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Sporulation of Metarhizium anisopliae and Beauveria bassiana on Coptotermes formosanus and in vitro.

The sporulation of 22 total isolates of Metarhizium anisopliae and Beauveria bassiana was quantified on cadavers of the Formosan subterranean termite, Coptotermes formosanus. Conidial production increased significantly over 11 days post-death. Effects of isolates of M. anisopliae and B. bassiana on in vivo sporulation were significant. Although the overall effects of fungal species on in vivo sporulation were not significant, the interactions between fungal species and certain times post-death were significant, indicating different sporulation patterns between the two fungal species. B. bassiana isolates could be categorized into a group with high total sporulation (day 11) and low quick sporulation (on days 2 and 3), while M. anisopliae isolates fell into another group with high quick sporulation and low total sporulation. This could give M. anisopliae an advantage over B. bassiana in termite microbial control due to termite defensive social behaviors. Conidial production was significantly higher in vitro than in vivo. In vitro and in vivo sporulation differed by as much as 89x and 232x among the selected isolates of B. bassiana and M. anisopliae, respectively. Correlation between in vivo and in vitro conidial production was positive and significant. This may allow preliminary in vitro screening of a large number of isolates for high in vivo sporulation.

Animals↗

Effects of virulence, sporulation, and temperature on Metarhizium anisopliae and Beauveria bassiana laboratory transmission in Coptotermes formosanus.

Sporulation characteristics and virulence of Metarhizium anisopliae and Beauveria bassiana were examined in relation to laboratory transmission in Coptotermes formosanus. Fungal isolates significantly affected disease prevalence in termite populations. Sporulation of M. anisopliae played a more important role than virulence in producing epizootics within small groups of termites, but this was not the case for B. bassiana. Isolates characterized by quick sporulation (day 2 after death) did not exhibit better transmission in termites than those with high total sporulation (day 11 after death) in either fungal species. An isolate of M. anisopliae ranking highly in all three categories (virulence, quick sporulation, and total sporulation) produced better epizootics than an isolate that was inferior in all three characteristics. High temperatures (35 degrees C) significantly reduced fungal germination rates, leading to significant reduction of epizootics. M. anisopliae was better than B. bassiana in producing epizootics at 27 degrees C. Thus, fungal characteristics other than virulence should be considered for the seasonal colonization approach to termite microbial control.

Animals↗

Composition of the essential oil of Lepidium meyenii (Walp).

The essential oil profile of maca (Lepidium meyenii) obtained from Lima, Peru, was examined. Steam distillates of the aerial parts of L. meyenii were continuously extracted with pentane and the pentane extracts analyzed by GC/MS. Retention indices and mass spectral data were used to identify 53 oil components. Phenyl acetonitrile (85.9%), benzaldehyde (3.1%), and 3-methoxyphenylacetonitrile (2.1%) were the major components of the steam distilled oil. The oil of L. meyenii was tested for phytotoxic, cyanobactericidal, and antitermite activity. The oil was selectively toxic towards the cyanobacterium Oscillatoria perornata compared to the green alga Selenastrum capricornutum, with complete growth inhibition at 100 microg/ml. Mortality of the Formosan subterranean termite, Coptotermes formosanus, was numerically, but not significantly, higher when held on filter paper treated with maca oil. At 1% (w/w), maca oil also appeared to act as a feeding deterrent to termites. Several minor components of the essential oil of maca including 3-methoxyphenylacetonitrile and benzylthiocyanate were significantly active against the Formosan termite. This is the first report on the essential oil composition of L. meyenii.

Animals↗

Development of environmentally-benign wood preservatives based on the combination of organic biocides with antioxidants and metal chelators.

Wood extractives can be envisaged to protect heartwood by at least three different mechanisms, i.e. fungicide, free radical scavengers/antioxidants and as metal chelators. In short-term laboratory decay tests using two different wood species and decay fungi, the combination of different organic fungicides with various antioxidants and/or metal chelators gave enhanced activity as compared to the organic biocide alone, with the best results usually obtained with all three compounds. Outdoor ground-contact stakes treated with a biocide and antioxidant combination and exposed for 30 months also gave enhanced protection against both decay fungi and termites. It was concluded that the combination of an organic biocide with metal chelating and/or antioxidant additives gives enhanced protection to wood against fungi as compared to the biocide alone and, consequently, it may be possible to develop environmentally-benign wood preservative systems based on this idea.

Animals↗

Intersexual variations in the venom of the Brazilian 'armed' spider Phoneutria nigriventer (Keyserling, 1891).

