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Termite symbiotic systems: efficient bio-recycling of lignocellulose.

Termites thrive in great abundance in terrestrial ecosystems and play important roles in biorecycling of lignocellulose. Together with their microbial symbionts, they efficiently decompose lignocellulose. In so-called lower termites, a dual decomposing system, consisting of the termite's own cellulases and those of its gut protists, was elucidated at the molecular level. Higher termites degrade cellulose apparently using only their own enzymes, because of the absence of symbiotic protists. Termite gut prokaryotes efficiently support lignocellulose degradation. However, culture-independent molecular studies have revealed that the majority of these gut symbionts have not yet been cultivated, and that the gut symbiotic community shows a highly structured spatial organization. In situ localization of individual populations and their functional interactions are important to understand the nature of symbioses in the gut. In contrast to cellulose, lignin degradation does not appear to be important in the gut of wood-feeding termites. Soil-feeding termites decompose humic substances in soil at least partly, but little is known about the decomposition. Fungus-growing termites are successful in the almost complete decomposition of lignocellulose in a sophisticated cooperation with basidiomycete fungi cultivated in their nest. A detailed understanding of efficient biorecycling systems, such as that for lignocellulose, and the symbioses that provide this efficiency will benefit applied microbiology and biotechnology.

Animals↗

Influence of the diet components on the symbiotic microorganisms community in hindgut of Coptotermes formosanus Shiraki.

Artificial diet was developed for rearing of lower termites (workers) Coptotermes formosanus. C. formosanus was fed with either wood powder of Japanese red pine, cellulose, cellobiose, or glucose for 30 days. The effect of carbon sources in the diet on the structure and function of the symbiotic intestinal microbial community and on the physiological activity of C. formosanus was studied. Three symbiont protozoa, Pseudotrichonympha grassi, Holomastigotoides hartmanni, and Spirotrichonympha leidyi, were found in the hindgut of C. formosanus that fed on the diets containing carbon sources with high molecular weight (MW). However, when artificial diets containing carbohydrate with low MW were used, both P. grassi and H. hartmanni disappeared, and only few S. leidyi were alive. This suggested that both P. grassi and H. hartmanni play important roles in the digestion and utilization of carbohydrate with high MW. The denaturing gradient gel electrophoresis analysis of bacterial community in the hindgut of termites showed that the similarity between intestinal bacteria community in termites fed with diets containing high-MW carbon sources and those with low MW was only about 40%. It was apparent that changes in diets resulted to changes in intestinal microbial community, and this in turn affected cellulase activity in C. formosanus.

Animals↗

Hydrogen cyanide-producing rhizobacteria kill subterranean termite Odontotermes obesus (Rambur) by cyanide poisoning under in vitro conditions.

The subterranean termite Odontotermes obesus is an important pest of the Indian subcontinent, causing extensive damage to major agricultural crops and forest plantation trees. Control of termites by strategies employing their parasites has limitations because they have evolved a complex social structure, immune responses, and adaptive behavior toward pathogen-infected individuals. Nonparasitic rhizobacteria that produce harmful metabolites might facilitate the biocontrol of termites. In the present investigation, three different species of hydrogen cyanide-producing rhizobacteria were tested for their potential to kill O. obesus. The three bacterial species were found to be effective in killing the termites under in vitro conditions.

Alcaligenes↗

Metabolic physiology of the numbat (Myrmecobius fasciatus).

