Aspidosperma (Apocynaceae) plant cytotoxicity and activity towards malaria parasites. Part I: Aspidosperma nitidum (Benth) used as a remedy to treat fever and malaria in the Amazon

Infusions of Aspidosperma nitidum (Apocynaceae) wood bark are used to treat fever and malaria in the Amazon Region. Several species of this family are known to possess indole alkaloids and other classes of secondary metabolites, whereas terpenoids, an inositol and the indole alkaloids harmane-3 acid and braznitidumine have been described in A. nitidum . In the present study, extracts from the wood bark, leaves and branches of this species were prepared for assays against malaria parasites and cytotoxicity testing using human hepatoma and normal monkey kidney cells. The wood bark extracts were active against Plasmodium falciparum and showed a low cytotoxicity in vitro, whereas the leaf and branch extracts and the pure alkaloid braznitidumine were inactive. A crude methanol extract was subjected to acid-base fractionation aimed at obtaining alkaloid-rich fractions, which were active at low concentrations against P. falciparum and in mice infected with and sensitive Plasmodium berghei parasites. Our data validate the antimalarial usefulness of A. nitidum wood bark, a remedy that can most likely help to control malaria. However, the molecules responsible for this antimalarial activity have not yet been identified. Considering their high selectivity index, the alkaloid-rich fractions from the plant bark might be useful in the development of new antimalarials.

Conservation and developmental expression of ubiquitin isopeptidases in Schistosoma mansoni

Several genes related to the ubiquitin (Ub)-proteasome pathway, including those coding for proteasome subunits and conjugation enzymes, are differentially expressed during the Schistosoma mansoni life cycle. Although deubiquitinating enzymes have been reported to be negative regulators of protein ubiquitination and shown to play an important role in Ub-dependent processes, little is known about their role in S. mansoni . In this study, we analysed the Ub carboxyl-terminal hydrolase (UCHs) proteins found in the database of the parasite’s genome. An in silicoana- lysis (GeneDB and MEROPS) identified three different UCH family members in the genome, Sm UCH-L3, Sm UCH-L5 and SmBAP-1 and a phylogenetic analysis confirmed the evolutionary conservation of the proteins. We performed quantitative reverse transcription-polymerase chain reaction and observed a differential expression profile for all of the investigated transcripts between the cercariae and adult worm stages. These results were corroborated by low rates of Z-Arg-Leu-Arg-Gly-Gly-AMC hydrolysis in a crude extract obtained from cercariae in parallel with high Ub conjugate levels in the same extracts. We suggest that the accumulation of ubiquitinated proteins in the cercaria and early schistosomulum stages is related to a decrease in 26S proteasome activity. Taken together, our data suggest that UCH family members contribute to regulating the activity of the Ub-proteasome system during the life cycle of this parasite.

Aspidosperma (Apocynaceae) plant cytotoxicity and activity towards malaria parasites. Part II: experimental studies withAspidosperma ramiflorum in vivo and in vitro

Several species of Aspidosperma plants are used to treat diseases in the tropics, including Aspidosperma ramiflorum, which acts against leishmaniasis, an activity that is experimentally confirmed. The species, known as guatambu-yellow, yellowperoba, coffee-peroba andmatiambu, grows in the Atlantic Forest of Brazil in the South to the Southeast regions. Through a guided biofractionation of A. ramiflorum extracts, the plant activity against Plasmodium falciparum was evaluated in vitro for toxicity towards human hepatoma G2 cells, normal monkey kidney cells and nonimmortalised human monocytes isolated from peripheral blood. Six of the seven extracts tested were active at low doses (half-maximal drug inhibitory concentration < 3.8 µg/mL); the aqueous extract was inactive. Overall, the plant extracts and the purified compounds displayed low toxicity in vitro. A nonsoluble extract fraction and one purified alkaloid isositsirikine (compound 5) displayed high selectivity indexes (SI) (= 56 and 113, respectively), whereas compounds 2 and 3 were toxic (SI < 10). The structure, activity and low toxicity of isositsirikine in vitro are described here for the first time in A. ramiflorum, but only the neutral and precipitate plant fractions were tested for activity, which caused up to 53% parasitaemia inhibition of Plasmodium bergheiin mice with blood-induced malaria. This plant species is likely to be useful in the further development of an antimalarial drug, but its pharmacological evaluation is still required.

