Electroantennographic responses of Heterotermes tenuis (Isoptera: Rhinotermitidae) to synthetic (3Z,6Z,8E)-Dodecatrien-1-ol

The subterranean termite Heterotermes tenuis (Isoptera: Rhinotermitidae) is a pest of great importance for the Brazilian economy as it causes serious damages in commercial reforestations of Eucalyptus spp. and sugarcane cultures. (3Z,6Z,8E)-Dodecatrien-1-ol has been identified as a pheromone of some species of subterranean termites. The objective of this research was to synthesize (3Z,6Z,8E)-dodecatrien-1-ol and determine by electroantennographic (EAG) bioassays the selectivity and sensitivity of H. tenius antennae to its whole worker extract and to the synthetic triene alcohol.

Physiological and cellular responses in two populations of the mussel Perna perna collected at different sites from the Coast of São Paulo, Brazil

The physiological conditions of mussels from Ubatuba and Santos and also of organisms transplanted from Ubatuba to Santos were studied by using different techniques. Assays for lysosomal stability were conducted on the haemolymph. Heart rate activity was monitored for 6h. The embryonic development of larvae obtained from the collected mussels was analysed. For all the compared groups of mussels, no significant differences were observed for the cardiac activity monitoring and the embryonic bioassays. The mean Neutral Red (NR) retention time was similar for the animals from Santos and Ubatuba, whereas the organisms transplanted to Santos showed a reduction in the retention time of the dye, indicating damage in the lysosomal membranes. These differences were possibly due to environmental factors, but further investigations are required to confirm this hypothesis.

Oral toxicity of chemical substances found in Dimorphandra mollis (Caesalpiniaceae) against honeybees (Apis mellifera) (Hymenoptera: Apidae)

Dimorphandra mollis is a characteristic plant from Brazilian saP vanna-like vegetation. The pollen of this species could be toxic to bees and the objective of the present investigation was to study the toxicity of methanolic extracts obtained from the flowers, peduncles and stem bark of D. mollis to Apis mellifera workers. For the study, the extracts were incorporated into the diet of the bees for later evaluation of mortality rates. The substances isolated: neoisoastilbin, catechin, astilbin and tannins were tested on adult workers and only catechin did not cause toxic effects. The data obtained in the toxicity bioassays were analyzed statistically by Log Rank test and all methanolic extracts showed significant (p<0.0001) toxic effects. Astilbin is also the major component of pollen grains, and is probably responsible for honeybee mortality during blooming periods.

Astilbin toxicity to leaf-cutting ant Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The toxicity of astilbin, isolated from Dimorphandra mollis, was tested in laboratory ingestion bioassays for the leaf-cutting ant Atta sexdens rubropilosa Forel. Worker ants that were fed an artificial diet daily to which astilbin was added had a higher mortality rate than the controls. The substance astilbin was incorporated into the diet with and without the utilization of a solvent. Results for both methodologies utilized were similar and indicate that astilbin reduces the median survival of ants. All concentrations tested showed toxic effects against ant workers, and the statistical comparison of survivorship rates from control and treated groups was significantly different, confirming the insecticidal properties of the substance astilbin.

Toxicity of Cipadessa fruticosa to the leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and their symbiotic fungus

Crude extracts from fruits, branches and leaves of Cipadessa fruticosa were assayed on Atta sexdens rubropilosa workers and their symbiotic fungus Leucoagaricus gongylophorus. The results showed that the majority of the extract tested reduced significantly (p < 0.05) the survival of the workers comparable to the neem oil. In addition, the dichloromethane extract from fruits and the hexane extract from branches of C. fruticosa inhibited in 80% the development of the symbiotic fungus. The hexane and dichloromethane extracts from fruits of C. fruticosa were fractionated and their fractions were subjected to new bioassays. Several fractions showed relevant activity on ants, but none of them inhibited significantly the fungal growth. The possibility of controlling leaf-cutting ants in the future using C. fruticosa is discussed.

