Endocrine signatures underlying plasticity in postembryonic development of a lower termite, Cryptotermes secundus (Kalotermitidae)

Wood-dwelling termites are characterized by an extremely high and unique developmental flexibility that allows workers, which are immatures, to explore all caste options. The endocrine signatures underlying this flexibility are only vaguely understood. We determined juvenile hormone (JH) and ecdysteroid hemolymph titers during postembryonic development and in terminal instars of the drywood termite Cryptotermes secundus using field and laboratory colonies. Postembryonic development is characterized by a drop in JH titers at the transition from larval (individuals without wing buds) to nymphal (individuals with wing buds) instars. JH titers were low in winged sexuals and reproducing primary reproductives (< 200 pg/mu l) but were by an order of magnitude higher in neotenic replacement reproductives. The unique regressive molts of termites seem to be characterized by elevated JH titers, compared with progressive or stationary molts. Ecdysteroid titers were generally low in nymphal instars and in primary reproductives (< 50 pg/mu l). It was only during the third and fourth nymphal instars and in winged sexuals where some individuals showed elevated ecdysteroid titers. These results are the most comprehensive endocrinological data set available for any lower termite, with the potential to serve as baseline for understanding the extreme developmental flexibility underlying the evolution of social life in termites.

Molting dynamics and juvenile hormone titer profiles in the nymphal stages of a lower termite, Cryptotermes secundus (Kalotermitidae) - signatures of developmental plasticity

Termites are social cockroaches and this sociality is founded on a high plasticity during development. Three molting types (progressive, stationary and regressive molts) are fundamental to achieve plasticity during alate/sexual development, and they make termites a major challenge to any model on endocrine regulation in insect development. As the endocrine signatures underpinning this plasticity are barely understood, we studied the developmental dynamics and their underlying juvenile hormone OH) titers in a wood-dwelling termite. Cryptotermes secundus, which is characterized by an ancestral life style of living in dead wood and individuals being totipotent in development. The following general pattern elements could be identified during winged sexual development (i) regressive molts were accompanied by longer intermolt periods than other molting types, (ii) JH titers decreased gradually during the developmental transition from larva (immatures without wing buds), to nymph (immatures with wing buds), to winged adult, (iii) in all nymphal stages, the JH titer rose before the next molt and dropped thereafter within the first week, (iv) considerable variation in JH titers occurred in the midphase of the molting cycle of the 2nd and 3rd nymphal instar, inferring that this variation may reflect the underlying endocrine signature of each of the three molting types, (v) the 4th nymphal instar, the shortest of all, seems to be a switch point in development, as nymphs in this stage mainly developed progressively. When comparing these patterns with endocrine signatures seen in cockroaches, the developmental program of Cryprotermes can be interpreted as a co-option and repetitive use of hormonal dynamics of the post dorsal-closure phase of cockroach embryonic development. (C) 2012 Elsevier Ltd. All tights reserved.

Polyethism and comparability of termite choice assays in a model system using Microcerotermes turneri (Termitidae : Termitinae): Implications for standardised testing techniques

Traditional measures of termite food preference assess consequences of foraging behavior such as wood consumption, aggregation and/or termite survivorship. Although studies have been done to investigate the specifics of foraging behavior this is not generally integrated into choice assay experiments. Here choice assays were conducted with small isolated (orphaned) groups of workers and compared with choice assays involving foragers from whole nests (non-orphaned) in the laboratory. Aggregation to two different wood types was used as a measure of preference. Specific worker caste and instars participating in initial exploration were compared between assay methods, with samples of termites taken from nest carton material and sites where termites were feeding. Aggregation results differ between choice assay techniques. Castes and instars responsible for initial exploration, as determined in whole nest trials, were not commonly found exploring in isolated group trials, nor were they numerous in termites taken from active feeding sites. Consequently the use of small groups of M. turneri worker termites extracted from active feeding sites may not be appropriate for use in choice assays.

Genetic analysis of the breeding system of an invasive subterranean termite, Reticulitermes santonensis, in urban and natural habitats.

