Characterization of Nasutitermes globiceps (Isoptera: Termitidae) Esterases

Esterases of Nasutitermes globiceps termites which occur on the Upper Paraná River floodplain (Brazil) were characterized. The electrophoretic pattern of the termite esterases Nasutitermes globiceps was obtained by starch gel electrophoresis. Six esterase activity zones were obtained and numbered, with esterase-1 being the most anodall one and esterase-6 the most cathodal one. Esterase-2 was detected only with substrates derived from the 4-methylumbelliferyl radical. The esterases of N. globiceps present wide substrate specificity, having been observed with substrates derived from alpha-naphthyl (acetate, propionate, and butyrate) and beta-naphthyl (acetate, butyrate) and from 4-methylumbelliferyl (acetate, propionate and butyrate). Esterase-6 is a caste-specific enzyme detected in soldiers. Only esterases 1, 3 and 5 were detected in nymphs. No genetic polymorphism has been detected thus far in the esterases of Nasutitermes globiceps. This study suggests that allozyme variation can be explored to understand Nasutitermes social structure.     

Queen-specific volatile in a higher termite Nasutitermes takasagoensis (Isoptera: Termitidae)

In social insect colonies, queen-produced pheromones have important functions in social regulation. These substances influence the behavior and physiology of colony members. A queen-produced volatile that inhibits differentiation of new neotenic reproductives was recently identified in the lower termite Reticulitermes speratus. However, there are no known queen-specific volatiles of this type in any other termite species. Here, we report volatile compounds emitted by live queens of the higher termite Nasutitermes takasagoensis. We used headspace gas chromatography mass spectroscopy (HS GC-MS) to analyze volatiles emitted by live primary queens, workers, soldiers, alates, and eggs collected in a Japanese subtropical forest. Among 14 detected compounds, 7 were soldier-specific, 1 was alate-specific, 1 was egg-specific, and 1 was queen-specific. The queen-specific volatile was phenylethanol, which is different than the compound identified in R. speratus. The identification of this queen-specific volatile is the first step in determining its functions in higher termite social regulation. Comparisons of queen pheromone substances regulating caste differentiation among various termite taxa will contribute to a better understanding of the evolution of social systems in termites.     

Purified Cladonia verticillaris lichen lectin: Insecticidal activity on Nasutitermes corniger (Isoptera: Termitidae)

Cladonia verticillaris lichen lectin (ClaveLL) was isolated through Sephadex G-100 gel filtration chromatography and characterized as a pure lectin through ÄKTA-FPLC and HPLC systems. The lichen extract (LE), protein fraction (F₁) and ClaveLL were assayed to evaluate their potential insecticidal and/or repellence activities on termite Nasutitermes corniger. LE, F₁ and ClaveLL were evaluated for hemagglutinating activity (HA), protein concentration and presence of secondary metabolites; preparations and active ClaveLL, free of secondary metabolites, were able to induce termite mortality. ClaveLL LC₅₀ values after 10 days for workers and soldiers were 0.196 and 0.5 mg ml^?1, respectively. C. verticillaris preparations are potential tools for researches involving control of termites (or other insects) of economic interest to wooden industry or agriculture as well as preservation of plant species that are targets of termites or other plagues.     

Population structure and genetic diversity in insular populations of Nasutitermes takasagoensis (Isoptera: Termitidae) analyzed by AFLP markers

Dispersal ability and degree of inbreeding in a population can indirectly be assessed using genetic markers. In general, it was suggested that winged termites are not able to fly distances greater than several hundred meters. Here, amplified fragment length polymorphism (AFLP) was used to analyze genetic diversity, population substructure, and gene flow among insular populations of the termite Nasutitermes takasagoensis (Isoptera: Termitidae) in the Yaeyama Islands, Okinawa, Japan. Samples were collected from 77 nests on seven islands of the Yaeyama Group. Using three primer combinations a total of 155 bands were generated with 78 (50%) polymorphic bands. Genetic distance and G(st) values among insular populations were calculated. Relatively high genetic diversity and low values of G (st), suggest there is moderate subpopulation structure. Based on these results, we discussed two possibilities; first, winged termites are able to fly over distances of several kilometers, and second, these results were obtained because insular populations share a recent common origin.     

