Tsetse thrombin inhibitor: Bloodmeal-induced expression of an anticoagulant in salivary glands and gut tissue of Glossina morsitans morsitans

The tsetse thrombin inhibitor, a potent and specific low molecular mass (3,530 Da) anticoagulant peptide, was purified previously from salivary gland extracts of Glossina morsitans morsitans (Diptera: Glossinidae). A 303-bp coding sequence corresponding to the inhibitor has now been isolated from a tsetse salivary gland cDNA library by using degenerate oligonucleotide probes. The full-length cDNA contains a 26-bp untranslated segment at its 5′ end, followed by a 63-bp sequence corresponding to a putative secretory signal peptide. A 96-bp segment codes for the mature tsetse thrombin inhibitor, whose predicted molecular weight matches that of the purified native protein. Based on its lack of homology to any previously described family of molecules, the tsetse thrombin inhibitor appears to represent a unique class of naturally occurring protease inhibitors. Recombinant tsetse thrombin inhibitor expressed in Escherichia coli and the chemically synthesized peptide are both substantially less active than the purified native protein, suggesting that posttranslational modification(s) may be necessary for optimal inhibitory activity. The tsetse thrombin inhibitor gene, which is present as a single copy in the tsetse genome, is expressed at high levels in salivary glands and midguts of adult tsetse flies, suggesting a possible role for the anticoagulant in both feeding and processing of the bloodmeal.

Jasmonate is essential for insect defense in Arabidopsis

The signaling pathways that allow plants to mount defenses against chewing insects are known to be complex. To investigate the role of jasmonate in wound signaling in Arabidopsis and to test whether parallel or redundant pathways exist for insect defense, we have studied a mutant (fad3–2 fad7–2 fad8) that is deficient in the jasmonate precursor linolenic acid. Mutant plants contained negligible levels of jasmonate and showed extremely high mortality (≈80%) from attack by larvae of a common saprophagous fungal gnat, Bradysia impatiens (Diptera: Sciaridae), even though neighboring wild-type plants were largely unaffected. Application of exogenous methyl jasmonate substantially protected the mutant plants and reduced mortality to ≈12%. These experiments precisely define the role of jasmonate as being essential for the induction of biologically effective defense in this plant–insect interaction. The transcripts of three wound-responsive genes were shown not to be induced by wounding of mutant plants but the same transcripts could be induced by application of methyl jasmonate. By contrast, measurements of transcript levels for a gene encoding glutathione S-transferase demonstrated that wound induction of this gene is independent of jasmonate synthesis. These results indicate that the mutant will be a good genetic model for testing the practical effectiveness of candidate defense genes.

Discovery of genes involved with learning and memory: An experimental synthesis of Hirschian and Benzerian perspectives

The biological bases of learning and memory are being revealed today with a wide array of molecular approaches, most of which entail the analysis of dysfunction produced by gene disruptions. This perspective derives both from early “genetic dissections” of learning in mutant Drosophila by Seymour Benzer and colleagues and from earlier behavior-genetic analyses of learning and in Diptera by Jerry Hirsch and coworkers. Three quantitative-genetic insights derived from these latter studies serve as guiding principles for the former. First, interacting polygenes underlie complex traits. Consequently, learning/memory defects associated with single-gene mutants can be quantified accurately only in equilibrated, heterogeneous genetic backgrounds. Second, complex behavioral responses will be composed of genetically distinct functional components. Thus, genetic dissection of complex traits into specific biobehavioral properties is likely. Finally, disruptions of genes involved with learning/memory are likely to have pleiotropic effects. As a result, task-relevant sensorimotor responses required for normal learning must be assessed carefully to interpret performance in learning/memory experiments. In addition, more specific conclusions will be obtained from reverse-genetic experiments, in which gene disruptions are restricted in time and/or space.

