Chromosomal organization of the 18S and 5S rRNAs and histone H3 genes in Scarabaeinae coleopterans: insights into the evolutionary dynamics of multigene families and heterochromatin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Chromosomal organization and evolutionary history of Mariner transposable elements in Scarabaeinae coleopterans

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Diet of the South American frog Leptodactylus ocellatus (Anura, Leptodactylidae) in Uruguay

The diet of Leptodactylus ocellatus (L., 1758) was studied in specimens collected at Maldonado Department (Uruguay), where monthly surveys were made between August 1998 and March 2000. A total of 143 frogs were collected, measured, sexed, and dissected, to assess stomach contents. The anurans were grouped into three age classes and four categories (considering sex and capture date). The trophic amplitude for each age class and sex-season category was quantified using Shannon-Weaver Index, and the trophic niche overlap between ages and categories by using the Pianka Index. Comparisons among treatments were made by G-tests. The most important prey items were coleopterans (IRI = 2547), and significant correlations were found between predator and prey sizes. The major differences on diet composition were found between extreme age classes (froglets and adults). Significant differences were also detected between sexes in the cold season but not in the warm season. Any of these changes in diet may be related with the availability of prey.

Diet and microhabitat use by two Hylodinae species (Anura, Cycloramphidae) living in sympatry and syntopy in a Brazilian Atlantic Rainforest area

We analyzed the diet and microhabitat use for two Hylodinae anurans (Cycloramphidae), Hylodes phyllodes Heyer & Cocroft, 1986 and Crossodactylus gaudichaudii Duméril & Bibron, 1841, living in sympatry at an Atlantic Rainforest area of Ilha Grande, in southeastern Brazil. The two species live syntopically at some rocky streams. The two species differed strongly in microhabitat use. Hylodes phyllodes occurred mainly on rocks, whereas C. gaudichaudii was observed mostly on the water. Regarding diet, coleopterans, hymenopterans (ants), and larvae were the most important prey item consumed by both species. Data suggest that microhabitat use appears to be an important parameter differentiating these frogs with respect to general resource utilization.

The first record of Litargus tetraspilotus (Coleoptera, Mycetophagidae) in Brazil, with biological notes and complementary description of the species

Litargus tetraspilotus LeConte, 1856 was collected feeding on Oidium sp. (Fungi, Ascomycota, Erysiphaceae) associated with fruit trees. This is the first time L. tetraspilotus is recorded in Brazil, totaling three species of Mycetophagidae for this country. This study aims to provide a complementary description of this species based on new characters and to present information on its life cycle under laboratory conditions and fluctuation in population in the field. During the period of inventories between July 2004 and August 2006, about every fifteen days, a total of 565 specimens of L. tetraspilotus were collected, with the highest abundance found on citrus plants, with values differing significantly between the two years. The population levels differed between the seasons; spring had the greatest abundance and autumn the least. There was a significant positive correlation of L. tetraspilotus abundance with rainfall and relative humidity. Mycetophagidae, as well as other mycophagous families of Brazilian coleopterans, are barely studied, warranting further future studies of their bioecology and systematics.

Succession and Abundance of Staphylinidae in Cattle Dung in Uberlândia, Brazil

Fimicolous Staphylinidae prey on rearing dipterous in cattle dung, acting as their natural controllers, including pests such as horn fly. To survey the abundance and succession of these coleopterans in cattle dung deposited in pasture, six experiments were conducted from March to October 1995 in Uberlândia, State of Minas Gerais, Brazil. Cattle dung pats were exposed at a pasture for 1 hr, 8 hr, 24 hr, 2, 3, 7 and 10 days and were than taken to laboratory separate from each other, for Staphylinidae extraction. A total of 156 dung pats were exposed at pasture, from which 6225 Staphylinidae were recovered. Representing at least 30 species. Staphylinidae sp.1 (29.6%), Philonthus flavolimbatus (22.2%), Heterothops sp.1 (16.6%), Oxytelus sp.2 (7.6%), Aleochara sp.2 (7.6%) and Criptobium sp.1 (4.4%) were the most abundant, representing 87.8% from the total. The increased frequency of the majority of these species along the dung exposition time at pasture, indicated tha, they would be preying on at all the immature stages of the dipterous, or eggs and first instar larvae of species that lay eggs on the dung after its second exposition day at the pasture