Venom from male and female specimens of the medically most important Brazilian Ctenidae spider Phoneutria nigriventer (Keyserling, 1891) has been compared. Males showed a slightly higher venom yield (386 microg) than equal sized females (296 microg), while adult females showed nearly a three times higher venom yield (1079 microg). High-pressure liquid-chromatography analyses revealed differences in the venom composition between males and females. A single peak in HPLC (peak 11) was only detected in venom from females carrying egg-sacs, and sodium dodecyl sulfate-gel electrophoresis showed a series of high molecular weight proteins only in the male venom pool. The median lethal dose (LD(50)) in mice for female venom was 0.63 microg kg(-1) (95% confidence interval [0.54; 0.71] and 0.61 [0.56; 0.73] microg kg(-1)) for females with egg-sacs, when compared to the male venom which showed a LD(50) of 1.57 [1.46; 1.88] microg kg(-1). The venom of both sexes was also tested in insects using a termite bioassay with doses of 2, 3, 4, and 5 microg per termite. No effect was detected for the lowest dose of female venom, whereas all the other venom doses from both sexes caused a decreased paralysis time and death of the termites. Comparing the venom of both sexes, it was observed that female venom provoked a faster reaction than male venom. The results indicate that males and females of P.nigriventer have differing venom composition which lead to different effects in biological assays.

Animals↗

Paralizing activity of the Parawixia bistriata crude venom in termites: a new bioassay.

Spider venoms have high specificity to neuronal elements. Therefore, the use of venom has been important in the characterisation of mammal and insect nervous systems. The evaluation of insect paralysis has been an important tool for distinguishing the biological effects of venom. In this study we describe the paralysing effect of a spider crude venom (Parawixia bistriata) in termites, utilising a new bioassay. The crude venom of P. bistriata caused an irreversible and dose-dependent paralysis in the animals in the following doses: 2.10(-5) U; 2.10(-4) U; 2.10(-3) U; 2.10(-2) U and 0.12 U (1 U = 1 gland). This bioassay will allow for easy and direct evaluation of biological effects from different venoms and purified fractions.

Animals↗

Potential antitermite compounds from Juniperus procera extracts.

Thin layer chromatography (TLC) analysis revealed that destructive distillation of Juniperus procera tree gave ten major components, whereas Croton megalocarpus tree yielded five components. This was confirmed by gas chromatography (GC). The components were isolated by column chromatography and analysed using infrared, ultra-violet, visible and mass spectroscopy (MS) techniques. The whole extract was about 30.3% of the starting material (sawdust) and consisted of 77.5% water and 22.5% oily reddish-brown layer. The extracts had alcoholic and phenolic compounds together with acids. Cedrol, a tertiary tricyclic alcohol, was found to be in the greatest proportion in the oily layer. IR spectra with a peak beyond 3000 cm(-1), UV-VIS absorption maxima at 230 nm and mass spectra with m/e 204 suggested the presence of cedrene in the extract.

Animals↗

In situ measurements of metabolite fluxes: microinjection of radiotracers into insect guts and other small compartments.

In microbial ecology, it is of great interest to determine metabolic activities under in situ conditions, i.e., without disturbing the structure of the community and the spatial arrangement of individual populations by experimental manipulation. Microinjection of radiotracers and subsequent analysis using the isotope dilution technique has proven to be a powerful method to measure metabolic fluxes in small biological systems, e.g., the intestinal tract of termites. The large variety of commercially available radiolabeled substrates and the identification and quantitation of radiolabeled products by chromatographic methods allow for investigation of the complete metabolic network in a given system.

Animals↗

Metazoan cellulase genes from termites: intron/exon structures and sites of expression.

Endogenous endo-beta-1,4-glucanase (EGase, EC 3.2.1.4) cDNAs were cloned from representatives of the termite families Termitidae and Rhinotermitidae. These EGases are all composed of 448 amino acids and belong to glycosyl hydrolase family 9 (GHF9), sharing high levels of identity (40-52%) with selected bacterial, mycetozoan and plant EGases. Like most plant EGases, they consist of a single catalytic domain, lacking the ancillary domains found in most microbial cellulases. Using a PCR-based strategy, the entire sequence of the coding region of NtEG, a gene putatively encoding an EGase from Nasutitermes takasagoensis (Termitidae), was determined. NtEG consists of 10 exons interrupted by 9 introns and contains typical eukaryotic promoter elements. Genomic fragments of EGase genes from Reticulitermes speratus (Rhinotermitidae) were also sequenced. In situ hybridization of N. takasagoensis guts with an antisense NtEG RNA probe demonstrated that expression occurs in the midgut, which contrasts to EGase expression being detected only in the salivary glands of R. speratus. NtEG, when expressed in Escherichia coli, was shown to have in vitro activity against carboxymethylcellulose.

Animals↗

Ethidium bromide: a fast fluorescent staining procedure for the detection of symbiotic partnership of flagellates and prokaryotes.