The numbat (Myrmecobius fasciatus) is unique amongst marsupials as it is exclusively diurnal, feeds only on termites and is semi-fossorial. This study examines the thermal and metabolic physiology of the numbat to determine if its physiology reflects its phylogeny, diet and semi-fossorial habit. Numbats (mean adult body mass 552 g) were able to regulate body temperature at ambient temperatures of 15-30 degrees C, with a body temperature at thermoneutrality (30 degrees C) of 34.1 degrees C. The thermoneutral body temperature was not significantly different from that predicted for an equivalent-sized marsupial. Basal metabolic rate, measured at 30 degrees C, was 0.389 +/- 0.025 ml O(2) g(-1) h(-1), and was slightly but not significantly lower at 82.5% of that predicted for a typical marsupial of equivalent body mass. Metabolic rate increased with decreasing ambient temperatures below 30 degrees C. Patterns of metabolic cycling observed for completely inactive numbats at ambient temperatures below 30 degrees C are likely to be related to sleep phase. Wet thermal conductance of 1.94 J g(-1) h(-1) degrees C(-1) (at 30 degrees C) was 131% of that predicted for a marsupial. Evaporative water loss of the numbat remained constant below the thermoneutral zone (<30 degrees C) at approximately 0.6 ml g(-1) h(-1), only 47.4% of that predicted for a marsupial. It increased to 1.01 +/- 0.16 ml g(-1) h(-1) at an ambient temperature of 32.5 degrees C. The thermal and metabolic physiology of the numbat is generally similar to that expected for other marsupials, and is also comparable to that of termitivorous placental mammals. Thus the reduction in body temperature and basal metabolic rate of placental termitivores is a "marsupial-like" low energy turnover physiology, and the numbat being a marsupial already has an appropriate physiology to survive exclusively on a low energy diet of termites.

Animals↗

The symbiotic fauna of the African termite Hodotermes mossambicus identification of four flagellate species of the genera Spironympha, Trichomonoides and Retortamonas.

Two Spironympha species were described by light immunofluorescence and electron microscopy. Spironympha fibrosa n. sp. has a narrow columella, several axostylar fibres and original striated myoneme-like fibres attached to the last basal body of each flagellar line. Spironympha simplex n. sp. is small and has non-striated microfibrils attached to the basal body section and an axostyle composed of two fibres. Trichomonas termitidis, as reported by Dogiel (Researches on parasitic protozoa from the intestine of termites. II. Lophomonadidae. Sci Res Zool Exped Brit E Africa made by Prof. V. Dogiel and I. Sokolow in 1914 10:20-35, 1917), has the features of Trichomonoides trypanoides, as reported by Brugerolle and Bordereau (Eur J Protistol 40:163-174, 2004). Retortamonas hodotermitis n. sp. shares the characteristics of insect retortamonads, as reported by Brugerolle (Protistologica 8:233-240, 1976), but does not correspond to any species described to date. The species identification differs from that of described in Dogiel (Russkii Arkhiv Protistologii 1:172-234, 1922).

Animals↗

Vertical stratification of the termite assemblage in a neotropical rainforest.

The importance of termites as decomposers in tropical forests has long been recognized. Studies on the richness and diversity of termite species and their ecological function have flourished in more recent times, but these have been mostly conducted in a thin stratum within a standing man's reach. Our aims were to evaluate the specific richness and composition of the termite assemblage in the canopy of a tropical rainforest and to determine its originality with respect to the sympatric ground-level fauna. We conducted systematic searches for canopy termites, together with conventional sampling of the sympatric ground-level fauna, in the San Lorenzo forest, Panama. We hypothesized that (1) the canopy accommodates two categories of wood-feeding termites (long-distance foragers and small-colony "one-piece" species) and possibly soil-feeders in suspended soil-like habitats; (2) due to the abundance of soil-feeders, the overall diversity of the ground fauna is higher than that of the canopy; (3) differences in microclimate and resource accessibility favour vertical stratification among wood-feeders. Sixty-three canopy samples yielded ten species of termites, all wood-feeders. Five of these were not found at ground level, although a total of 243 ground samples were collected, representing 29 species. In addition to long-distance foragers (Microcerotermes and Nasutitermes spp.) and small-colony termites (mostly Kalotermitidae), the canopy fauna included Termes hispaniolae, a wood-feeding Termitidae from an allegedly soil-feeding genus, living in large dead branches. Soil-feeders were absent from the canopy, probably because large epiphytes were scarce. As predicted, the ground fauna was much richer than that of the canopy, but the species richness of both habitats was similar when only wood-feeders were considered. Vertical stratification was strongly marked among wood-feeders, as all common species, apart from the arboreal-nesting Microcerotermes arboreus, could unequivocally be assigned to either a ground or a canopy group. The canopy, therefore, contributes significantly to the total species richness of the termite assemblage, and the diversity, abundance and ecological importance of canopy termites in tropical rainforests may be higher than previously recognized.

