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Measurement of oxygen partial pressure, its control during hypoxia and hyperoxia, and its effect upon light emission in a bioluminescent elaterid larva.

This study investigates the respiratory physiology of bioluminescent larvae of Pyrearinus termitilluminans in relation to their tolerance to hypoxia and hyperoxia and to the supply of oxygen for bioluminescence. The partial pressure of oxygen (P(O2)) was measured within the bioluminescent prothorax by in vivo electron paramagnetic resonance (EPR) oximetry following acclimation of larvae to hypoxic, normoxic and hyperoxic (normobaric) atmospheres and during periods of bioluminescence (during normoxia). The P(O2) in the prothorax during exposure to an external P(O2) of 15.2, 160 and 760 mmHg was 10.3+/-2.6, 134+/-0.9 and 725+/-73 mmHg respectively (mean +/- s.d., N=5; 1 mmHg=0.1333 kPa). Oxygen supply to the larvae via gas exchange through the spiracles, measured by determining the rate of water loss, was also studied in the above atmospheres and was found not to be dependent upon P(O2). The data indicated that there is little to no active control of extracellular tissue P(O2) within the prothorax of these larvae. The reduction in prothorax P(O2) observed during either attack-response-provoked bioluminescence or sustained feeding-related bioluminescence in a normoxic atmosphere was variable, but fell within the range 10-25 mmHg. The effect of hypoxic atmospheres on bioluminescence was measured to estimate the intracellular P(O2) within the photocytes of the prothorax. Above a threshold value of 50-80 mmHg, bioluminescence was unaffected by P(O2). Below this threshold, an approximately linear relationship between P(O2) and bioluminescence was observed. Taken together with the extracellular P(O2) measurements, this suggests that control of P(O2) within the photocyte may occur. This work establishes that EPR oximetry is a valuable technique for long-term measurement of tissue P(O2) in insects and can provide valuable insights into their respiratory physiology. It also raises questions regarding the hypothesis that bioluminescence can have a significant antioxidative effect by reduction of prothorax P(O2 )through oxygen consumption.

Animals↗

Three new sesquiterpenes from the black heartwood of Cryptomeria japonica.

Three new sesquiterpenes, (4S)-2,6,10-bisaboratrien-4-ol-1-one (1), 1,8-epoxy-1(6),2,4,7,10-bisaborapenta-en-4-ol (2), and 1-methoxy-4-cadinene (3) have been isolated from the black heartwood of Cryptomeria japonica. Compounds 1 and 2 were designated sugikurojinols A and B, respectively, and the structures of compounds 1-3 were established by extensive NMR experiments. Compounds 1 and 2 were also examined for antibacterial activity against Staphylococcus aureus and Escherichia coli, and for termiticidal activity against Coptotermes formosanus SHIRAKI.

Animals↗

Insect antifeedants, pterocarpans and pterocarpol, in heartwood of Pterocarpus macrocarpus Kruz.

The insect antifeedant activities of pterocarpans and a sesquiterpene alcohol from the dichloromethane extract of Pterocarpus macrocarpus Kruz. (Leguminosae) were evaluated against the common cutworm, Spodoptera litura F. (Noctuidae), and the subterranean termite, Reticulitermes speratus (Kolbe)(Rhinotermitidae). Three pterocarpans, (-)-homopterocarpin (1), (-)-pterocarpin (2), and (-)-hydroxyhomopterocarpin (3) and the sesquiterpene alcohol, (+)-pterocarpol (5), were isolated from the dichloromethane extract of the heartwood of P. macrocarpus under guidance by a biological assay. Among these natural products, the most active insect antifeedant against both S. litura and R. speratus was 1. On the other hand, sesquiterpene alcohol 5 showed less insect antifeedant activity than the other pterocarpans against both insect species. While its methylated derivative, (-)-methoxyhomopterocarpin (4), showed high biological activity, 3 showed less insect antifeedant activity in this study. Interestingly, racemic 1 did not show insect antifeedant activity against S. litura. However, all of the test pterocarpans and isoflavones showed antifeedant activity against the test termites. Additionally, since these compounds were major constituents of P. macrocarpus, these antifeedant phenolics may act as chemical defense factors in this tree. In Thailand, lumber made from this tree is used to make furniture and in building construction due to its resistance to termite attack.

Alcohols↗

Distribution and properties of endo-beta-1,4-glucanase from a lower termite, Coptotermes formosanus (Shiraki).