Biological aspects of Leucothyreus ambrosius Blanchard (Coleoptera, Melolonthidae, Rutelinae)

Biological aspects of Leucothyreus ambrosius Blanchard (Coleoptera, Melolonthidae, Rutelinae). Coleopterans of the family Melolonthidae comprise a large group of species that feed on different food sources, including plant roots, stems, and leaves, in addition to plant materials at different decomposition stages. Several species are found in the genus Leucothyreus, occurring in different regions of Brazil, including the various biomes in the country. Information on the biology of species of the genus Leucothyreus is scarce, therefore, we conducted studies on the biological aspects of Leucothyreus ambrosius Blanchard, 1850. The period of adult occurrence was determined with a light trap installed between a cropped and pasture area in the municipality of Aquidauana, Mato Grosso do Sul State, Brazil. Adults collected in the field were used to form insect pairs and the studies were initiated in the entomology laboratory as the adults began ovipositing. Adults were observed flying in the field from October to December. Eggs were obtained as pairs were formed and a colony was established, the embryonic period lasting 14.6 days on average. The larval period in the 1st instar lasted 21.6 days, in the 2nd instar 19.6 days, and in the 3rd instar, 85.6 days. The head capsule width was 1.48 mm in the 1st instar, 2.44 mm in the 2nd, and 3.83 mm in 3rd larval instar. The pupal stage had an average duration of 35.5 days. The egg to adult period lasted 173.3 days. Morphometric information for the larval and adult stages is presented in this study.

The effect of farming system and fertilisers on pests and natural enemies: A synthesis of current research

Organic agriculture is becoming widespread due to increased consumer demand and regulatory and political support. Organic agriculture can increase arthropod diversity but the response of pests and their natural enemies is variable. Fertiliser is an important component of agricultural systems and its effects on pests and natural enemies will influence agroecosystems. In this study, meta-analysis and vote-counting techniques were used to compare farming system (organic and conventional) and fertiliser effects on arthropod pests and their natural enemies. The meta-analyses indicated that pests generally benefitted from organic techniques, this is particularly evident when experiments were carried out on a smaller scale. Pest responses to organic and conventional fertiliser types were divergent, plant composts benefitted pest arthropods while the opposite was true for manures, this has implications for pest management. Most natural enemy groups responded positively to organic farming although this was not true for Coleopterans. Experimental scale had a prominent impact on natural enemy responses with farm scale studies showing particularly positive effects of organic agriculture on natural enemies. This suggests that it is large scale features of organic agriculture such as landscape heterogeneity that are beneficial to natural enemies. Natural enemy responses to organic fertilisers were positive indicating that field scale management practices including fertiliser can also be important in pest management.

Identificação de genes cry de Bacillus thuringiensis no controle de Sphenophorus levis, o bicudo da cana-de-açúcar

