venoms, which are novelties for termite biology, and contribute to the knowledge regarding the defense strategies developed by termite species from the Neotropical region.

Location, morphology and function of nephrocytes in termites.

Insect nephrocytes are cells bathed in hemolymph and considered to have an excretory function. These cells have ambiguous nomenclature and are understudied in termites. This study is the first report on the occurrence, morphology and function of nephrocytes in different termite castes. Cytological characteristics in specific developmental stages and castes enable physiological functions to be inferred. Perforate diaphragms indicate a role in filtration, while the extensive peripheral invaginations of the cell membrane suggest active endocytosis. A sequence of morphologies in putative digestive vacuoles infers a lysosomal system and the occurrence of phosphatases suggests a function involving detoxification of substances sequestered from hemolymph. Pericardical nephrocytes took up the dye trypan blue injected in live termites, suggesting their activity connected to the filtration of the hemolymph. Additionally, histochemical tests showed the existence of stored proteins in their cytoplasm. These cells present a well-developed Golgi apparatus and abundant rough endoplasmic reticulum, consistent with protein synthesis. This study highlights the importance of nephrocytes in Isoptera and opens perspectives for further research of these cells.

Evaluation of growth and foraging in laboratory colonies of Coptotermes gestroi (Isoptera, Rhinotermitidae).

Coptotermes gestroi is an exotic species in Brazil that forages on several food items at the same time. Different laboratory tests were performed with 6-year-old colonies of this termite in order to follow the development and displacement of these colonies, considering which castes and instars were transferred or moved from their original chamber. Two bioassays were carried out, one with similar alternative foods (Pinus sp. sawdust), and the other with different alternative foods (large block of Pinus taeda and small block of Pinus elliottii). The colonies of C. gestroi did not show preferences in the allocation of foragers when they had alternative similar foods. However, they were found in the chamber that contained the small block of P. elliottii, when termites had the alternative of different blocks of Pinus, indicating an environment most favorable for the nucleus of the colony (royal cup and brood). Many factors may have influenced this choice as more nutritive alimentary resource for brood or the presentation of the food, which allowed better colony accommodation. C. gestroi did not have difficulties of migration from one chamber to another and this characteristic may be a risk factor in urban infestations of this severe pest.

The fat body of termites: functions and stored materials.

The functions of the fat body in the different castes of termites, and accumulation of energy reserves, proteins and urates within this organ, are reviewed. The termite fat body is involved in multiple metabolic activities, including recycling of nitrogen. Termite fat body showed three different types of cells: adipocytes, urocytes and mycetocytes, the latter restricted to the species Mastotermes darwiniensis. Adipocytes synthesize and store lipids, glycogen and several proteins. These cells also elaborate important peptides, including some that act in immune processes. Urocytes are responsible for the storage of spherocrystals of urates, which vary quantitatively among the termite castes. The different metabolic functions of the fat body in the several castes and stages of termites are associated with specific adipocyte morphologies. The synthesis and storage of different compounds modify the structure of the fat body; this differentiation is coordinated by hormones involved with molting and reproductive cycles.