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Termite feeding deterrent from Japanese larch wood.

Extraction of flavonoids from Japanese larch (Larix leptolepis) wood with water was carried out to prepare a termite feeding deterrent. A two-stage procedure for the extraction was adopted. The first extraction step was performed at ambient temperature (22 degrees C) and the second at elevated temperatures ranging 50-100 degrees C. The first step mainly gave a mixture of polysaccharides together with small amount of flavonoids. At the second step, the yield of extract and its chemical composition were greatly affected by the temperature. The yield of solubilised carbohydrates steadily increased with a rise in the temperature, while the overall yield of flavonoids reached its optimum at 70 degrees C. An additional increase in the temperature resulted in a decrease in the yield. Model experiments using dihydroflavonols confirmed the occurrence of oxidative dehydrogenation and/or intramolecular rearrangement during the hydrothermal treatment at higher temperatures. The crude water extracts showed strong feeding deterrent activities against the subterranean termite, Coptotermes formosanus, in a choice paper disc assay. The extracts containing flavonoids in large quantities exhibited potent termite feeding deterrent activities.

Animals↗

Isolation and characterization of enteric bacteria from the hindgut of Formosan termite.

The Formosan subterranean termite, Coptotermes formosanus Shiraki, is an aggressive, invasive termite species that has caused billions of dollars of damage across the United States for the past 50 years. Termites depend on intestinal microorganisms for cellulose digestion. Symbiotic microorganisms in the termite gut play key physiological functions such as cellulose and hemicellulose digestion, acetogenesis, hydrogenesis, methanogenesis, sulfate reduction, and nitrogen fixation. Additionally, intestinal microbes create suitable conditions for symbiotic protozoans through the production of nutrients and the maintenance of the pH and the anaerobic conditions in the gut. Although extensive research has been done on the symbiotic relationship of these termites and the microbes found in its gut, there is little information available on the role of facultative anaerobes in the gut. We isolated four enteric bacteria from the hindgut of Formosan subterranean termite, C. formosanus. All isolates were facultative anaerobes and G-. The isolates were identified as Serratia marcescens, Enterobacter aerogens, Enterobacter cloacae, and Citrobacter farmeri by using BIOLOG assay and fatty acid methyl ester analysis (FAME). Each isolate was characterized using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis and biochemical study. This is the first report on the presence of facultative microbes in termite gut. Results of this first study on facultative microbes in the termite gut indicate that the role of facultative organisms in the Formosan termite gut may be to scavenge oxygen and create anaerobic conditions for the anaerobic microorganisms, which are essential for digestion of cellulose consumed by the termite.

Acyltransferases↗

Anti-termitic activities of essential oils from coniferous trees against Coptotermes formosanus.

In this study, the anti-termitic activities of 11 essential oils from three species of coniferous tree against Coptotermes formosanus Shiraki were investigated using direct contact application. Results demonstrated that at the dosage of 10 mg/g, the heartwood and sapwood essential oils of Calocedrus macrolepis var. formosana and Cryptomeria japonica and the leaf essential oil of Chamaecyparis obtusa var. formosana had 100% mortality after 5 d of test. Among the tested essential oils, the heartwood essential oil of C. macrolepis var. formosana killed all termites after 1 d of test, with an LC(50) value of 2.6 mg/g, exhibiting the strongest termiticidal property. The termiticidal effect of heartwood essential oil was due to its toxicity and its repellent action.

Animals↗

Evaluation of thermally modified Grevillea robusta heartwood as an alternative to shortage of wood resource in Kenya: Characterisation of physicochemical properties and improvement of bio-resistance.

Heat treatment of Grevillea robusta, a tropical wood species of low natural durability, was carried-out under inert conditions to improve its decay resistance. Resistance of heat treated samples was evaluated by malt agar block tests after three months of exposure to several wood rotting fungi. Also resistance of heat treated wood against termites was tested in the laboratory and in the field. Results showed that durability against fungi and termites was greatly improved after treatment. There was a good correlation between decay resistance and mass loss due to thermal treatment. Microscopic, FTIR and (13)C MAS NMR analysis were performed to characterize wood chemical and anatomical modifications that occur after treatment to understand the reasons of the durability improvement.

Animals↗

Food encounter rate of simulated termite tunnels in heterogeneous landscapes.