A novel application of gene arrays: Escherichia coli array provides insight into the biology of the obligate endosymbiont of tsetse flies

Symbiotic associations with microorganisms are pivotal in many insects. Yet, the functional roles of obligate symbionts have been difficult to study because it has not been possible to cultivate these organisms in vitro. The medically important tsetse fly (Diptera: Glossinidae) relies on its obligate endosymbiont, Wigglesworthia glossinidia, a member of the Enterobacteriaceae, closely related to Escherichia coli, for fertility and possibly nutrition. We show here that the intracellular Wigglesworthia has a reduced genome size smaller than 770 kb. In an attempt to understand the composition of its genome, we used the gene arrays developed for E. coli. We were able to identify 650 orthologous genes in Wigglesworthia corresponding to ≈85% of its genome. The arrays were also applied for expression analysis using Wigglesworthia cDNA and 61 gene products were detected, presumably coding for some of its most abundant products. Overall, genes involved in cell processes, DNA replication, transcription, and translation were found largely retained in the small genome of Wigglesworthia. In addition, genes coding for transport proteins, chaperones, biosynthesis of cofactors, and some amino acids were found to comprise a significant portion, suggesting an important role for these proteins in its symbiotic life. Based on its expression profile, we predict that Wigglesworthia may be a facultative anaerobic organism that utilizes ammonia as its major source of nitrogen. We present an application of E. coli gene arrays to obtain broad genome information for a closely related organism in the absence of complete genome sequence data.

Isolation and identification of a diuretic hormone from the mealworm Tenebrio molitor.

A diuretic hormone of unusual structure was isolated from extracts of whole heads of the mealworm Tenebrio molitor. The hormone is a 37-aa peptide of 4371 Da, with the sequence SPTISITAPIDVLRKTWEQERARKQMVKNREFLNSLN. This peptide increases cAMP production in Malpighian tubules of T. molitor. The amino acid sequence reveals that this peptide is a member of the family of sauvagine/corticotropin-releasing factor/urotensin I-related insect diuretic hormones. The C-terminal sequence of this peptide is quite different from other members of this family, which have a hydrophobic C terminus (isoleucinamide or valinamide). When aligned comparably, T. molitor diuretic hormone has a more hydrophilic C terminus, leucylasparagine (free acid). In contrast to all other known diuretic hormones of this family, this peptide has exceptionally low stimulatory activity on cAMP production in Malpighian tubules of Manduca sexta. However, at nanomolar concentrations it stimulates cAMP production in Malpighian tubules of T. molitor. Diuretic hormones of this family have been isolated previously from Lepidoptera, Orthoptera, Dictyoptera, and Diptera. This appears to be the first diuretic hormone isolated from a coleopteran insect.

Interazioni piante - insetti: ruolo delle formiche in un sistema multitrofico. Aspetti ecoetologici ed applicativi