Brosimum gaudichaudii Trecul (Moraceae): Da planta ao medicamento

The chemical, biological, pharmacological and toxicological study of plants used in medical treatments by different societies around the world has been intensifying over the years, primarily in the search for new herbal, homeopathic or even allopathic medicines. Motivated by this interest, we are investigating some Brazilian plant species, among them a tree, Brosimum gaudichaudii Trecul (Moraceae), found in Brazil's savannah-like prairies and scrub-forest (cerrado) and mesophytic forest. Several extracts have been made and separated into fractions and some compounds have been isolated - principally coumarins - and their respective structures analyzed by spectrophotometric and other methods. By collaborating with researchers in other fields, we have been able to carry out various bioassays and even basic clinical tests, namely: determination of anti-helminthic, anti-microbial, photosensitizing, anticancer and mutagenic activities, toxicological and pathogenic studies, with the aim of discovering a genuinely Brazilian herbal or homeopathic medicine.

Toxicity of straight-chain fatty acids to leaf-cutting ants Atta sexdens rubropilosa (Hymenoptera: Formicidae) and the symbiotic fungus Leucoagaricus gongylophorus

This work determined toxicity and attractiveness of straight-chain fatty acids (C 5 to C 12) to Atta sexdens rubropilosa (Forel) workers. The effect to the symbiotic fungus, Leucoagaricus gongylophorus (Singer) Möller, was also tested with the fatty acids C 6 to C 12. A strong mortality of leaf-cutting ants that were fed with an artificial diet containing fatty acids C to C at concentrations above 1.0 mg.ml -1 was observed. Rice flakes impregnated with solutions of these fatty acids were repellent to leaf-cutting ants. Contact experiments showed that treatments with C 6 and C 7 at concentration of 100 mg.ml -1 significantly reduced the survival rate of leaf-cutting ants. The fatty acids C 8 to C 11 were toxic to leaf-cutting ants when topically tested at concentration of 200 mg.ml -1. In relation to the fungus' bioassays, the fatty acids C 6 to C 12 at concentration of 0.1 mg.ml -1 inhibited 100% of the fungal development. Although when the concentration was reduced by half no inhibition effects were observed. The results showed that straight-chain fatty acids have desirable properties for controlling leaf-cutting ants since they directly interfere with both organisms of the symbiotic relationship. The potential of fatty acids as well as ways to control leaf-cutting ants with these compounds are discussed in this article.

Seletividade de alguns agrotóxicos em uso na citricultura ao ácaro predador Neoseiulus californicus (McGregor) (Acari: Phytoseiidae)

This work was carried out to determine the residual toxicity of the main pesticides used in citrus, on Neoseiulus californicus (McGregor) under laboratory conditions. For this, it was used the residual contact bioassay. Citrus leaves of the variety Pêra were sprayed in a Potter tower. The residual action was evaluated at 2 hours and 1; 3; 5; 7; 10; 14 e 21 days after treatment. Ten adult females of N. californicus were transferred to each ring, together with an enough amount of Tetranychus urticae nymphs and adults to feed the predator. Mortality evaluations were performed at 72 hours after treatment. Bioassays indicate that the pesticides: acrinathrin, deltamethrin, dinocap, sulphur, fenpropathrin, fenbutatin oxide and propargite were innocuous to N. californicus. On the other hand, abamectin, azocyclotin e cyhexatin caused mortalities of 29.8; 32.0 e 34.1%, respectively, two hours after transference. Dicofol, pyridaben and chlorfenapyr were extremely toxic to the predator mite, causing 100% of mortality two hours after the application. This way, to this population of N. californicus, the releases can be performed safely 3 days after the application, except for cyhexatin (5 days), dicofol and pyridaben (14 days) and chlorfenapyr (21 days), without risk of significant adult mortality rates due to the application of pesticides.