The aim of this study was to explore how a heterogeneous landscape affects food encounter rate in the Formosan subterranean termite, Coptotermes formosanus Shiraki. To do this, a lattice model was formulated to simulate the tunneling structure of the termite. The model made use of minimized local rules derived from empirical data. In addition, a landscape structure was generated on a lattice space by using a neutral landscape model. Each lattice cell has a value h, representing spatially distributed property of the landscape (e.g., temperature or moisture). The heterogeneity of the landscape was characterized by a parameter, H controlling aggregation of lattice cells with higher values of h. Higher H values correspond to higher aggregation levels. The effect of the landscape heterogeneity on the encounter rate was clear in the presence of higher food density than in lower density. The effect was also enhanced by the increase of the number of primary tunnels.

Animal Feed↗

The gut flagellate community of the termite Neotermes cubanus with special reference to Staurojoenina and Trichocovina hrdyi nov. gen. nov. sp.

At least seven species of flagellates have been found to inhabit the paunch of the termite Neotermes cubanus. Staurojoenina sp. is the largest species, measuring 100-150mum in length. Three small parabasalids belong to the genera Tritrichomonas, Metadevescovina, and Foaina. A fourth small type is described as Trichocovina hrdyi nov. gen. nov. sp.; the combination of features in this flagellate, such as the presence of a costa, undulating membrane and spiralled dictyosome, does not fit to any known genus. The two oxymonad species do not possess a rostellum. One belongs to the family Polymastigidae; it has one unattached flagellum and three partially attached flagella. The second is a member of the family Pyrsonymphidae, but this one possesses ring-like surface structures, one free flagellum and three spirally attached flagella. It is the first report of a pyrsonymphid in a kalotermitid termite.

Animals↗

Atmospheric versus biological sources of polycyclic aromatic hydrocarbons (PAHs) in a tropical rain forest environment.

To distinguish between pyrogenic and biological sources of PAHs in a tropical rain forest near Manaus, Brazil, we determined the concentrations of 21 PAHs in leaves, bark, twigs, and stem wood of forest trees, dead wood, mineral topsoil, litter layer, air, and Nasutitermes termite nest compartments. Naphthalene (NAPH) was the most abundant PAH with concentrations of 35 ng m(-3) in air (>85% of the sum of 21PAHs concentration), up to 1000 microg kg(-1) in plants (>90%), 477 microg kg(-1) in litter (>90%), 32 microg kg(-1) in topsoil (>90%), and 160 microg kg(-1) (>55%) in termite nests. In plants, the concentrations of PAHs in general decreased in the order leaves > bark > twigs > stem wood. The concentrations of most low-molecular weight PAHs in leaves and bark were near equilibrium with air, but those of NAPH were up to 50 times higher. Thus, the atmosphere seemed to be the major source of all PAHs in plants except for NAPH. Additionally, phenanthrene (PHEN) had elevated concentrations in bark and twigs of Vismia cayennensis trees (12-60 microg kg(-1)), which might have produced PHEN. In the mineral soil, perylene (PERY) was more abundant than in the litter layer, probably because of in situ biological production. Nasutitermes nests had the highest concentrations of most PAHs in exterior compartments (on average 8 and 15 microg kg(-1) compared to <3 microg kg(-1) in interior parts) and high PERY concentrations in all compartments (12-86 microg kg(-1)), indicating an in situ production of PERY in the nests. Our results demonstrate that the deposition of pyrolytic PAHs from the atmosphere controls the concentrations of most PAHs. However, the occurrence of NAPH, PHEN, and PERY in plants, termite nests, and soils at elevated concentrations supports the assumption of their biological origin.

Air Pollutants↗

Screening of genes expressed in developing mandibles during soldier differentiation in the termite Hodotermopsis sjostedti.

We investigated the morphological changes accompanying soldier differentiation in the damp-wood termite Hodotermopsis sjostedti. Genes expressed in the developing mandibles, which undergo the most remarkable morphological changes during soldier differentiation, were screened using fluorescent differential display. Database searches for sequence similarities were conducted and the relative expression levels were then quantified by real-time polymerase chain reaction. Among the identified candidate genes, 12 genes were upregulated during soldier differentiation. These included genes for cuticle proteins, nucleic acid binding proteins, ribosomal proteins and actin-binding protein, which were inferred to be involved in caste-specific morphogenesis in termites.

Animals↗

Spatial distribution of bacterial phylotypes in the gut of the termite Reticulitermes speratus and the bacterial community colonizing the gut epithelium.