Impact of a gall midge Parkiamyia paraensis (Diptera, Cecidomyiidae) on the Amazonian plant Parkia pendula (Fabaceae)

We observed the occurrence of large numbers of galls induced by Parkiamyia paraensis (Diptera, Cecidomyiidae) on the leaflets of Parkia pendula (Fabaceae) in northern Para, Brazil. We addressed two questions in this study: i) what is the proportion of attacked plants in the field, and nursery conditions?; and ii) what is the impact of galls on the host plant? An average of 86% of the plants were galled in the field. Galled P. pendula were distinct from healthy individuals due to their prostrated architecture and death of terminal shoots. Approximately 50% of the total available leaves and 35% leaflets were attacked by P. paraensis on saplings under nursery conditions. Each one-year old plant supported an average of 1,300 galls, and an average of 60g allocated to galled tissue. Otherwise, attacked individuals were taller and heavier than healthy plants. Attacked plants weighed five times more than healthy plants. When the weight of the galls was removed, the total weight (aerial part without galls) of attacked plants was drastically reduced, indicating that most of the biomass of attacked plants was due to the attack by P. paraensis galls. Although the data indicate a paradox, as young plants attacked by the galling herbivore appear to develop more vigorously than unattacked plants, we suggest that P. paraensis negatively affect P. pendula development.

Relationships between host plant architecture and gall abundance and survival

The plant architecture hypothesis predicts that variation in host plant architecture influences insect herbivore community structure, dynamics and performance. In this study we evaluated the effects of Macairea radula (Melastomataceae) architecture on the abundance of galls induced by a moth (Lepidoptera: Gelechiidae). Plant architecture and gall abundance were directly recorded on 58 arbitrarily chosen M. radula host plants in the rainy season of 2006 in an area of Cerrado vegetation, southeastern Brazil. Plant height, dry biomass, number of branches, number of shoots and leaf abundance were used as predicting variables of gall abundance and larval survival. Gall abundance correlated positively with host plant biomass and branch number. Otherwise, no correlation (p > 0.05) was found between gall abundance with shoot number or with the number of leaves/plant. From a total of 124 galls analyzed, 67.7% survived, 14.5% were attacked by parasitoids, while 17.7% died due to unknown causes. Larvae that survived or were parasitized were not influenced by architectural complexity of the host plant. Our results partially corroborate the plant architecture hypothesis, but since parasitism was not related to plant architecture it is argued that bottom-up effects may be more important than top-down effects in controlling the population dynamics of the galling lepidopteran. Because galling insects often decrease plant fitness, the potential of galling insects in selecting for less architectural complex plants is discussed.

Rediscovery of the putatively extinct ant species Simopelta minima (Brandão) (Hymenoptera, Formicidae), with a discussion on rarity and conservation status of ant species

Simopelta minima (Brandão, 1989) was originally described based on four workers collected in soil samples from a small cocoa plantation in Ilhéus, state of Bahia, northeastern Brazil. In the subsequent years after the description, this cocoa plantation was eliminated and the species was then considered extinct by the Brazilian environmental institutions. The recent rediscovery of S. minima workers in subterranean pitfall trap samples from Viçosa, state of Minas Gerais, southeastern Brazil, over 1.000 km distant from type locality, suggests that the rarity and vulnerability status of some ant species may be explained by insufficient sampling of adequate microhabitats, in time and space.

Are gall midge species (Diptera, Cecidomyiidae) host-plant specialists?

Despite the speciose fauna of gall-inducing insects in the Neotropical region, little is known about their taxonomy. On the other hand, gall morphotypes associated with host species have been extensively used as a surrogate of the inducer species worldwide. This study reviewed the described gall midges and their galls to test the generalization on the use of gall morphotypes as surrogates of gall midge species in the Brazilian fauna. We compiled taxonomic and biological data for 196 gall midge species recorded on 128 host plant species. Ninety two percent of those species were monophagous, inducing galls on a single host plant species, whereas only 5.6% species were oligophagous, inducing galls on more than one congeneric host plant species. Only four species induced galls on more than one host plant genus. We conclude that gall morphotypes associated with information on the host plant species and attacked organs are reliable surrogates of the gall-inducing species.