Interspecific interactions between Coptotermes gestroi and Heterotermes tenuis (Isoptera: Rhinotermitidae) under laboratory conditions

Coptotermes gestroi and Heterotermes tenuis are termite pests. Both species have cryptic habits, and are known as subterranean termites. The aim of this study was to verify if food availability and alterations in the proportion of soldiers interfere in the interactions between these termites, since the introduced species C. gestroi occupies the same niche as the native species H. tenuis. Different bioassays were conducted in order to evaluate these interspecific interactions. The results showed that C. gestroi presented advantages in the occupation of territory in all bioassays and suggest a higher efficiency of this species in the search for alternative food sources, especially when the number of soldiers was higher than that of H. tenuis. Additionally, it is possible to conclude that food availability and the different proportion of soldiers involved in the interspecific confrontations interfere in the occupation of territory and in the agonistic behavior between these two species of termites.

Toxoplasma gondii in semen of experimentally infected swine

Eight reproductive boars were divided into three groups and inoculated with Toxoplasma gondii [GI (n=3) 1.5×104 oocysts strain P; GII (n=3) 1.0×106 tachyzoites strain RH; and GIII (n=2) non-inoculated control]. Clinical, hematological, parasitemia and serological tests and studies of the parasite in the semen through bioassay and PCR, and in reproductive organs (Bioassay and immunohistochemical analyses) were conducted to evaluate the toxoplasmic infection. Blood and semen were collected on day -2, -1, 1, 3, 5, 7, 9, 11, 14 and weekly up to 84 days post-inoculation (DPI). No clinical or hematimetric alteration was observed in the boars. Parasitemia was detected in one boar inoculated with oocysts at the 7th DPI and in another boar infected with tachyzoites (GII) at the 3rd and 49 th DPI. Serological tests revealed antibodies against T. gondii in animals inoculated with oocysts or tachyzoites at the 7th DPI with dilutions of 1:256 and 1:64, which reached peaks of 1:4096 at day 11 and 9, respectively. The bioassays revealed the presence of the parasite in semen samples of a boar inoculated with oocysts (GI) at 3, 49 and 56 DPI and from two boars infected with tachyzoites (GII), one animal at 5 and two animals at 49 days DPI. Mice inoculated with semen from the control group (GIII) remained serologically negative. PCR analysis showed T. gondii DNA in the semen of Boar 1 and Boar 3 inoculated with tachyzoites and oocysts, respectively. The immunohistochemical tests showed T. gondii in the reproductive organs of Boar 1 and Boar 2, inoculated with tachyzoites and oocysts, respectively. These findings suggest the possible occurrence of venereal transmission of T. gondii in swine.

Laboratory assessment of two active ingredients for control of coptotemes gestroi (Isoptera: Rhinotermitidae)

The subterranean termite Coptotermes gestroi Wasmann (Isoptera: Rhinotermitidae) is a pest of major economic importance in urban environments of southeastern Brazil. Compared to using pesticides for termite control, termite bait products target termites more specifically and also address environmental contamination issues. In this point of view, we performed two different bioassays in order to evaluate the efficacy of boric acid and fipronil against different populations of C. gestroi. The results showed that concentrations between 2000 and 3000 ppm of boric acid caused approximately 100 percent mortality in termites. Concentrations between 0.01 and 0.0001 ppm of fipronil resulted in 100% termite mortality after 2 wk exposure. The data displayed a fast mortality of termites contaminated with fipronil, even with small concentrations, and therefore it is not a suitable product to be used in baits against C. gestroi. The present study showed a delayed toxicity of boric acid against the subterranean termite C. gestroi which suggests a need for further field tests.