The hindgut of 'lower' termites harbors a dense population of flagellates and bacteria. The flagellates possess ecto- and endosymbiotic prokaryotes. Most of them are hardly visible in the phase contrast microscope. Staining with the DNA-intercalating agent ethidium bromide visualizes the nuclei of the flagellates as well as the ecto- and endosymbiotic bacteria as red objects. Furthermore, it is possible to distinguish between endosymbiotic methanogens and other bacteria. Following UV excitation, the blue-green autofluorescence of the methanogenic bacteria eclipses the red fluorescence light of the intercalated ethidium bromide. The dye facilitates the observation of symbiotic bacteria and helps identify the number, shape, localization, and dividing status of the nuclei. Thus, it is a powerful tool for the examination of microorganisms in complex habitats, which are rich in strongly autofluorescent material, like wood.

Animals↗

Isolation and identification of a diuretic hormone from Zootermopsis nevadensis.

A diuretic hormone (DH) was isolated from extracts of heads of Zootermopsis nevadensis, a dampwood termite. The peptide has 46 residues, M(r) = 5,328.2 Da, with the sequence TGAVPSLSIVNPLDVLRQRLLLEIARRRMRQSQDQIQANREMLQTI-NH(2,) showing it to be a CRF-related DH. This peptide increases cyclic AMP production in Malpighian tubules of Manduca sexta. We detected another factor in the head extracts which behaved as a more basic peptide on ion exchange chromatography. The latter factor also stimulated cyclic AMP production in the bioassay, but two large scale attempts to isolate this peptide were unsuccessful. We believe the second peptide is acid labile.

Amino Acid Sequence↗

Evaluation of primers and PCR conditions for the analysis of 16S rRNA genes from a natural environment.

We investigated biases occurring in the polymerase chain reaction (PCR) amplification of 16S rRNA genes from an environmental sample, by comparing the clone libraries that we had previously prepared from the gut homogenate of the termite Reticulitermes speratus. We detected a significant increase in the expected number of phylotypes by lowering the annealing temperature, and a significant decrease in the proportion of clones belonging to the predominant group by raising the number of PCR cycles. We also found that the Bacteria-universal primer, 63F, introduced a seriously biased amplification caused by primer mismatches, in contrast to a previous report. These results, together with suggestions from previous studies using simplified model samples, will help us to recognize the limitations of PCR-based analysis.

Animals↗

Estimating occupational exposure to the pyrethroid termiticide bifenthrin by measuring metabolites in urine.

The control of subterranean termites in Australia is predominantly through the application of chemical barriers in the soil beneath and surrounding buildings. The chemicals used to repel or kill termites are the organophosphorus insecticide, chlorpyrifos, and the synthetic pyrethroid, bifenthrin. These are applied through surface sprays and subfloor injection by licensed pest control operators. To determine the exposure of these personnel to these pesticides it is most usual to measure airborne concentrations or dermal deposition rates. However, to support information obtained from these methods it is often appropriate to determine the amount of the chemicals absorbed, using biological monitoring techniques including measurement of the chemicals or their metabolites in urine. While there are effective techniques for the monitoring of chlorpyrifos exposure by measuring either the alkyl phosphate or trichloropyridinol metabolites, there have been no published reports of suitable methods to measure bifenthrin metabolites in urine. This paper describes an extraction and HPLC-UV method used to simultaneously measure the urinary excretion of 2-methyl-3-phenylbenzoic acid (MPA), a metabolite of bifenthrin, and 3-phenoxybenzoic acid (PBA), a metabolite of several other common pyrethroid insecticides, with a detection limit for each of 2.5 ng/ml. The paper also describes the pilot application of this method to a study of South Australian pest control operators handling bifenthrin. MPA ranged from 1.8 to 31.9 microg/g creatinine and PBA from 1.3 to 30.0 microg/g in the urine of pest control workers. MPA was detected in urine of control workers without bifenthrin exposure only at low levels (1.0-1.4 microg/g creatinine), but PBA was found in both at higher levels (1.2-61.1 microg/g creatinine).

Adult↗

Acute otalgia: a case report of mature termite in the middle ear.

Acute otalgia during childhood is one of the most common complaints in general ENT practice. It may occur as a result of acute otitis externa, otitis media or a foreign body. Animate foreign body in the ear canal or in the middle ear usually results in otalgia and hearing loss. We present a rare case of a living mature termite in the middle ear of a 9-year-old girl complaining of intermittent attacks of otalgia associated with a loud cracking sound in the left ear. In this case, a small perforation of the tympanic membrane without a history of previous trauma, infection, or evidence of a foreign body in the external ear raises a suspicion. A careful, time-consumed microscopic examination could show the living creature in the middle ear. Immediate immobilization and removal of the living foreign body are imperative. In conclusion, tympanic membrane perforation and intermittent severe otalgia without history of otitis may lead to a suspicion of any insects in the middle ear.

Acute Disease↗

Demographic mechanisms for the evolution of long life in social insects.