Animals↗

Devescovinid features, a remarkable surface cytoskeleton, and epibiotic bacteria revisited in Mixotricha paradoxa, a parabasalid flagellate.

This work reports on the flagellate systematics and phylogeny, cytoskeleton, prokaryote-eukaryote cell junction organisation, and epibiotic bacteria identification. It confirms the pioneer 1964 study on Mixotricha paradoxa and supplies new information. Mixotricha paradoxa has a cresta structure specific to devescovinid parabasalid flagellates, a slightly modified recurrent flagellum, and an axostylar tube containing two lamina-shaped parabasal fibres. However, many parabasal profiles are distributed throughout the cell body. There is a conspicuous cortical microfibrillar network whose strands are related to cell junction structures subjacent to epibiotic bacteria. The supposed actin composition of this network could not be demonstrated with anti-actin antibodies or phalloidin labelling. Four types of epibiotic bacteria were described. Bacillus-shaped bacteria with a Gram-negative organisation are nested in alternate rows on most of the surface of the protozoon. They induce a striated calyxlike junction structure beneath the adhesion zone linked to the cortical microfibrillar network. Slender spirochetes are attached by one differentiated end to the plasma membrane of the protozoon, forming knobs on the cell surface. Two very similar long rod-shaped bacteria are also attached on the knobs of the plasma membrane. A large spirochete attributed to the genus Canaleparolina is also attached to the protozoon. Observations on epibiotic bacteria and of their attachments are compared with several described epibiotic bacteria of symbiotic protozoa and with the results of the molecular identification of the epibiotic bacteria of M. paradoxa.

Animals↗

Comparison of bacterial communities in the alkaline gut segment among various species of higher termites.

The first proctodeal (P1) segment in the hindgut of certain higher termites shows high alkalinity. We examined the bacterial diversity of the alkaline P1 gut segments of four species of higher termites by T-RFLP and phylogenetic analyses based on PCR-amplified 16S rRNA genes. The bacterial community of the P1 segment was apparently different from that of the whole gut in each termite. Sequence analysis revealed that Firmicutes (Clostridia and Bacilli) were dominant in the P1 segments of all four termites; however, the phylogenetic compositions varied among the termites. Although some of the P1 segment-derived sequences were related to the sequences previously reported from the alkaline digestive tracts of other insects, most of them formed phylogenetic clusters unique to termites. Such "termite P1 clusters" were distantly related to known bacterial species as well as to sequences reported from alkaline environments in nature. We successfully obtained enrichment cultures of Clostridia- and Bacilli-related bacteria, including putative novel species under anaerobic alkaline conditions from the termite guts. Our results suggest that the alkaline gut region of termites harbors unique bacterial lineages and are expected to be a rich reservoir of novel alkaliphiles yet to be cultivated.

Animals↗

Molecular phylogenetic identification of the intestinal anaerobic microbial community in the hindgut of the termite, Reticulitermes speratus, without cultivation.