Toxicological and histopathological effects of boric acid on atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The current study compared the toxicity of different concentrations of boric acid in adult workers of Atta sexdens rubropilosa Forel (Hymenoptera: Formicidae), with toxicological bioassays, and examining the dose-dependent and time-dependent histopathological changes, of the midgut, Malpighian tubules, and postpharyngeal glands. Our results revealed the importance of conducting toxicological bioassays combined with morphological analyses of the organs of ants chronically exposed to insecticides used in commercial ant baits. In vitro bioassays showed that boric acid significantly decreases the survivorship of workers regardless of concentration, whereas the morphological data suggested progressive dose-dependent and time-dependent changes in the organs examined, which were evident in the midgut. The midgut is the first organ to be affected, followed by the postpharyngeal gland and Malpighian tubules. This sequence is in agreement with the absorption pathway of this chemical compound in the midgut, its transference to the hemolymph, possibly reaching the postpharyngeal glands, and excretion by the Malpighian tubules. These progressive changes might be due to the cumulative and delayed effect of boric acid. Our findings provide important information for the understanding of the action of boric acid in ant baits in direct and indirect target organs. © 2010 Entomological Society of America.

Expression of a new Bacillus thuringiensis Cry1Ia gene in Escherichia coli with strong activity against cotton pests

The control of cotton pests may be accomplished using Bacillus thuringiensis Cry proteins. For this purpose, the objective of this work was to evaluate the insecticidal activity of a new Cry1Ia protein against neonatal larvae of Spodoptera frugiperda and Anthonomus grandis. The complete cry1Ia gene, previously obtained by PCR with oligonucleotide primers based on the sequenced gene, was cloned into the vector pET28a(+), introduced into Escherichia coli BL21(DE3) and expressed by induction with IPTG. The expression of the Cry1Ia protein was confirmed with molecular weight of approximately 81 kDa. The results demonstrated the efficiency of the bacterial system for the expression of B. thuringiensis Cry1Ia protein, which was subsequently used in quantitative bioassays against S. frugiperda and A. grandis larvae, resulting in an extremely toxic protein for both species. This characteristic is exceptionally important for obtaining transgenic cotton plants resistant to these pests.

The role of multiple partners in a digestive mutualism with a protocarnivorous plant

Background and aimsThe protocarnivorous plant Paepalanthus bromelioides (Eriocaulaceae) is similar to bromeliads in that this plant has a rosette-like structure that allows rainwater to accumulate in leaf axils (i.e. phytotelmata). Although the rosettes of P. bromelioides are commonly inhabited by predators (e.g. spiders), their roots are wrapped by a cylindrical termite mound that grows beneath the rosette. In this study it is predicted that these plants can derive nutrients from recycling processes carried out by termites and from predation events that take place inside the rosette. It is also predicted that bacteria living in phytotelmata can accelerate nutrient cycling derived from predators.MethodsThe predictions were tested by surveying plants and animals, and also by performing field experiments in rocky fields from Serra do Cipó, Brazil, using natural abundance and enriched isotopes of 15N. Laboratory bioassays were also conducted to test proteolytic activities of bacteria from P. bromelioides rosettes.Key ResultsAnalyses of 15N in natural nitrogen abundances showed that the isotopic signature of P. bromelioides is similar to that of carnivorous plants and higher than that of non-carnivorous plants in the study area. Linear mixing models showed that predatory activities on the rosettes (i.e. spider faeces and prey carcass) resulted in overall nitrogen contributions of 26·5 % (a top-down flux). Although nitrogen flux was not detected from termites to plants via decomposition of labelled cardboard, the data on 15N in natural nitrogen abundance indicated that 67 % of nitrogen from P. bromelioides is derived from termites (a bottom-up flux). Bacteria did not affect nutrient cycling or nitrogen uptake from prey carcasses and spider faeces.ConclusionsThe results suggest that P. bromelioides derive nitrogen from associated predators and termites, despite differences in nitrogen cycling velocities, which seem to have been higher in nitrogen derived from predators (leaves) than from termites (roots). This is the first study that demonstrates partitioning effects from multiple partners in a digestion-based mutualism. Despite most of the nitrogen being absorbed through their roots (via termites), P. bromelioides has all the attributes necessary to be considered as a carnivorous plant in the context of digestive mutualism. © 2012 The Author. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.

Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera)

The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65. kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana. © 2012 Elsevier Inc.