Reticulitermes santonensis is a subterranean termite that invades urban areas in France and elsewhere where it causes damage to human-built structures. We investigated the breeding system, colony and population genetic structure, and mode of dispersal of two French populations of R. santonensis. Termite workers were sampled from 43 and 31 collection points, respectively, from a natural population in west-central France (in and around the island of Oleron) and an urban population (Paris). Ten to 20 workers per collection point were genotyped at nine variable microsatellite loci to determine colony identity and to infer colony breeding structure. There was a total of 26 colonies, some of which were spatially expansive, extending up to 320 linear metres. Altogether, the analysis of genotype distribution, F-statistics and relatedness coefficients suggested that all colonies were extended families headed by numerous neotenics (nonwinged precocious reproductives) probably descended from pairs of primary (winged) reproductives. Isolation by distance among collection points within two large colonies from both populations suggested spatially separated reproductive centres with restricted movement of workers and neotenics. There was a moderate level of genetic differentiation (F(ST) = 0.10) between the Oleron and Paris populations, and the number of alleles was significantly higher in Oleron than in Paris, as expected if the Paris population went through bottlenecks when it was introduced from western France. We hypothesize that the diverse and flexible breeding systems found in subterranean termites pre-adapt them to invade new or marginal habitats. Considering that R. santonensis may be an introduced population of the North American species R. flavipes, a breeding system consisting primarily of extended family colonies containing many neotenic reproductives may facilitate human-mediated spread and establishment of R. santonensis in urban areas with harsh climates.

Inorganic elements in the fat bodies of Diatraea saccharalis (Lepidoptera: Crambidae) larvae parasitized by Cotesia flavipes (Hymenoptera: Braconidae)

Koinobiont parasitoids use several strategies to regulate the host`s physiological processes during parasitism. Although many aspects of host-parasitoid interactions have been explored, studies that attempted to assess the effects of parasitism on the availability of inorganic elements in the host are virtually nonexistent. Therefore, we aimed to evaluate the effects of parasitism on the concentrations of inorganic elements in the fat bodies of larvae of Diatraea saccharalis (Lepidoptera: Crambidae) during the development of the parasitoid Cotesia flavipes (Hymenoptera: Braconidae), by using total reflection X-ray fluorescence (TXRF). TXRF analysis allowed comparisons of the changes in the availability of the elements P. S. K, Ca, Cr, Fe, Ni, Cu, and Zn in the fat body tissues of D. saccharalis larvae parasitized by C. flavipes. Overall, the concentration of inorganic elements was higher early in parasitoid development (1 and 3 days after parasitism) compared to non-parasitized larvae, but much lower towards the end of parasitoid development (7 and 9 days after parasitism). Ca, K, and S were reduced after the fifth day of parasitism, which affected the total abundance of inorganic elements observed in the fat bodies of the parasitized hosts. The regulatory mechanisms or pathological effects related to the observed variation of the host inorganic elements induced by the parasitoid remain unknown, but there might be a strategy to make these elements available to the parasitoid larvae at the end of their development, when higher metabolic activity of the host fat body is required to sustain parasitoid growth. The observed variation of the host`s inorganic elements could also be related to the known effects of parasitism on the host`s immune response. (C) 2010 Elsevier Inc. All rights reserved.

Changes in the hemolymph and fat body metabolites of Diatraea saccharalis (Fabricius) (Lepidoptera : Crambidae) parasitized by Cotesia flavipes (Cameron) (Hymenoptera : Braconidae)

The koinobiont Cotesia flavipes responds to and is influenced by biochemical changes in the host hemolymph composition, Diatraea saccharalis. Changes in the composition of macronutrients may occur due to the hosts own development or by changes induced after parasitization. These changes occur to facilitate parasitoid invasion and to make the host internal environment suitable to parasitoid immature development. Therefore, changes in the availability of stored and circulating nutrients may correlate with the nutritional requirements of specific parasitoid immature stages. In here, we describe changes in the biochemical composition of parasitized and control larvae at different stages of parasitoid development to gain information on C flavipes host regulation and on its quantitative immature nutritional requirements. Total proteins, lipids and carbohydrates free in the hemolymph or stored in host fat bodies, and the SDS-PAGE protein profile of the hemolymph were evaluated in control and parasitized 6th instar during the whole parasitoid development. Changes in the total protein available in the host hemolymph were detected soon after parasitization, but carbohydrate and lipids were observed to differ only towards parasitoid larvae egression. Although C. flavipes affected the availability of all macronutrients observed in the host hemolymph, lipids and proteins stored in the host fat bodies were unaffected. However, carbohydrate concentration at the end of parasitoid larval development was much lower in parasitized than in control larvae at the same stage of development. SDS-PAGE analysis indicated C flavipes up-regulated two host proteins (125 and 48 kDa) and released two parasitism-specific proteins towards the end of parasitoid larval development. We provide a discussion on the role these changes may have on the process of host regulation and their possible requirement to sustain parasitoid development. (C) 2007 Elsevier Inc. All rights reserved.