A termite maintains an anaerobic microbial community in its hindgut, which seems to be the minimum size of an anaerobic habitat. This microbial community consists of bacteria and various anaerobic flagellates, and it is established that termites are totally dependent on the microbes for the utilization of their food. The molecular phylogenetic diversity of the intestinal microflora of a lower termite, Reticulitermes speratus, was examined by a strategy that does not rely on cultivation of the resident microorganisms. Small subunit ribosomal RNA (ssrRNA) genes were directly amplified from the mixed-population DNA of the termite gut by polymerase chain reaction (PCR) and clonally isolated. Most sequenced clones were phylogenetically affiliated with the four major groups of the domain Bacteria: the Proteobacteria group, the Spirochete group, the Bacteroides group, and the Low G + C gram-positive bacteria. The 16S rRNA sequence data show that the majority of the intestinal microflora of the termite consists of new species that are yet to be cultured. The phylogeny of a symbiotic methanogen inhabiting the gut of a lower termite (R. speratus) was analyzed without cultivation. The nucleotide sequence of the ssrDNA and the predicted amino acid sequence of the mcrA product were compared with those of the known methanogens. Both comparisons indicated that the termite symbiotic methanogen belonged to the order Methanobacteriales but was distinct from the known members of this order. The diversity of nitrogen-fixing organisms was also investigated without culturing the resident microorganisms. Fragments of the nifH gene, which encodes the dinitrogenase reductase, were directly amplified from the mixed-population DNA of the termite gut and were clonally isolated. The phylogenetic analysis of the nifH amino acid sequences showed that there was a remarkable diversity of nitrogenase genes in the termite gut. The molecular phylogeny of a symbiotic hypermastigote Trichonympha agilis (class Parabasalia; order Hypermastigida) in the hindgut of R. speratus was also examined by the same strategy. The whole-cell hybridization experiments indicated that the sequence originated from a large hypermastigote in the termite hindgut, Trichonympha agilis. According to the phylogenetic trees constructed, the hypermastigote represented one of the deepest branches of eukaryotes. The hypermastigote along with members of the order Trichomonadida formed a monophyletic lineage, indicating that the hypermastigote and trichomonads shared a recent common ancestry.

Animals↗

Diverse genes of cellulase homologues of glycosyl hydrolase family 45 from the symbiotic protists in the hindgut of the termite Reticulitermes speratus.

Diverse genes encoding cellulase homologues belonging to glycosyl hydrolase family 45 were identified from the symbiotic protists in the hindgut of the termite Reticulitermes speratus through the use of consensus PCR and the screening of a cDNA library. Fifteen full-length cDNA clones were isolated and sequenced, which encoded polypeptides consisting of 218-221 amino acid residues showing up to 63% identity to known family 45 cellulases. The cellulase sequences of the termite symbiotic protists were phylogenetically monophyletic, showing more than 75% amino acid identity with each other. These enzymes consist of a single catalytic domain, lacking the ancillary domains found in most microbial cellulases. By whole-cell in situ hybridization using oligonucleotide probes specific for regions conserved in some of the sequences, the origin of the genes was identified as symbiotic hypermastigote protists. The presence of diverse cellulase homologues suggests that symbiotic protists of termites may be rich reservoirs of novel cellulase sequences.

Amino Acid Sequence↗

Speciation of termite gut protists: the role of bacterial symbionts.

At least 12 termite gut protists have been named because of their bacterial symbionts. Dozens more species are diagnosed by epi- and endosymbionts and more still have regular bacterial associations referred to in their species description. Molecular systematic studies have begun to identify these bacteria, but the ecological relations with their protist bionts are still unknown. Recent findings of acetogenic spirochetes in termite guts may explain the peculiar arrangement of spirochetes on some of these protists. Other bacteria function as motility or chemotactic symbionts of these protists. The size and shape of the parabasal body, a Golgi complex, are morphological characters of the Parabasalia (trichomonads, hypermastigids) that may be influenced by regular, heritable epi- and endosymbiotic bacteria.

Animals↗

Diet-mediated inter-colonial aggression in the formosan subterranean termite Coptotermes formosanus.