A bactéria B. thuringiensis caracteriza-se pela produção de proteínas tóxicas a representantes de diversas ordens de insetos, as quais são codificadas por genes cry. Este trabalho foi realizado com objetivo de selecionar isolados de B. thuringiensis, por meio da caracterização morfológica e molecular, identificando as diferentes subclasses dos genes cry3 e cry35 e determinar a patogenicidade contra Sphenophorus levis, uma das mais importantes pragas da cultura da cana-de-açúcar. Foram utilizados 1163 isolados de B. thuringiensis e com a observação em microscópio com contraste de fases foram confirmadas como pertencentes à espécie de B. thuringiensis. O material genético foi purificado pela matriz de troca iônica Instagene Matrix e submetido a PCR com iniciadores gerais cry3 e cry35 identificando-se 30 isolados contendo genes com potencial para o controle de coleópteros, os quais juntamente com as linhagens-padrão de B. thuringiensis var. tenebrionis, B. thuringiensis var. morrissone e B. thuringiensis var. tolworthi foram utilizados para a realização do bioensaio. Através de análise discriminante alocaram-se os isolados em quatro grupos quanto à toxicidade de B. thuringiensis. Os grupos ficaram assim definidos: um grupo que promovem até 10% de mortalidade contendo as testemunhas e duas linhagens;um grupo que causou 39% de mortalidade contendo três linhagens padrão e dez isolados; um grupo com 52% de mortalidade contendo treze isolados e um grupo com 70% de mortalidade contendo cinco isolados, os quais devem ser considerados promissores no controle biológico de S. levis.

Caracterização e seleção de isolados de Bacillus thuringiensis efetivos contra Sitophilus oryzae L., 1763

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização molecular e seleção de isolados de Bacillus thuringiensis com potencial inseticida para Sphenophorus levis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ecologia de dois grupos de sagüis-brancos, mico argentatus (Linnaeus 1771) em um fragmento florestal natural, Santarém-Pará

Este trabalho apresenta os resultados de um estudo de longo-prazo sobre ecologia e comportamento de dois grupos silvestres de Mico argentatus em um fragmento florestal natural, isolado por uma matriz de vegetação de savana, na região de Alter do Chão, Amazônia central. Os grupos, denominados Cm1 e Cm2 foram habituados e monitorados durante 12 meses em 2000 e quatro meses em 2001 (Cm1) e sete meses em 2000 e três meses em 2001 (Cm2). Análises de variação temporal envolveram três períodos sazonais distintos (Chuvosa-Início. Chuvosa-Final e Seca) em 2000, e a comparação longitudinal dos meses julho e setembro em 2000-2001. Análises das relações entre as variáveis ecológicas (abundância de principais recursos: frutos e insetos) e os padrões ecológicos e comportamentais entre os grupos foram estabelecidos. Os grupos apresentaram composições e tamanhos diferentes, que variaram entre 4-8 e 8-11 membros, em Cm1 e Cm2, respectivamente. Os dados quantitativos foram coletados utilizando-se a amostragem de varredura instantânea com amostras de um minuto de duração a cada intervalo de cinco minutos, durante todo o período de atividade diária do grupo. Os resultados mostraram que ambos os grupos gastam mais da metade do tempo em forrageio e alimentação. A dieta foi constituída de frutos, néctar, gomas e presa animal (invertebrados, principalmente ortópteros, formicídeos e coleópteros, além de lagartos e anfíbios). O tamanho total da área dos grupos Cm1 e Cm2 em 2000 foram respectivamente, 11,5 ha e 14,6 ha. As comparações sazonais mostraram uma similaridade entre grupos na alocação sistemática de mais tempo ao comportamento de forrageio e alimentação na medida em que os recursos não gomíferos se tornavam mais escassos. Já a análise longitudinal, além de apresentar uma redução de 30% na precipitação em 2001, mostrou uma mudança considerável na composição e distribuição espacial dos dois grupos. Assim, diferentes aspectos da comparação entre grupos pareceram refletir a influência destes diferentes fatores, com padrões opostos nos dois grupos. O aspecto geral mais importante parece ter sido o registro de padrões bem diferentes, além de similaridades importantes em grupos que ocupam quase o mesmo espaço, e a variabilidade das condições do hábitat entre as estações e entre os anos. As estratégias comportamentais identificadas pareceram ser reflexo, tanto da variabilidade e flexibilidade comportamental já conhecidas para os calitriquídeos, como de condições ecológicas impostas pela variabilidade na disponibilidade dos recursos num fragmento reduzido.