Gall midge attack intensity and host-plant response in a Neotropical coastal ecosystem

The gall inducer Clusiamyia nitida Maia, 1996 (Diptera, Cecidomyiidae) often infests the shrub Clusia lanceolata (Camb.) (Clusiaceae) in the Neotropical vegetation of restinga of Rio de Janeiro State, Brazil. Leaves of Clusia lanceolata host up to 20 spheroid galls and show variation in their shape. We aimed to evaluate the effect of gall's intensity on leaves of Clusia lanceolata, and the extension of gall's impact on adjacent non-galled leaves. We analyzed the effect of the number of galls on leaf area, biomass, specific area and leaf appearance from 509 leaves of 14 individual plants. The results showed that differences of individual plants, pairs of leaves, and gall presence were responsible for more then 90% of variation on infested leaves. Variation on parasitic intensity level created differences in leaf response. Under moderate gall attack characterized by scattered galls on a leaf, the increase of the number of galls caused an increase of leaf biomass and area, and a decrease of specific area. The specific area was smaller also under high attack intensity, characterized by coalescent galls on a leaf. In those cases of extremely high parasitic intensity, galled leaves became deformed and the surface area was severely reduced. Leaf deformation due to gall attack led to early leaf abscission, indicated by the 90% of deformed leaves found in the youngest leaf pair of the branch. There was insufficient evidence that the impact of galls on leaf morpho-physiological parameters extended beyond the attacked leaves, because ungalled leaves did not change significantly when their opposite leaf had been galled.

Effects of sex, host-plant deprivation and presence of conspecific immatures on the cannibalistic behavior of wild Ascia monuste orseis (Godart) (Lepidoptera, Pieridae)

Effects of sex, host-plant deprivation and presence of conspecific immatures on the cannibalistic behavior of wild Ascia monuste orseis (Godart) (Lepidoptera, Pieridae). The specialist cabbage caterpillar Ascia monuste orseis (Lepidoptera, Pieridae) feeds on plants of the Brassicaceae family, but may eventually ingest conspecific eggs and larvae during the larval stage. The present study examines feeding behavior of 4th and 5th instar cabbage caterpillars in relation to sex, host-plant deprivation and presence of conspecifics. We recorded number of egg ingested per larvae, developmental indices and duration of feeding, exploratory and resting behavior. Kale deprived caterpillars presented high rates of cannibalism, development delay and decreased fecundity. Cannibalism rates were not influenced by the sex of the larvae. In general, the presence of conspecific eggs did not interfere with the frequency and duration of the categorical behavioral events. We conclude that food availability is a strong factor influencing the extent to which A. monuste orseis caterpillars cannibalize.

Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus) (Coleoptera, Curculionidae)

Toxic effects of essential plant oils in adult Sitophilus oryzae (Linnaeus) (Coleoptera, Curculionidae). Stored grains are subject to losses in quality nutritional value and in sanitation from the time they are stored to the time they are consumed. Botanical insecticides may offer an alternative solution for pest control. The objective was to test the insecticidal properties of the essential oils of Cymbopogon citratus (leaf), Zingiber officinale (root) and Mentha sp. (leaf). The efficacy of these oils was tested to control the rice weevil, S. oryzae, using hydrodistillation. Chemical analysis of the essential oils was carried out by gas chromatography. Major components of C. citratus were geranial (48%) and neral (31%), of Z. officinale were α-zingibereno (13%), geranial (16%), neral (10%) and α-farneseno (5%) and of Mentha sp. was menthol (92%). Bioassays were carried out by fumigation and topical application. In topical application assays, the essential oil of C. citratus had greater toxicity (LC50 0.027 µL mL-1) and shorter exposure time than the oils of the other two plants. After 24 h and 48 h, 70% and 100% mortality of S. oryzae occurred, respectively. In fumigation assays, essential oil of Z. officinale had a lower LC50 (1.18 µL cm-2) and 70% mortality after 24 h exposure. Therefore, we recommend the use of essential oils of C. citratus and Z. officinale to control the rice weevil S. oryzae.

Strepsicrates smithiana Walsingham (Lepidoptera, Tortricidae): first record from Chile and a newly documented host plant

Strepsicrates smithiana Walsingham (Lepidoptera, Tortricidae): first record from Chile and a newly documented host plant. Strepsicrates smithiana Walsingham, 1892 (Lepidoptera, Tortricidae) is recorded for the first time from Chile. Male and female adults were reared from leaf-tying larvae collected on Myrica pavonis (Myricaceae), which is a new host plant record for S. smithiana.