A multi-enzyme distribution of endo-beta-1,4-glucanase activity was found in the digestive system of a worker caste of the lower termite Coptotermes formosanus (Shiraki) by zymogram analysis. Its distribution analysis demonstrated that about 80% of this activity was localized in salivary glands from where only one component (EG-E) was secreted into the digestive tract. EG-E was isolated by a combination of chromatographic and electrophoretic techniques. Its molecular mass, optimal pH and temperature, isoelectric point, and Km were 48 kDa, 6.0, 50 degrees C, 4.2, and 3.8 (mg/ml on carboxymethylcellulose), respectively. EG-E hydrolyzed cellooligosaccharides with a degree of polymerization of 4 and larger, and had low activity on crystalline cellulose. Main reaction products from low molecular weight cellulose were cellobiose and cellotriose. The N-terminal amino acid sequence of EG-E has similarity with fungal endo-beta-1,4-glucanases and cellobiohydrolases of the glycosyl hydrolase family 7 rather than the other insect endo-beta-1,4-glucanases of family 9.

Amino Acid Sequence↗

Molecular phylogeny of symbiotic basidiomycetes of fungus-growing termites in Thailand and their relationship with the host.

Termitomyces-related symbiotic basidiomycetes in the nests of fungus-growing termites (Macrotermitinae) of several genera in Thailand were cultivated and analyzed phylogenetically based on the DNA sequence of nuclear ribosomal RNA genes. The relationships of the symbiotic fungi with host termites and their locality were apparently complex, supporting intricate mechanisms for the termites to acquire the symbionts.

Animals↗

Diverse bacteria related to the bacteroides subgroup of the CFB phylum within the gut symbiotic communities of various termites.

Phylogenetically diverse clones of the partial 16S rDNA (ca. 850 bp) of bacteria belonging to the bacteroides subgroup of the cytophaga-flavobacter-bacteroides phylum were collected from the symbiotic microbial communities in the guts of six termite species without cultivation. Combined with the sequences reported previously, a total of thirty phylotypes of the subgroup were identified and classified into five phylogenetic clusters. One that was comprised of the phylotypes from a single termite species was related to the genus Rikenella. Two were clustered each with some cultured strains, genera of which have not been clearly defined yet. The remaining two clusters had no culturable representatives, suggesting the presence of yet-uncultivated genera within the termite guts. From these sequence data, we designed a specific primer for the bacteroides subgroup, which was successful in the terminal-restriction fragment length polymorphism analysis to detect the phylotypes of the subgroup in the termite gut.

Animals↗

Isolation of actinomycetes from termites' guts.

Actinomycetes could be isolated efficiently on a defatted wood powder medium from the guts of various species of termites. The actinomycete flora in the termites' guts depended largely on the area in which the termites naturally occur. In termites from the same area, the actinomycete flora changed depending on the taxonomic difference between termite species. Some actinomycetes isolated from termites' guts grew satisfactorily on lignin-related media, and the others grew on cellulose-related media.

Actinobacteria↗

Isolation and characterization of aromatics-degrading microorganisms from the gut of the lower termite Coptotermes formosanus.

We isolated aromatics-degrading bacteria from the gut of a lower termite, Coptotermes formosanus, using a mineral salt medium containing various aromatic compounds as the sole carbon source. Two species, Burkholderia sp. strain VE22 and Citrobacter sp. strain VA53, were isolated by aerobic enrichment culture with veratraldehyde and vanillin, respectively. Strain VA53 could also grow and metabolize vanillin anaerobically.

Aerobiosis↗

Molecular phylogenetic diversity of the bacterial community in the gut of the termite Coptotermes formosanus.

The phylogenetic diversity of the bacterial community in the gut of the termite Coptotermes formosanus was investigated using a 16S rRNA gene clone library constructed by PCR. After screening by restriction fragment length polymorphism (RFLP) analysis, 49 out of 261 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. Many of the clones (94%) were derived from Bacteroidales, Spirochaetes, and low G + C content gram-positive bacteria consisting of Clostridiales, Mycoplasmatales, Bacillales, and Lactobacillales. In addition, a few clones derived from Actinobacteria, Proteobacteria, Planctomycetes, Verrucomicrobia, and the candidate phylum "Synergistes" were also found. The most frequently identified RFLP type, BCf1-03, was assigned to the order Bacteroideales, and it constituted about 70% of the analyzed clones. The phylogenetic analysis revealed that the representative clones found in this study tended to form some clusters with the sequences cloned from the termite gut in several other studies, suggesting the existence of termite-specific bacterial lineages.