Le formiche svolgono un importante ruolo all’interno degli ecosistemi ed alcune specie sono considerate keystone in quanto in grado di modificare la componente biotica e/o abiotica dell’ecosistema stesso. Sono animali ubiquitari che hanno colonizzato molteplici ambienti, compresi gli agroecosistemi. Negli agroecosistemi spesso svolgono un ruolo impattante determinando la diffusione o il regresso di specie di artropodi, alcune delle quali dannose alle colture. La presente ricerca tiene conto di un’ampia visione dei rapporti ecoetologici intercorrenti tra le formiche e la componente biotica di un ecosistema, utilizzando il concetto di rete multitrofica. In quest’ottica, si è pensato di costruire un sistema multitrofico costituito da una specie vegetale di interesse agrario (Cucumis sativus), dai suoi fitofagi naturali, divisi in fitomizi (afidi) (Aphis gossypii e Myzus persicae) e fitofagi masticatori (bruchi del lepidottero Mamestra brassicae), formiche (Formica pratensis) e predatori afidofagi (Aphidolets aphidimyza). Il sistema multitrofico è stato utilizzato sia per studiare l’aggressività delle formiche, sia per verificare l’esistenza di una comunicazione interspecifica tra le formiche e le piante (allelochimici). Gli studi sull’aggressività sono consistiti nel: • Verificare il livello di aggressività delle formiche nei confronti di un fitofago masticatore, competitore degli afidi nello sfruttare la pianta ospite. • Verificare se la presenza di afidi mutualisti fa variare il livello di aggressività delle formiche verso il competitore. • Verificare se esiste aggressività verso un predatore di afidi, i quali, secondo il paradigma della trofobiosi, dovrebbero essere difesi dalle formiche in cambio della melata. • Verificare se il predatore ha evoluto strategie volte ad eludere il controllo delle formiche sugli insetti che si approcciano alla colonia di afidi. Gli studi sui rapporti piante-formiche sono stati effettuati mediante olfattometro, osservando la risposta delle formiche alle sostanze volatili provenienti da piante infestate in modo differente con i fitofagi del sistema. Attraverso il trappolaggio e l’analisi gas-cromatografica delle sostanze prodotte dalle piante oggetto di studio abbiamo quindi individuato tipo e quantità di ogni composto volatile. Oltre alle piante di cetriolo, per questi esperimenti sono state utilizzate anche piante di patata (Solanum tuberosum). Dagli esperimenti sull’aggressività è risultato che le formiche manifestano un elevato potenziale predatorio, eradicando completamente la presenza dei bruchi sulle piante. Questo livello di aggressività tuttavia non cresce con la presenza degli afidi mutualisti che dovrebbero essere difesi dai competitori. Le formiche inoltre non sono in grado di sopprimere i predatori afidofagi che ipotizziamo riescano ad effettuare un camuffamento chimico, assumendo gli odori degli afidi dei quali si nutrono. I risultati degli esperimenti in olfattometro mostrano una chiara risposta positiva delle formiche verso gli odori di alcune delle piante infestate. Vi sono delle differenze nella risposta in funzione della specie di fitofago presente e della specie di pianta utilizzata. Nei trattamenti in cui erano presenti le piante di C. sativus, gli esperimenti in olfattometro hanno mostrato che le formiche rispondono in modo significativo agli odori emessi dalle piante in cui vi era la presenza del fitofago masticatore M. brassicae, solo o in associazione con A. gossypii. La presenza dei soli afidi, sia mutualisti (A. gossypii) sia non mutualisti (M. persicae), non ha invece indotto una risposta significativa nelle formiche rispetto agli odori delle piante non infestate. Nei trattamenti in cui erano presenti le piante di S. tuberosum la scelta delle formiche è stata significativa verso gli odori emessi dalle piante infestate con ciascuna delle singole specie di erbivori rispetto alle piante non infestate. Gli esperimenti sull’analisi delle sostanze volatili emesse dalle piante hanno confermato che gli organismi vegetali sono una vera centrale di produzione biochimica, infatti ben 91 composti volatili diversi sono stati individuati dall’analisi gas-cromatografica delle piante di cetriolo e 85 in quelle di patata. Dalle elaborazioni effettuate, rispettivamente 27 e 4 di essi sono prodotti esclusivamente dalle piante attaccate dai fitofagi. In generale, il cambiamento più consistente è dato dalla quantità di alcune sostanze volatili emesse dalle piante infestate rispetto a quelle integre che determina un cambiamento nei rapporti tra le sostanze che compongono i volatiles. E’ probabile che l’effetto attrattivo esercitato sulle formiche sia dato da un Blend di sostanze più che dai singoli composti presenti

Leishmaniose visceral americana: avaliação dos parâmetros da capacidade vetorial de Lutzomyia longipalpis em área urbana do muncípio de Panorama, São Paulo, Brasil

Introdução: A leishmaniose visceral (LV) é um importante problema de saúde pública no Brasil, com cerca 3000 mil casos notificados anualmente. Nos últimos anos, a LV tem ampliado sua distribuição em vários estados do país, associada principalmente aos processos socioambientais, antrópicos e migratórios. A LV é causada pela infecção com Leishmania infantum chagasi, transmitida, principalmente, por Lutzomyia longipalpis (Diptera: Psychodidae). Este flebotomíneo apresenta ampla distribuição nas Américas, todavia, evidências sugerem que se constitui em um complexo de espécies crípticas. A dinâmica de transmissão da LV é modulada por fatores ecológicos locais que influenciam a interação entre populações do patógeno, do vetor e dos hospedeiros vertebrados. Portanto, o estudo das variáveis associadas a esta interação pode contribuir para elucidar aspectos dos elos epidemiológicos e contribuir para a tomada de decisões em saúde pública. Objetivo: Avaliar parâmetros relacionados à capacidade vetorial da população de Lu. longipalpis presente em área urbana do município de Panorama, estado de São Paulo. Métodos: Foram realizadas capturas mensais durante 48 meses para avaliar a distribuição espaço-temporal de Lu. longipalpis e investigar a circulação de Le. i. chagasi. Também foram realizados os seguintes experimentos com o vetor: captura-marcação-soltura-recaptura para estimar a sobrevida da população e a duração do seu ciclo gonotrófico, a atratividade dos hospedeiros mais frequentes em áreas urbanas, a proporção de repasto em cão, infecção experimental e competência vetorial. Resultados: Observou-se que no município de Panorama, Lu. longipalpis apresentou as frequências mais elevadas na estação chuvosa (entre outubro e março), maior densidade em áreas com presença de vegetação e criação de animais domésticos, locais aonde também foi demonstrada a circulação natural de espécimes de Lu. longipalpis infectados com Le. i. chagasi. Além disto, foi corroborado que a população de Lu. longipalpis apresentou hábito hematofágico eclético, altas taxas de sobrevivência e que foi competente para transmitir o agente da LV. Nos experimentos de laboratório foi evidenciada a heterogeneidade na infecção de fêmeas de Lu. longipalpis desafiadas a se alimentarem em cães comprovadamente infectados por L. i. chagasi e o rápido desenvolvimento do parasita neste vetor natural. Conclusões. As observações do presente estudo corroboram a capacidade vetora de Lu. longipalpis para transmitir a Le. i. chagasi e ressaltam a importância da espécie na transmissão do agente etiológico da LV. Ações de manejo ambiental, educação e promoção à saúde são recomendadas às autoridades municipais para diminuir o risco potencial de infecção na população humana e canina, considerando-se o elevado potencial vetor de Lu. longipalpis e a presença de condições que favorecem a interação dos componentes da tríade epidemiológica da LV.