Preferência de Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) por lagartas de Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) alimentadas com diferentes cultivares de cana-de-açúcar

O objetivo desta pesquisa foi avaliar a preferência do parasitoide Cotesia flavipes (Hymenoptera: Braconidae) por lagartas de Diatraea saccharalis (Lepidoptera: Crambidae), alimentadas com diferentes cultivares de cana-de-açúcar. O experimento foi conduzido em laboratório, em duas condições de alimentação, uma envolvendo lagartas de D. saccharalis, alimentadas em dieta artificial, e, a outra, com lagartas alimentadas em dieta artificial e mantidas temporariamente nos toletes dos cultivares de cana-de-açúcar. Os cultivares utilizados foram: SP80-1842 e SP81-3250, resistentes, e RB855536, suscetível à D. saccharalis. A preferência para oviposição das fêmeas de C. flavipes foi avaliada em testes com e sem chance de escolha, em delineamento inteiramente casualizado, com quatro tratamentos e 15 repetições. Lagartas de D. saccharalis com 19 dias de idade foram oferecidas para fêmeas de C. flavipes, acasaladas e com 24 horas de idade, em ambos os testes. No teste com chance de escolha, utilizou-se um olfatômetro, o qual constou de quatro compartimentos, em cujo centro liberaram-se quatro fêmeas de C. flavipes, enquanto, no teste sem chance de escolha, foram utilizadas placas de Petri, no interior das quais se colocou uma lagarta de D. saccharalis, oriunda das duas condições de alimentação. Nestes testes, foi observada a percentagem de parasitismo e, continua-mente, o comportamento da primeira escolha de fêmeas de C. flavipes, em intervalos de zero a um, um a três, três a cinco, cinco a oito e de oito a dez minutos após as liberações. Lagartas de D. saccharalis, oriundas da alimentação em dieta artificial com colmos triturados dos cultivares, foram igualmente preferidas para atratividade de fêmeas do parasitoide C. flavipes. A percentagem de parasitismo de lagartas de D. saccharalis, criadas com dietas artificiais com colmos dos cultivares SP80-1042 e RB855536, foi igualmente parasitadas por C. flavipes.

Novo gênero de Anacolini e descrição dos machos de Myzomorphus Flavipes e Poekilosoma Carinatipenne (Coleoptera, Cerambycidae, Prioninae)

The new genus Hovorelus and its type-species, H. splendidus sp. nov., are described from Peru. The males of Myzomorphus flavipes Galileo, 1987 and Poekilosoma carinatipenne Lane, 1941, both with sexual dimorphism, are described for the first time.

Comparison of the gut microbiota from soldier and worker castes of the termite Reticulitermes grassei