The bacterial community colonizing the gut wall of the termite Reticulitermes speratus was characterized without cultivation. Analysis of 16S rRNA genes after fractionation of the gut revealed that the bacterial composition on the gut wall was diverse and significantly different from that able to move unconfined in the gut fluid or physically associated with the gut protists. Actinobacteria, Firmicutes and Bacteroidetes were dominant on the gut wall, but Spirochaetes and the Termite group 1 phylum, abundant in the gut lumen, were relatively rare. A sequence-specific probe enabled the in situ detection of a rod-shaped Actinobacteria member, abundantly colonizing the gut paunch epithelium.

Animals↗

Molecular cloning and characterization of a cellulase gene from a symbiotic protist of the lower termite, Coptotermes formosanus.

The endo-beta-1,4-glucanase gene was cloned from a cDNA library constructed from the mixed population of symbiotic protists in the hindgut of the lower termite, Coptotermes formosanus, using the lambda ZAP II vector. The recombinant phage library was screened for cellulolytic activity by the Congo red staining procedure. The nucleotide sequence comprised 941 nucleotides including a polyA tail sequence and showed high sequence similarity with endoglucanase genes belonging to glycosyl hydrolase family 5. Determination of the 5' end of the cellulase gene using the 5'RACE method showed that the full-length cDNA comprised a 921-bp ORF, encoding a putative 33,620 Da protein. The organismal source of this cellulase gene was identified using PCR with gene-specific primers and whole-cell in situ hybridization as the smallest symbiotic hypermastigote protist, Spirotrichonympha leidyi. The optimal pH and temperature of the cellulase heterologously expressed in Escherichia coli were 5.8-6.0 and 70 degrees C, respectively. The Km and Vmax values on carboxymethyl cellulose (CMC) substrate were 1.90 mg/ml and 148.2 units/mg protein, respectively.

Amino Acid Sequence↗

Two hexamerin genes from the termite Reticulitermes flavipes: Sequence, expression, and proposed functions in caste regulation.

Previous molecular studies on the termite Reticulitermes flavipes have revealed that two hexamerin proteins serve an important status quo role in the regulation of juvenile hormone (JH)-dependent caste differentiation. Here, we report sequence data and other experimental evidence suggesting how these two hexamerins function in achieving caste regulation. The two hexamerin genes, named Hex-1 and Hex-2, encode highly unique sequence features relative to the 100+ other known insect hexamerins. These features include a long hydrophobic tail and prenylation motif in Hex-1, and a long hydrophilic insertion plus several putative protease cleavage sites in Hex-2. Both hexamerin genes are primarily expressed in fat body tissue, but only Hex-2 expression is substantially induced by JH. SDS-PAGE showed that the hexamerin proteins constitute a major proportion of total soluble termite protein. Also, although each protein occurs in both the membrane and soluble protein fractions, Hex-2 has stronger membrane affinity. Anti-JH antiserum specifically recognizes hemolymph-soluble Hex-1 protein, supporting that the unique prenylation site in Hex-1 facilitates covalent JH binding to the primary amino acid chain. Finally, increased ratios of Hex-2 to Hex-1 transcription occur in caste phenotypes and developmental stages that differentiate in response to rising JH titers. Two main conclusions can be taken from these studies. First, elevated ratios of Hex-2 to Hex-1 expression are associated with caste phenotypes that differentiate in response to rising JH titers (i.e., workers, presoldiers and soldiers). Second, due to their unique structural features and other observed characteristics, our findings support the hypothesis that the two hexamerins participate in the regulation of caste-differentiation by modulating JH availability.

Animals↗

Effects of juvenile hormone III on Reticulitermes flavipes: changes in hemolymph protein composition and gene expression.

Termites express polyphenism during caste differentiation that is mostly undefined at the molecular level. Using the eastern subterranean termite, Reticulitermes flavipes Kollar, we wanted (1) to test juvenile hormone (JH) model assays for their ability to induce detectable molecular changes in worker termites and (2) to investigate hemolymph proteins and their corresponding genes during JH-induced soldier caste differentiation. Our results illustrate pronounced changes in two hemolymph proteins after JH treatment, as well as differences among several caste phenotypes. Significant increases in the expression of four genes encoding hemolymph proteins, including two vitellogenins and two hexamerins, were observed after JH exposure. These findings are the first to demonstrate such protein and gene expression changes during termite caste differentiation. These results also validate the utility of JH model assays for inducing detectable molecular changes in worker termites that have begun presoldier differentiation.