Population growth and within-plant distribution of the striped mealybug Ferrisia virgata (Cockerell) (Hemiptera, Pseudococcidae) on cotton

Population growth and within-plant distribution of the striped mealybug Ferrisia virgata (Cockerell) (Hemiptera, Pseudococcidae) on cotton. The striped mealybug, Ferrisia virgata (Cockerell) (Hemiptera, Pseudococcidae), is a widely distributed and polyphagous pest species, which naturally occurs on cotton plants in Brazil. This study evaluated the establishment and population growth as well as the within-plant distribution of F. virgata on four cotton cultivars: CNPA 7H (white fibers), BRS Verde, BRS Safira, and BRS Rubi (colored fibers). The experiment was conducted in a complete randomized design with four treatments (cultivars) and 18 replications of each. Thus, cotton plants of each cultivar were infested with 100 newly hatched nymphs of F. virgata. The number of adult female mealybugs and the total number of mealybugs per plant were quantified, respectively, at 25 and 50 days after infestation. The developmental and pre-reproductive periods were also determined. Furthermore, we verified the distribution of F. virgata on the plant parts at 25 and 50 days after infestation. Ferrisia virgata showed similar growth of 412-fold in the four cotton cultivars studied. Also, the nymphs were spread on infested leaves; the secondgeneration nymphs were spread and established in all plant parts. Our results characterize F. virgata as having much potential as an important cotton pest in Brazil.

First host plant records for Iridopsis hausmanni Vargas (Lepidoptera, Geometridae) in the coastal valleys of northern Chile

First host plant records for Iridopsis hausmanni Vargas (Lepidoptera, Geometridae) in the coastal valleys of northern Chile. The trees Haplorhus peruviana Engl. and Schinus molle L. (Anacardiaceae) are mentioned as the first host plant records for the little known native moth Iridopsis hausmanni Vargas, 2007 (Lepidoptera, Geometridae, Ennominae) in the coastal valleys of the northern Chilean Atacama Desert. This is also the first record of Anacardiaceae as host plant for a Neotropical species of Iridopsis Warren, 1894.

An unusual food plant for Cydia pomonella (Linnaeus) (Lepidoptera, Tortricidae) in Mexico

An unusual food plant for Cydia pomonella (Linnaeus) (Lepidoptera, Tortricidae) in Mexico. Larvae of Cydia pomonella (Linnaeus, 1758) were discovered on floral cones of Magnolia schiedeana (Schltdl, 1864) near the natural reserve of La Martinica, Veracruz, México. Magnolia represents an unusual host for this moth species, which is known throughout the world as the "codling moth", a serious pest of fruits of Rosaceae, especially apples. The larvae were identified using taxonomic keys, and identification was corroborated using molecular markers. Further sampling resulted in no additional larvae, hence, the observation was probably that of an ovipositional error by the female, and M. schiedeana is not at risk of attack by this important moth pest.

Egg laying site selection by a host plant specialist leaf miner moth at two intra-plant levels in the northern Chilean Atacama Desert

Egg laying site selection by a host plant specialist leaf miner moth at two intra-plant levels in the northern Chilean Atacama Desert. The spatial distribution of the immature stages of the leaf miner Angelabella tecomae Vargas & Parra, 2005 was determined at two intra-plant levels (shoot and leaflet) on the shrub Tecoma fulva fulva (Cav.) D. Don (Bignoniaceae) in the Azapa valley, northern Chilean Atacama Desert. An aggregated spatial pattern was detected for all the immature stages along the shoot, with an age dependent relative position: eggs and first instar larvae were clumped at apex; second, third and fourth instar larvae were mostly found at intermediate positions; meanwhile the spinning larva and pupa were clumped at basis. This pattern suggests that the females select new, actively growing leaflets for egg laying. At the leaflet level, the immature stages were found more frequently at underside. Furthermore, survivorship was higher for larvae from underside mines. All these results highlight the importance of an accurate selection of egg laying site in the life history of this highly specialized leaf miner. By contrast, eventual wrong choices in the egg laying site selection may be associated with diminished larval survivorship. The importance of the continuous availability of new plant tissue in this highly human modified arid environment is discussed in relation with the observed patterns.