The bacterial microbiota from the whole gut of soldier and worker castes of the termite Reticulitermes grassei was isolated and studied. In addition, the 16S rDNA bacterial genes from gut DNA were PCR-amplified using Bacteria-selective primers, and the 16S rDNA amplicons subsequently cloned into Escherichia coli. Sequences of the cloned inserts were then used to determine closest relatives by comparison with published sequences and with sequences from our previous work. The clones were found to be affiliated with the phyla Spirochaetes, Proteobacteria, Firmicutes, Bacteroidetes, Actinobacteria, Synergistetes, Verrucomicrobia, and candidate phyla Termite Group 1 (TG1) and Termite Group 2 (TG2). No significant differences were observed with respect to the relative bacterial abundances between soldier and worker phylotypes. The phylotypes obtained in this study were compared with reported sequences from other termites, especially those of phylotypes related to Spirochaetes, Wolbachia (an Alphaproteobacteria), Actinobacteria, and TG1. Many of the clone phylotypes detected in soldiers grouped with those of workers. Moreover, clones CRgS91 (soldiers) and CRgW68 (workers), both affiliated with"Endomicrobia", were the same phylotype. Soldiers and workers also seemed to have similar relative protist abundances. Heterotrophic, poly-β-hydroxyalkanoate-accumulating bacteria were isolated from the gut of soldiers and shown to be affiliated with Actinobacteria and Gammaproteobacteria. We noted that Wolbachia was detected in soldiers but not in workers. Overall, the maintenance by soldiers and workers of comparable axial and radial redox gradients in the gut is consistent with the similarities in the prokaryotes and protists comprising their microbiota.

Spontaneous poisoning by larvae of Perreyia flavipes (Pergidae) in sheep

From a flock of 175 Texel sheep 25 animals died after consumption of a sawfly larvae subsequently identified as Perreyia flavipes. The disease occurred in June-July 2006 on a farm located in the county of Encruzilhada do Sul, Rio Grande do Sul, Brazil. Although there were 11 cattle in the same paddock, none of them was affected. High numbers of compact masses containing up to 150 larvae were scattered in the paddock where the animals were grazing. Most affected sheep showed severe apathy during 24-36 h before death, but weakness, muscular tremors and depression were also observed. Necropsy was performed on six sheep and the main macroscopic lesions were hemorrhages in the subcutaneous tissues, endocardium, gallbladder wall, and abomasal mucosa. In all animals was found hydrothorax, hydropericardium, ascites, and mild jaundice. Edema in the abomasal folds, mesentery, perirenal tissues, and gallbladder wall were also seen. The livers were yellowish with disseminated pinpoint hemorrhages in the parenchyma and had an enhanced lobular pattern. Perreyia flavipes larval body fragments and heads were found in the forestomach contents of the six sheep. Feces were scant, dry and formed balls coated by mucus and streaks of blood. Similar contents were also present at the end of the cecum. Prominent microscopic lesions included severe and diffuse periacinar or massive necrosis of hepatocytes associated with multifocal random hemorrhages. Diffuse necrosis of lymphoid follicles in lymph nodes and Peyer's patches, lymphoid depletion and necrosis in germinative centers of the spleen, and diffuse vacuolization in the renal tubular epithelia were also seen.

Intoxicação por larvas de Perreyia flavipes em bovinos na região sul do Rio Grande do Sul

Descrevem-se três surtos de intoxicação por larvas de Perreyia flavipes Konow, 1899 (Hymenoptera: Pergidae) ocorridos em bovinos durante os meses de julho e agosto de 2006. A morbidade foi de 0,8%, 6,2% e 33% nos três estabelecimentos, respectivamente. A letalidade foi de 100%. Os sinais clínicos nos três surtos caracterizaram-se por depressão, icterícia, decúbito com movimentos de pedalagem e morte em 24-48 horas. Macroscopicamente o fígado dos animais necropsiados estava aumentado de tamanho e com marcada acentuação do padrão lobular, os linfonodos hepáticos e mesentéricos estavam edemaciados e as placas de Peyer na mucosa do intestino delgado estavam deprimidas. Petequias e equimoses foram observadas no mesentério e abomaso. Histologicamente observou-se degeneração e necrose hemorrágica hepática que variou de centrolobular, se estendendo a região a mediozonal, ou massiva. Havia, ainda, marcada hemossiderose e necrose dos centros germinativos dos linfonodos, da polpa branca do baço e das placas de Peyer no intestino. A intoxicação ocorreu provavelmente em conseqüência da intensa seca observada na região nos meses de outubro a dezembro de 2005, período em que o inseto se encontra na forma de casulo enterrado no solo. A seca proporcionou maior emergência de adultos e conseqüentemente maior quantidade de posturas. A grande quantidade de matéria vegetal em decomposição devido às precipitações próximas do normal no verão proporcionou ambiente ideal para o desenvolvimento das larvas no período de inverno o que provavelmente levou à intoxicação.