In most social insects, intercolonial and interspecific aggression are expressions of territoriality. In termites, cuticular hydrocarbons (CHCs) have been extensively studied for their role in nestmate recognition and aggressive discrimination of nonnest-mates. More recently, molecular genetic techniques have made it possible to determine relatedness between colonies and to investigate the influence of genetics on aggression. In the Formosan subterranean termite, Coptotermes formosanus, however, the role of CHCs and genetic relatedness in inter-colony aggression has been ambiguous, suggesting the involvement of additional factors in nest-mate recognition. In this study we assess the range of aggression in this termite species and characterize the influence of genetic relatedness, CHC profiles and diet on aggression levels. We collected four colonies of C. formosanus, feeding either on bald cypress or birch, from three locations in Louisiana. Inter-colony aggression ranged from low to high. Differences in CHC profiles, as well as genetic distances between colonies determined by using microsatellite DNA markers, showed no significant correlation with aggression. However, termite diet (host tree) played a significant role in determining the level of aggression. Thus, two distantly related colonies, each feeding on different diets, showed high aggression that significantly diminished if they were fed on the same wood in the laboratory (spruce). Using headspace solid phase microextraction, we found three compounds from workers fed on birch that were absent in workers fed on spruce. Such diet-derived chemicals may be involved in the complex determination of nest-mate recognition in C. formosanus.

Aggression↗

Hydroquinone is not a phagostimulant for the Formosan subterranean termite.

It has been suggested that hydroquinone found in the labial glands of a number of termite species acts as a primary phagostimulating factor. We tested hydroquinone as a phagostimulant using workers from three colonies of the Formosan subterranean termite, Coptotermes formosanus, under both laboratory and field conditions. Hydroquinone at concentrations ranging from ca. 0.002-20.0 ng/cm2 did not increase visitation by C. formosanus workers to treated over control filter papers, and was actually repellent at a 20 ng/cm2 dose. No phagostimulant response to hydroquinone was observed in two colonies. In the third, there was a significant increase in feeding on filter paper treated with a 2 ng/cm2 dose, but was significantly lower at a 20 ng/cm2 dose. Furthermore, sand treated with a gradient of hydroquinone, did not evoke increased tunneling activity compared with controls. GC-MS analysis of C. formosanus workers indicated that hydroquinone was present at an average of 41 pg/worker. It was also determined that within one week about 11% hydroquinone in aqueous solution oxidized to 1,4-benzoquinone. Our findings indicate that hydroquinone alone does not act as a phagostimulant but instead may act as a repellent at higher concentrations. The attractant/arrestant of the Formosan termite may have multiple components of which hydroquinone, at low doses, could be one.

Animals↗

Interspecific variation in defense secretions of Malaysian termites from the genus Bulbitermes.

The chemicals of the defense secretions of Malaysian Bulbitermes, B. singaporensis, B. germanus, B. sarawakensis, and Bulbitermes sp. B, show that B. singaporensis is distinct from the other species, which are themselves closely related; the genetic distance between B. singaporensis and B. germanus is 0.71. B. singaporensis contains tetracyclic kempane, and B. germanus and B. sarawakensis contain tricyclic trinervitene; Bulbitermes sp. B contains a mixture of kempane and trinervitene. The mono- and diterpenoid compositions are species-specific.

Animals↗

Role of (3Z,6Z,8E)-dodecatrien-1-ol in trail following, feeding, and mating behavior of Reticulitermes hesperus.

Trail pheromones mediate communication among western subterranean termites, Reticulitermes hesperus Banks. Repetitive passages of >or=28 termites were required to establish a pheromone trail and trails needed to be reinforced because they lasted <48 hr. The minimal threshold concentration for inducing responses from termite workers and secondary reproductives was between 0.01 and 0.1 fg/cm of (3Z,6Z,8E)-dodecatrien-1-ol (henceforth, dodecatrienol). Workers showed optimal trail-following behavior to dodecatrienol at a concentration of 10 fg/cm. Trails with concentrations >10 pg/cm were repellent to workers. Workers did not detect pheromone gradients, responding equally to increasing or decreasing gradients of dodecatrienol, and termite workers were not able to differentiate between different concentrations of dodecatrienol. Termites preferred dodecatrienol trails to 2-phenoxyethanol trails. Antennae played a key role in trail pheromone perception. Dodecatrienol acted as an arrestant for worker termites (10 fg/cm2) and male alates (5 ng/cm2), whereas sternal gland extracts from females attracted male alates. Workers and alates, upon contact with filter paper disks treated with higher doses (10 fg/cm2 and 5 ng/cm2, respectively) of dodecatrienol, were highly excited (increased antennation and palpation) and repeatedly returned to the treated disks. Dodecatrienol did not act as a phagostimulant when offered on a paper towel disk. Reticulitermes hesperus is highly responsive to dodecatrienol, and it may play an important role in orientation of workers and alates.