Diet Seasonality and Feeding Preferences of Brachycephalus pitanga (Anura: Brachycephalidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Recombinant expression, localization and in vitro inhibition of midgut cysteine peptidase (Sl-CathL) from sugarcane weevil, Sphenophorus levis

A cDNA coding for a digestive cathepsin L, denominated Sl-CathL, was isolated from a cDNA library of Sphenophorus levis larvae, representing the most abundant EST (10.49%) responsible for proteolysis in the midgut. The open reading frame of 972 bp encodes a preproenzyme similar to midgut cathepsin L-like enzymes in other coleopterans. Recombinant Sl-CathL was expressed in Pichia pastoris, with molecular mass of about 42 kDa. The recombinant protein was catalytically activated at low pH and the mature enzyme of 39 kDa displayed thermal instability and maximal activity at 37 degrees C and pH 6.0. Immunocytochemical analysis revealed Sl-CathL production in the midgut epithelium and secretion from vesicles containing the enzyme into the gut lumen, confirming an important role for this enzyme in the digestion of the insect larvae. The expression profile identified by RT-PCR through the biological cycle indicates that Sl-CathL is mainly produced in larval stages, with peak expression in 30-day-old larvae. At this stage, the enzyme is 1250-fold more expressed than in the pupal fase, in which the lowest expression level is detected. This enzyme is also produced in the adult stage, albeit in lesser abundance, assuming the presence of a different array of enzymes in the digestive system of adults. Tissue-specific analysis revealed that Sl-CathL mRNA synthesis occurs fundamentally in the larval midgut, thereby confirming its function as a digestive enzyme, as detected in immunolocalization assays. The catalytic efficiency of the purified recombinant enzyme was calculated using different substrates (Z-Leu-Arg-AMC, Z-Arg-Arg-AMC and Z-Phe-Arg-AMC) and rSl-CathL exhibited hydrolysis preference for Z-Leu-Arg-AMC (k(cat)/K-m = 37.53 mM S-1), which is similar to other insect cathepsin L-like enzymes. rSl-CathL activity inhibition assays were performed using four recombinant sugarcane cystatins. rSl-CathL was strongly inhibited by recombinant cystatin CaneCPI-4 (K-i = 0.196 nM), indicating that this protease is a potential target for pest control. (C) 2011 Elsevier Ltd. All rights reserved.

Evaluation of the pathogenicity of multiple isolates of Beauveria bassiana (Hypocreales: Clavicipitaceae) on Rhynchophorus ferrugineus (Coleoptera: Dryophthoridae) for the assessment of a solid formulation under simulated field conditions

A solid state formulation of Beauveria bassiana (Balsamo) Vuillemin has been developed for biological control of the Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier, 1790). Two kinds of bioassays (dry conidia and dipping) using 10 isolates from several coleopterans in Mediterranean environments, identified 2 RPW derived isolates (193 and 203) as most pathogenic to RPW larvae and adults (zero survival within first 4–5 d for dry conidia, and 14 and 23 d for dipping bioassays). Isolate 203 (5.1 × 108 ± 1.9 × 108 conidia g-1) was formulated with fragmented date seed into solid granules and tested in palms infested with RPW under semi-field conditions in Feb, Apr/May and Jun of both 2007 and 2008. Beauveria bassiana significantly reduced RPW adult survival with respect to controls in May 2007 and in the Apr/Jun 2008 experiments. Total RPW adult mortality was achieved within 30 days for all B. bassiana treatments, and was associated with increasing numbers of insects with signs of mycosis in 2008 experiments. Beauveria bassiana formulation reduced RPW multiplication in artificially infested palms compared to controls, and a positive correlation between numbers of larvae and time post-infestation was recorded. The suppression of RPW adult populations by B. bassiana persisted for at least 3 months under semi-field conditions. The Beauveria bassiana solid formulation, which induces great adult mortality and persistence in the field, could be applied as a preventive as well as a curative treatment for the integrated management of RPW.