Intoxicação espontânea por larvas de Perreyia flavipes (Pergidae) em suínos no estado de Santa Catarina

Descreve-se um surto de intoxicação espontânea por Perreyia flavipes em suínos. O surto ocorreu no final de maio de 2009, na cidade de Urubici, planalto serrano do Estado de Santa Catarina, Brasil. A propriedade tinha aproximadamente 50 animais criados extensivamente e desses 10 animais adoeceram. Esses suínos apresentavam anorexia, apatia, movimento constante de cabeça e bater de orelhas, dificuldade de caminhar, cambaleio, ranger de dentes e a agitação aumentava mediante ruídos e movimentos próximos. Na necropsia as alterações observadas foram a marcada evidenciação do padrão lobular hepático e a presença de larvas de P. flavipes misturadas ao conteúdo estomacal. Microscopicamente observou-se necrose de coagulação dos hepatócitos, com distribuição centrolobular a massiva que era acompanhada de congestão e hemorragia acentuada, restando uma ou duas camadas de hepatócitos com degeneração vacuolar na região portal. Os aspectos clínicos, epidemiológicos e as lesões caracterizaram hepatite tóxica por larvas de P. flavipes em suínos.

Perreyia flavipes larvae toxicity

Fresh or thawed Perreyia flavipes larvae were ground and mixed with water and orally ad ministered to sheep. At 5mg/kg, neither clinical nor enzymatic changes were observed. Unique do ses of 7.5 and 10mg/kg induced characteristic clinical signs of Perreyia sp. larvae poisoning, increased GGT and AST values, and decreased glycemic curves. However, doses of 5, 10, and 15mg/kg repeated at 30 or 15 days intervals caused no disease and mild disease followed by death, respectively. These fin dings indicate that these animals probably developed some degree of tolerance to the toxins in P. flavipes larvae. Ultrastru ctural examination of liver revealed proliferation of the smooth endoplasmic reticulum in the hepatocytes, which may be associated with an increased ability to metabolize toxins and could consequently lead to the tolerance observed in the present study. Further investigations may elucidate whether such tolerance effects could be applied as a control measure for P. flavipes poioning or other hepatotoxic diseases. In addition, clinicopathological findings were discussed.

Quality of Cotesia flavipes (Hymenoptera: Braconidae) reared at different host densities and the estimated cost of its commercial production

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fonte proteica na criação de Diatraea saccharalis e seu reflexo na produção e no controle de qualidade de Cotesia flavipes

O objetivo do trabalho foi avaliar diferentes dietas para lagartas de Diatraea saccharalis, com o intuito de obter informações sobre as características biológicas da praga e consequentemente avaliar a qualidade do parasitoide Cotesia flavipes, visando melhorar tecnicamente sua produção massal. Lagartas de 24 horas de idade foram transferidas para tubos (25 lagartas/tubo) contendo dieta artificial com levedura de cerveja e germe de trigo e outra somente com germe de trigo como fonte de proteína. Depois de aproximadamente 15 dias, as lagartas foram retiradas dos tubos e acondicionadas em placas com dieta, sendo observadas lagartas de D. saccharalis não parasitadas e parasitadas para avaliação das características biológicas de D. saccharalis e C. flavipes.nas diferentes dietas. Além da observação das características biológicas de D. saccharalis, foram realizadas medições nas lagartas de 15 dias de idade. Também, massas de pupas de C. flavipes.produzidas em biofábrica foram classificadas em três diferentes tamanhos, sendo pequenas (1,3 a 2,5 cm de comprimento), médias (2,5 a 3,5 cm de comprimento) e grandes (3,5 a 4,0 cm de comprimento). Após a emergência dos adultos foi realizado o parasitismo em lagartas de D. saccharalis, sendo observados o número de machos e fêmeas, tamanho da massa, razão sexual e número de pupas inviáveis. A dieta artificial que contém somente germe de trigo é a mais indicada para criação massal do hospedeiro D. saccharalis para produção em larga escala de C. flavipes. Massas de pupas classificadas como grandes possuem melhor qualidade em criações massais de C. flavipes.