Animals↗

Hominins, sedges, and termites: new carbon isotope data from the Sterkfontein valley and Kruger National Park.

Stable carbon isotope analyses have shown that South African australopiths did not have exclusively frugivorous diets, but also consumed significant quantities of C4 foods such as grasses, sedges, or animals that ate these foods. Yet, these studies have had significant limitations. For example, hominin sample sizes were relatively small, leading some to question the veracity of the claim for australopith C4 consumption. In addition, it has been difficult to determine which C4 resources were actually utilized, which is at least partially due to a lack of stable isotope data on some purported australopith foods. Here we begin to address these lacunae by presenting carbon isotope data for 14 new hominin specimens, as well as for two potential C4 foods (termites and sedges). The new data confirm that non-C3 foods were heavily utilized by australopiths, making up about 40% and 35% of Australopithecus and Paranthropus diets respectively. Most termites in the savanna-woodland biome of the Kruger National Park, South Africa, have intermediate carbon isotope compositions indicating mixed C3/C4 diets. Only 28% of the sedges in Kruger were C4, and few if any had well-developed rhizomes and tubers that make some sedges attractive foods. We conclude that although termites and sedges might have contributed to the C4 signal in South African australopiths, other C4 foods were also important. Lastly, we suggest that the consumption of C4 foods is a fundamental hominin trait that, along with bipedalism, allowed australopiths to pioneer increasingly open and seasonal environments.

Animals↗

Characterization of termite lipophorin and its involvement in hydrocarbon transport.

The transport of lipids constitutes a vital function in insects and requires the plasma lipoprotein lipophorin. In all insects examined to date, cuticular hydrocarbons are also transported through the hemolymph by lipophorin, and in social insects they play important roles not only in water proofing the cuticle but also in nestmate recognition. High-density lipophorin (HDLp), isolated from Reticulitermes flavipes plasma by KBr gradient ultracentrifugation, contains 66.2% protein and 33.8% lipids; hydrocarbons constitute its major neutral lipid (20.4% of total lipids). Anti-lipophorin serum was generated in rabbit and its specific association with lipophorin, and not with any other plasma proteins, was verified with Western blotting. Immunoprecipitation also confirmed that this antibody specifically recognizes lipophorin, because all hemolymph hydrocarbons of the termites R. flavipes and R. lucifugus and the cockroach Supella longipalpa, which associate only with lipophorin, were recovered in the immunoprecipitated protein. Cross-reactivity of the antiserum with lipophorin from related species was investigated by double immunodiffusion with 10 termite species in the genera Reticulitermes, Coptotermes, Zootermopsis, and Kalotermes, and with five cockroach species. Involvement of lipophorin in hydrocarbon transport was shown by injecting HDLp antiserum into Zootermopsis nevadensis and then monitoring the de novo biosynthesis of hydrocarbons and their transport to the cuticular surface; the antiserum significantly disrupted hydrocarbon transport. ELISA revealed a gradual increase in the lipophorin titer in successively larger R. flavipes workers, and differences among castes in lipophorin titers were highest between nymphs and first instar larvae.

Animals↗

Phe-Gly-Leu-amide allatostatin in the termite Reticulitermes flavipes: content in brain and corpus allatum and effect on juvenile hormone synthesis.

In the subterranean termite Reticulitermes flavipes, allatostatins (ASTs) with the C-terminus Phe-Gly Leu-amide were localized by immunocytochemistry with antibody against a cockroach AST, Dippu AST-7. AST-immunoreactivity occurred in the corpus cardiacum and corpus allatum and in the lateral and medial neurosecretory cells of the brain that innervate these organs as well as in many other nerve cells of the brain. This was observed in workers, nymphs, soldiers and secondary reproductives. A radioimmunoassay, using anti-Dippu AST-11, demonstrated about 40 fmole equivalents of AST in brains of soldiers and secondary reproductives. The product of the corpora allata in this species was determined to be juvenile hormone III. Its synthesis by corpora allata of secondary reproductives, determined by in vitro radiochemical assay, was inhibited in a dose-dependent fashion by two cockroach allatostatins, Dippu AST-7 and Dippu AST-11. Thus, as in cockroaches and crickets, allatostatin-containing nerves innervate the corpora allata of this termite species and their production of juvenile hormone is inhibited by these neuropeptides.

Animals↗