A sex-linked ring quadrivalent in Termitidae (Isoptera)

A sex-linked ring of four multiple has been identified at male meiosis in 21 African species of Termitidae. Since this ring occurs only in the males it is clear that it represents a multiple sex chromosome system of the X₁X₂Y₁Y₂X₁X₁X₂X₂ type.     

Forces affecting water imbibition inMacrotermes workers (Termitidae,Isoptera)

Summary Observations of putative drinking behaviour at soil surfaces by major workers ofMacrotermes subhyalinus andM. bellicosus suggested that unidirectionally arranged trichomes on the anterior hypopharynx exert capillarity on water in soil pores. To test this hypothesis thirsty major workers were placed on soil of relatively low water content. From the soil types tested we determined soil-moisture retention curves which represent graphically the water content of the soil and its corresponding capillary potential. The termites were able to obtain water from different types of clay soil against potentials in the range – 1.6 to – 1.9 bar. On sandy soil the capillary potential was found to be much higher (– 0.18 bar), i.e. closer to zero. Sandy soil has a very unfavourable water retention capacity, i.e. its water content is very low at low capillary potential, whereas clay soil retains much more water at low capillary potential. The ability of the termites to gain water from soils with relatively low water content is an important adaptive feature for survival in semi-arid areas.     

Growth and demography ofCubitermes speciosus mounds (Isoptera,Termitidae)

The growth and demography ofCubitermes speciosus mounds were studied in an equatorial forest ecosystem near Kisangani, Zaïre. The mounds were censused and measured on eight occasions from June 1985 to January 1993 on 25 plots of 100 m².Mounds were found to grow in two ways: either they were enlarged sporadically, leading to a correlation between age and size; or they were no longer enlarged after a 2-year period of initial growth, which means that both large and small mounds can be old.The number of living mounds was fairly constant: it varied between 112 and 152 ha^–1 with an average of 139 ha^–1, and about a quarter of them were renewed each year. However, a clear tendency was observed over our 7.5-year observation period: the average size of the mounds increased regularly and was 2.7 times larger in 1993 than in 1985.Monthly birth and death rates were estimated using a model of continuous growth. Life expectancy at the age of 3 months was estimated from a survivorship curve at 3.5 years.     

High gene flow between populations of Macrotermes michaelseni (Isoptera, Termitidae)

Termite alates are thought to be poor active flyers, and this should lead to considerable genetic differentiation on small spatial scales. However, using four microsatellite loci for the termite Macrotermes michaelseni we found low values of genetic differentiation (F_ST) across a spatial scale of even more than 50 km. Genetic differentiation between populations increased with spatial distance up to 50 km. Furthermore, up to this distance, the scatter around the linear regression of genetic differentiation versus spatial distance increased with spatial distance. This suggests that across such spatial distances gene flow and genetic drift are of about equal importance, and near equilibrium. Using a regional F_ST as well as the distance between populations with non-significant F_ST-values (up to 25 km), gene flow is sufficiently high so that populations may be regarded as panmictic on spatial scales of 25 to 50 km. The apparent contradiction between dispersal distances observed in the field and estimates of gene flow from genetic markers may be due to the masses of swarming alates. Assuming a leptokurtic distribution of dispersal distances, atleast some alates are expected to travel considerable distances, most likely by passive drift. Received 25 January 2005; revised 11 April 2005; accepted 26 April 2005.     

Redescription of the enigmatic genus Genuotermes Emerson (Isoptera,Termitidae, Termitinae)

The imago and soldier castes of the Neotropical Termitinae species Genuotermes spinifer Emerson are redescribed. The gut anatomy of the worker is described in detail for the first time, and morphological variations in the soldier are noted and illustrated. The known geographical distribution of Genuotermes spinifer is greatly expanded.     