Animals↗

Heterologous overexpression of a mutant termite cellulase gene in Escherichia coli by DNA shuffling of four orthologous parental cDNAs.

Among cellulase genes, those of animals are known for their difficulty in overexpression. We constructed a chimeric library by family shuffling of endo-beta-1,4-glucanase genes from four different termite species (Reticulitermes speratus, Nasutitermes takasagoensis, Coptotermes formosanus, and Coptotermes acinaciformis) sharing 78.5-96% homology in amino acid sequence. The constructed library was screened by Congo red plate assay combined with 96-well micro-enzyme assay, and clones showing enhanced CMCase activities were obtained. The mutated genes were overexpressed in Escherichia coli intracellularly as an active form. The endo-beta-1,4-glucanase (CMCase) activity in soluble fractions of E. coli harboring the mutant genes was 20-30 fold higher than that of wild-type genes. The mutant enzyme showed high activity against CMC and properties similar to those of the native enzymes.

Amino Acid Sequence↗

Phylogenetic and morphological diversity of Bacteroidales members associated with the gut wall of termites.

The microbial community adherent directly or indirectly to the gut wall of termites is distinct from that of the other habitats in the gut. The bacterial 16S rRNA genes were identified from the fractionated gut walls of two termite species, Hodotermopsis sjoestedti and Neotermes koshunensis, and compared with those previously identified from Reticulitermes speratus. Surprisingly, the bacterial constituents were almost entirely different among the termites at the phylotype level (the criterion of the phylotype was >97% nucleotide identity). Bacteria in the order Bacteroidales, which were commonly abundant symbionts on gut walls, were phylogenetically analyzed. They were dispersed in a number of clusters formed by phylotypes from the guts of various termites. In situ hybridization with probes specific for some phylotypes and a phylogenetic cluster detected the cells of several Bacteroidales members with a significant variety of cell morphology in the gut wall fractions, which reflects the phylogenetic diversity of this order.

Animals↗

Morphological adaptation of the skull for various behaviors in the tree shrews.

Skull size and shape were examined among 14 species of the tree shrews (Tupaia montana, T. picta, T. splendidula, T. mulleri, T. longipes, T. glis, T. javanica, T. minor, T. gracilis, T. dorsalis, T. tana, Dendrogale melanura, D. murina, and Ptilocercus lowii). The bones of face were rostro-caudally longer in T. tana and T. dorsalis, contrasting with T. minor and T. gracilis, D. melanura, D. murina and P. lowii which have smaller facial length ratios. The arbo-terrestrial species (T. longipes and T. glis) were similar to terrestrial species in length ratios of bones of face unlike the other arbo-terrestrial species (T. montana, T. picta, T. splendidula, and T. mulleri). We propose that T. longipes and T. glis have adapted to foraging for termites and ants as have T. tana and T. dorsalis. Additionally small body size in T. javanica may be the result of being isolated in Java. We separated the species into 5 groups from the measurment values of skulls: 1) Terrestrial species; T. tana and T. dorsalis, 2) Arboreal species; T. minor and T. gracilis, 3) Arbo-terrestrial species group 1: T. montana, T. splendidula, T. picta and T. mulleri, and T. javanica, 4) Arbo-terrestrial species group 2: T. glis and T. longipes, 5) Arboreal species of Dendrogale and Ptilocercus. Principal component analysis separated species into 8 clusters as follows: 1) T. tana, 2) T. dorsalis, 3) T. montana, T. splendidula, T. picta and T. mulleri, 4) T. glis and T. longipes, 5) T. javanica, 6) T. minor and T. gracilis, 7) D. melanura and D. murina, and 8) P. lowii. We suggest that these clusters correspond to behavioral strategies and peculiarities observed in foraging, feeding and locomotion in each species.

Adaptation, Physiological↗

Protection of Vochysia elliptica (Vochysiaceae) by a nectar-thieving ant.

Vochysia elliptica (Vochysiaceae) is a shrubby plant, which does not have EFNs. Camponotus ants thieve nectar, and can decrease plant fitness by making flowers less attractive to pollinators. However, ants remove herbivores, wHich can be beneficial. Results show that plants from which ants were excluded had lower rates of termite (simulated herbivore) removal than did plants visited by ants. Plants accessible to ants showed higher rates of termite removal in the base of leaves and in the inflorescence, than in the tip of leaves. This occurs because ants must pass through the principal axis to reach the inflorescence. Conclusive results of this cost/benefit analysis of the Camponotus sp. presence for V. elliptica can be obtained, with experimental manipulations.