Influence of tree hollow characteristics on the diversity of saproxylic insect guilds in Iberian Mediterranean woodlands

Saproxylic diversity assessment is a major goal for conservation strategies in woodlands and it should consider woodland composition and configuration at site and tree level as key modelling factors. However, in Mediterranean woodlands little is known about the relation with the environmental factors that structure their assemblages, especially those linked to tree hollow microhabitats. We assessed the diversity of Syrphidae (Diptera) and Coleoptera saproxylic guilds that co-occurred in tree hollows located in three different Iberian Mediterranean woodlands in the Cabañeros National Park (Spain). Furthermore, we evaluated how differences in tree hollow microenvironmental variables (understood as the physical and biotic characteristics of a hollow and tree individual) influenced saproxylic guild diversity both within and among woodland sites. We found that woodland sites that provided greater heterogeneity of trees and hollow microhabitats determined higher saproxylic guild diversity. Nevertheless, certain species or even complete guilds can be favoured in woodlands where some hollow microhabitats predominate as a consequence of historical tree management. In general, hollow volume was the main determining factor for saproxylic guild richness and abundance in woodland sites, and large hollow volume was usually related to higher diversity, which highlighted the importance of multi-habitat hollow trees. Moreover, saproxylic guilds also responded to other different microenvironmental variables, which indicated different ecological preferences among guilds. The conservation of saproxylic insects in Iberian Mediterranean areas must be addressed to protect woodland sites that provide high diversity and large numbers of tree hollow microhabitats, and practices to enhance microhabitat heterogeneity should even be encouraged.

Where Is My Food? Brazilian Flower Fly Steals Prey from Carnivorous Sundews in a Newly Discovered Plant-Animal Interaction

A new interaction between insects and carnivorous plants is reported from Brazil. Larvae of the predatory flower fly Toxomerus basalis (Diptera: Syrphidae: Syrphinae) have been found scavenging on the sticky leaves of several carnivorous sundew species (Drosera, Droseraceae) in Minas Gerais and São Paulo states, SE Brazil. This syrphid apparently spends its whole larval stage feeding on prey trapped by Drosera leaves. The nature of this plant-animal relationship is discussed, as well as the Drosera species involved, and locations where T. basalis was observed. 180 years after the discovery of this flower fly species, its biology now has been revealed. This is (1) the first record of kleptoparasitism in the Syrphidae, (2) a new larval feeding mode for this family, and (3) the first report of a dipteran that shows a kleptoparasitic relationship with a carnivorous plant with adhesive flypaper traps. The first descriptions of the third instar larva and puparium of T. basalis based on Scanning Electron Microscope analysis are provided.

Insecta Saundersiana : or, Charcters of undescribed insects in the collection of William Wilson Saunders.

v.4 (1858) [Homoptera]

Katalog der Paläarktischen dipteren / bearbeitet von Th. Becker ... [et al.].

Bd. 2

Catalogus dipterorum hucusque descriptorum /

v. 3 (1908)

Katalog der Paläarktischen dipteren / bearbeitet von Th. Becker ... [et al.].

Bd. 4

Katalog der Paläarktischen dipteren / bearbeitet von Th. Becker ... [et al.].

Bd. 3

Catalogus dipterorum hucusque descriptorum /

v. 6 (1909)