Animals↗

Solution structure of termite-derived antimicrobial peptide, spinigerin, as determined in SDS micelle by NMR spectroscopy.

Spinigerin is a linear antibacterial peptide derived from a termite insect. It consists of 25 amino acids and is devoid of cysteines. Spinigerin displays good lytic activities against Gram-positive and Gram-negative bacteria, but has no hemolytic activities against human erythrocytes. In this study, we present a three-dimensional solution structure of spinigerin in SDS micelles. According to CD data spinigerin has an alpha-helical conformation in the presence of TFE, DPC micelles, and SDS micelles. The three-dimensional structure of spinigerin as determined by NMR spectroscopy contains a stable alpha-helix from Lys4 to Thr23. Spinigerin (4-21), an 18-residue fragment from Lys4 to Leu21, contains a similar content of alpha-helical structure compared to native spinigerin and was found to retain antibacterial activity, too. Therefore, this alpha-helical structure and the strong electrostatic attraction between four Lys and three Arg residues in spinigerin and the negatively charged polar head groups of the phospholipids on the membrane surface play important roles in disrupting membrane and subsequent cell death.

Amino Acid Sequence↗

Tool-use and tool-making by captive, group-living orangutans (Pongo pygmaeus abelii) at an artificial termite mound.

The present study examined the use and making of tools to obtain foodstuffs in artificial-mound holes by five captive, group-living Sumatran orangutans (Pongo pygmaeus abelii). Three adult orangutans frequently stripped leaves and twigs from a branch provided (tool-making), and then inserted the tool into a hole to obtain foodstuffs (tool-using). A 5-year-old female juvenile usually used the tools that adult orangutans had previously used, but rarely made tools herself. A 2-year-old male infant did not use any tools. The adult orangutans tend to leave one to several leaves at the top of the branch than to leave many leaves on the branch or to strip all leaves. It seemed likely that tools with appropriate leaves are easier to insert into holes and obtain more foodstuffs, compared with branches with many leaves or sticks without any leaves. When the orangutans were unable to insert a tool into a hole, they usually modified the tool and/or changed their tool-using technique, such as changing how they grasped the tool. These findings are discussed from the perspectives of the orangutan's behavioral flexibility regarding tool-use skills and hierarchical organization in food-processing techniques.

Animals↗

Preliminary evaluation of fungicidal and termiticidal activities of filtrates from biomass slurry fuel production.

Biomass slurry fuel (BSF) production has recently been developed as a natural energy for the conversion of solid biomass into fuel. In addition to using fuel, filtrates from BSF production may also serve a chemical source with several organic compounds. There is an increasing interest in the research and application of biomass-based filtrates. In this study, fungicidal and termiticidal properties of filtrates from BSF production using sugi (Cryptomeria japonica) and acacia (Acacia mangium) wood were evaluated in laboratory decay and termite resistance tests. Wood blocks treated with the filtrates showed increased resistance against brown-rot fungus, Fomitopsis palustris. However the filtrates from sugi wood processed at 270 degrees C which contained less phenolic compounds than the other filtrates were effective against white-rot fungus, Trametes versicolor. Phenolic compounds of filtrates seemed to play a role in the decay resistance tests however the filtrates did not increase the durability of the wood blocks against subterranean termites Coptotermes formosanus. Despite high acetic and lactic acid content of the filtrates, vanillin content of the filtrates may have served as an additional food source and promoted termite attack. It can be concluded that filtrates with phenolic compounds from lignin degradation during BSF production can be considered for targeted inhibition of brown-rot.

Animals↗