One of the most remarkable life history traits of many species of eusocial insects is the extraordinary longevity of their queens, with the recorded life span of some individuals exceeding 30 years. Surprisingly, little research has been conducted on either the proximate or ultimate questions concerning what factors favor the evolution of the long life spans in social insects. The broad objective of this paper is to address four sets of questions: (1) What are the similarities and differences in the longevity patterns among species in the four main groups of social insects - bees, wasps, ants and termites? (2) What is the evolutionary association of longevity and sociality in insects? (3) Are there biodemographic principles that can be derived from studies on longevity in social insects? and (4) What are the general implications for longevity in vertebrates, including humans?

Animals↗

[Properties of the structures produced by ecology engineers at the soil surface of a Colombian savanna].

Fourteen types of physical structures produced by ecosystem engineers were sampled at the surface of a savanna soil of Colombia. Invertebrates implicated in the creation of these structures were identified. Some physical (aggregate size and stability, bulk density) and chemical (C, N, P contents, pH, etc) properties of structures were assessed. Three large groups of structures were identified: (i) earthworm casts characterised by a high bulk density (1.3-1.4 g.cm-3), constituted of aggregates (7-10 mm), high in organic C (3-4%) and assimilable nutrients; (ii) termite mounds with low bulk density (0.6-0.9 g.cm-3), constituted of aggregates (8-9 mm), high in organic C (3.5-10%) and assimilable nutrients; and (iii) slightly compact (0.4-0.7 g.cm-3) and granular (aggregate size < 1.5 mm) termite superficial channels and ant mounds low in organic C (less than 1.5%) and assimilable nutrients. These results underline the large diversity of the biogenic structures produced at the surface of the studied soil. They suggest the feasibility of a functional classification of engineer organisms that would take into account simultaneously the different functional attributes reflected by these structures.

Animals↗

Evaluation of acute sensory--motor effects and test sensitivity using termiticide workers exposed to chlorpyrifos.

Sensory and motor testing was performed on a group of termiticide workers primarily using chlorpyrifos-containing products to evaluate both the acute effects from current exposure and sensitivity of the measures to detect effects. The study group comprised 106 applicators and 52 nonexposed participants. Current exposure was measured by urinary concentrations of 3,5,6-trichloro-2-pyridinol (TCP) collected the morning of testing. The mean TCP value for the 106 applicators was 200 microg/g creatinine. Participants received 4--5 h of testing and were evaluated using a sensory--motor test battery recommended by a National Institute for Occupational Safety and Health (NIOSH)-sponsored advisory panel to be appropriate for testing effects from pesticide exposures. Measurements testing olfactory dysfunction, visual acuity, contrast sensitivity, color vision, vibrotactile sensitivity, tremor, manual dexterity, eye--hand coordination, and postural stability were analyzed. Study results indicated limited acute effects from exposure to chlorpyrifos using urinary TCP as a measure of current exposure. The effects occurred primarily on measures of postural sway in the eyes closed and soft-surface conditions, which suggests a possible subclinical effect involving the proprioceptive and vestibular systems. Several other tests of motor and sensory functions did not show any evidence of acute exposure effects, although statistically significant effects of urinary TCP on the Lanthony color vision test scores and one contrast sensitivity test score were found. The visual measures, however, were not significant when a step-down Bonferroni correction was applied. Information also is presented on the sensitivity of the measures to detect effects in an occupationally exposed population using standard error of the parameter estimates.

Animals↗

Devescovinid trichomonad with axostyle-based rotary motor ("Rubberneckia"): taxonomic assignment as Caduceia versatilis sp. nov.

An amitochondriate trichomonad cell of the family Devescovinidae (Class Parabasalia), helped demonstrate the fluid model of lipoprotein cell membranes. This wood-ingesting symbiont in the hindgut of the dry wood-eating termite Cryptotermes cavifrons is informally known to cell biologists as "Rubberneckia". As the microtubular axo-style complex generates force causing clockwise movement of the entire anterior portion of the cell at the shear zone the protist displays "head" rotation. Studies by phase contrast and videomicroscopy of live cells, of whole mounts by scanning, and thin sections by transmission electron microscopy extend the observations of Tamm and Tamm [24-26] and Tamm [19-23]. Habitat, cell shape, size, nuclear features, parabasal apparatus and other morphological details permit the assignment of "Rubberneckia" to Kirby's cosmopolitan genus Caduceia. This large-sized devescovinid has distinctive parabasal gyres, an axostylar rotary, motor, and regularly-associated nonflagellated, fusiform and flagellated rod epibiotic surface bacteria. In addition to regularly aligned epibionts intranuclear and endocytoplasmic bacteria are abundant and hydrogenosomes are Present. "Rubberneckia" is compared here to the other seven species of Caduceia. Since it is clearly sufficiently distinctive to warrant new species status, we named it C. versatilis.

Animals↗