Fine Structure of the Labial Gland in Macrotermes bellicosus (Isoptera,Termitidae)

The labial gland in M. bellicosus corresponds with the acinar type, and occupies the greater part of meso- and metathorax. The acini comprise three secretory cell types, in addition to the central ductule cells and the epithelial cells that make up the efferent ducts. Cell types are mainly distinguished by the size and appearance of their secretory vesicles and the extent of the microvillar contact area with the ductule cells. They probably produce a proteinaceous secretion that may contain digestive enzymes. The labial gland acini in soldiers, on the other hand, contain only one type of secretory cell, which is not comparable with any of the cell types in the worker caste. This difference is in agreement with the multifunctional role of the labial gland according to the termites' polyethism.     

Occurrence of fungi in combs of fungus-growing termites (Isoptera: Termitidae, Macrotermitinae)

Fungus-growing termites cultivate their mutualistic basidiomycete Termitomyces species on a substrate called a fungal comb. Here, the Suicide Polymerase Endonuclease Restriction (SuPER) method was adapted for the first time to a fungal study to determine the entire fungal community of fungal combs and to test whether fungi other than the symbiotic cultivar interact with termite hosts. Our molecular analyses show that although active combs are dominated by Termitomyces fungi isolated with direct Polymerase Endonuclease Restriction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE), they can also harbor some filamentous fungi and yeasts only revealed by SuPER PCR-DGGE. This is the first molecular evidence of the presence of non-Termitomyces species in active combs. However, because there is no evidence for a species-specific relationship between these fungi and termites, they are mere transient guests with no specialization in the symbiosis. It is however surprising to notice that termite-associated Xylaria strains were not isolated from active combs even though they are frequently retrieved when nests are abandoned by termites. This finding highlights the implication of fungus-growing termites in the regulation of fungi occurring within the combs and also suggests that they might not have any particular evolutionary-based association with Xylaria species.     

Behavioural differences between male and female replacement reproductives in Kalotermes flavicollis (Isoptera, Kalotermitidae)

Summary: An ethogram type study was performed on representative colonies of Kalotermes flavicollis to investigate the behavioural repertoires of replacement king and queen. Our observations suggested a sex-based behavioural specialisation in the two reproductives, affecting colony organisation and equilibrium. Interesting differences were detected in overall activity of the two reproductives, with the male showing the greatest frequency of movements in the nest, vibratory acts, and the highest rate and range of social contacts. In addition to his importance for periodical insemination of the queen, the king might also play a basic role in the social structure and dynamic development of the colony, as a "mediator" in social interactions between the queen and the other colony members.     

Divergence times in the termite genus Macrotermes (Isoptera: Termitidae)

The evolution of fungus-growing termites is supposed to have started in the African rain forests with multiple invasions of semi-arid habitats as well as multiple invasions of the Oriental region. We used sequences of the mitochondrial COII gene and Bayesian dating to investigate the time frame of the evolution of Macrotermes, an important genus of fungus-growing termites. We found that the genus Macrotermes consists of at least 6 distantly related clades. Furthermore, the COII sequences suggested some cryptic diversity within the analysed African Macrotermes species. The dates calculated with the COII data using a fossilized termite mound to calibrate the clock were in good agreement with dates calculated with COI sequences using the split between Locusta and Chortippus as calibration point which supports the consistency of the calibration points. The clades from the Oriental region dated back to the early Tertiary. These estimates of divergence times suggested that Macrotermes invaded Asia during periods with humid climates. For Africa, many speciation events predated the Pleistocene and fall in range of 6-23 million years ago. These estimates suggest that savannah-adapted African clades radiated with the spread of the semi-arid ecosystems during the Miocene. Apparently, events during the Pleistocene were of little importance for speciation within the genus Macrotermes. However, further investigations are necessary to increase the number of taxa for phylogenetic analysis.     

Isolation and characterization of microsatellite markers in Labiotermes labralis (Isoptera,Termitidae, Nasutitermitinae)

We report here on the development of six polymorphic microsatellite loci for Labiotermes labralis. This soil‐feeding species is restricted to the Neotropical rainforest and then could represent a major candidate for being a bio‐indicator of anthropic disturbance resulting in the forest fragmentation. The microsatellite loci were isolated and characterized using a microsatellite‐enriched genomic library. The primers were tested on a French Guyanese population of L. labralis, represented by the sampling of one soldier in 22 nests. The primers were also tested against nine species of the same Nasutitermitinae subfamily and were successfully amplified at five loci in Armitermes minutus.