Animals↗

Ant and termite mound coinhabitants in the wetlands of Santo Antonio da Patrulha, Rio Grande do Sul, Brazil.

This paper reports on ant and termite species inhabiting the mounds (murundus) found in three wetland sites in Santo Antonio da Patrulha. Ants and termites were found in 100% of the mounds of two sites and in 20% of those in the third site. Colonies of Camponotus fastigatus were found inhabiting all the mounds, while colonies of Brachymyrmex sp., Linepithema sp., Pheidole sp., and/or Solenopsis sp. were collected in less than 30% of the mounds. In the mounds of the three sites, colonies of Anoplotermes sp. and/or Aparatermes sp. termites were found together with the ant colonies. Another cohabiting termite species, Cortaritermes sp., was found only in the mounds of one site. The results suggest that C. fastigatus is the species building the mounds, with the other species, whether ants or termites, being the inquilines.

Animals↗

Geophone detection of subterranean termite and ant activity.

A geophone system was used to monitor activity of subterranean termites and ants in a desert environment with low vibration noise. Examples of geophone signals were recorded from a colony of Rhytidoponera taurus (Forel), a colony of Camponotus denticulatus Kirby, and a termite colony (undetermined Drepanotermes sp.) under attack by ants from a nearby C. denticulatus colony. The geophone recordings were compared with signals recorded from accelerometers in a citrus grove containing Solenopsis invicta Buren workers. Because of their small size, all of these insects produce relatively weak sounds. Several different types of insect-generated sounds were identified in the geophone recordings, including high-frequency ticks produced by R. taurus and C. denticulatus, and patterned bursts of head bangs produced by Drepanotermes. The S. invicta produced bursts of ticks with three different stridulation frequencies, possibly produced by three different-sized workers. Overall, both systems performed well in enabling identification of high-frequency or patterned pulses. The geophone was more sensitive than the accelerometer to low-frequency signals, but low-frequency insect sound pulses are more difficult to distinguish from background noises than high-frequency pulses. The low cost of multiple-geophone systems may facilitate development of future applications for wide-area subterranean insect monitoring in quiet environments.

Acoustics↗

Penetration of the eastern subterranean termite into soil treated at various thicknesses and concentrations of Dursban TC and Premise 75.

The ability of eastern subterranean termites, Reticulitermes flavipes (Kollar), to penetrate various concentrations and treatment thicknesses ranging from 1.0 to 50.0 mm of Dursban TC and Premise 75 was evaluated in a laboratory bioassay. Termites penetrated only a few millimeters into 500 ppm Dursban TC-treated soil at all thicknesses. As concentration decreased, termite penetration into Dursban TC-treated soil increased with termites generally penetrating completely through 5.0- and 0.5-ppm treatments. Termites penetrated at least 30% into 10-, 25-, and 50-mm thicknesses of 100 ppm Premise 75. Termites generally completely penetrated all concentrations < or = 100 ppm and thicknesses < or = 5 mm. At 7 d, termite mortality was 100% at all treatment thicknesses of Dursban TC at 500 and 50 ppm. At 5.0 and 0.5 ppm Dursban TC, termite mortality ranged from approximately 45 to 98% across all treatment thicknesses. At 100- and 10-ppm concentrations of Premise 75-treated soil, termites suffered > or = 75% mortality. Premise 75 at 1.0 and 0.1 ppm generally killed < or = 50% of the assayed termites at all treatment thicknesses.

Animals↗

A computerized system for remote monitoring of subterranean termites near structures.

Sensors comprising wooden stakes painted with conductive circuits of silver particle emulsion were inserted in Sentricon stations in soil near structures. Sensors were wired to a datalogger that was programmed to test for circuit breakage every 2 h and store the data in its memory. A host computer was programmed to access the datalogger through telephone communication lines for data download every 4 d. The computerized monitoring system was tested in three remote sites, and site visits were conducted monthly for 6 mo to examine system accuracy in detecting termite activity. The mean monthly accuracy for the system to correctly report the presence (true positive) or absence of termites (true negative) in the stations was 85%, but the accuracy at 6 mo after system installation ranged from 41 to 79%. Mean sensor longevity, defined as the time for a sensor circuit to break in the absence of termites, was approximately 4.4 mo.

Animals↗