A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombin coagulant inhibitory activity

Kazal-type inhibitors play several important roles in invertebrates, such as anticoagulant, vasodilator and antimicrobial activities. Putative Kazal-type inhibitors were described in several insect transcriptomes. In this paper we characterized for the first time a Kazal unique domain trypsin inhibitor from the Aedes aegypti mosquito. Previously, analyses of sialotranscriptome of A. aegypti showed the potential presence of a Kazal-type serine protease inhibitor, in female salivary glands, carcass and also in whole male, which we named AaTI (A. aegypti trypsin inhibitor). AaTI sequence showed amino acid sequence similarity with insect thrombin inhibitors, serine protease inhibitor from Litopenaeus vannamei hemocytes and tryptase inhibitor from leech Hirudo medicinalis (LDTI). In this work we expressed, purified and characterized the recombinant AaTI (rAaTI). Molecular weight of purified rAaTI was 7 kDa rAaTI presented dissociation constant (K(i)) of 0.15 and 3.8 nM toward trypsin and plasmin, respectively, and it weakly inhibited thrombin amidolytic activity. The rAaTI was also able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. AaTI transcription was confirmed in A. aegypti female salivary gland and gut 3 h and 24 h after blood feeding, suggesting that this molecule can act as anticoagulant during the feeding and digestive processes. Its transcription in larvae and pupae suggested that AaTI may also play other functions during the mosquito`s development. (C) 2010 Elsevier Masson SAS. All rights reserved.

The Brazilian 1994 stabilization plan: an analytical view

The Real Plan has succeeded in stabilizing the Brazilian inflation. The consumer price inflation has been reduced from 11260 percent per year, in June 1994, to an estimate of 8 percent in 1997. The lower inflation resulted in a remarkable income distribution, and in an increased private consumption. The plan managed to control the inflationary effects of the increased demand with some traditional measures: A more liberalized economy, a moving (and overvalued) exchange rate band, high interest rate differentials, and a tight domestic credit policy. The government has, so far failed to accomplish the fiscal adjustment. The price stabilization has largely depended on the current account deficit. However, macroeconomic indicators do not present reasons for concern about the current account sustainability, in the medium-run. The economy may be trapped in a low-growth vicious cycle, represented by a stop-and-go trend, due to the two-way endogencity between domestic saving and growth. Economic growth depends on policies in increase the public sector saving, to secure the privatization of the State enterprises, and to promote investments. The major problem for the government action is, as always, in the political sphere. Approximately 80 percent of the Central Government net revenue are allocated to the social sectors. Consequently, the fiscal reform will hue to deal with the problem of re-designing the public sector’s intervention in the social area. Most probably, it will be inevitable to cut the social area budget. This is politically unpleasant.

Abundância e distribuição sazonal de espécies autóctones de camarões peneídeos no canal de abastecimento do sistema primar de aquicultura orgânica (Laguna de Guaraíras, Tibau do Sul, Rio Grande do Norte, Brasil)

Guaraíras lagoon, located in Tibau do Sul in the eastern littoral of Rio Grande do Norte (Brazil), presents a permanent connection to the sea, which guarantees the occurrence of a rich biodiversity, which includes the autochthonous shrimp species Litopenaeus schmitti, Farfantepenaeus subtilis and Farfantepenaeus brasiliensis. In spite of being subject to a strong human intervention in the last decade, mainly related to the installation of shrimp (Litopenaeus vannamei) farms, the lagoon is still scarcely studied. The present study aims at characterizing the populations of the three autochthonous penaeid shrimp species inhabiting Guaraíras, taking into consideration their abundance and seasonal distribution in the inflow channel of Primar System of Organic Aquaculture (Tibau do Sul, Rio Grande do Norte, Brazil). Twelve monthly samples were carried out from May 2005 to April 2006 with the aid of a circular cast net in the inflow channel, which is daily supplied with water from Guaraíras. Sampling months were grouped in trimesters according to the total pluviosity, thus comprising four trimesters. Water salinity was monitored twice a week and temperature values registered on a daily basis at noon, during the study period. The daily pluviosity data from the municipality of Tibau do Sul were supplied by Empresa de Pesquisa Agropecuária do Rio Grande do Norte (EMPARN). Collected shrimp were identified, weighted, measured and sexed. L. schmitti specimens (0.2 g to 17.8 g) were distributed in 1.3 g weight classes intervals. From the eighth sampling month (December 2005) onwards, males were classified into three categories, in accordance with the development of their petasm: (a) rudimentary petasm, (b) partially formed petasm, and (c) completely formed petasm. Among the ecological variables, rainfall showed the greatest dispersion (s.d.=187.74Rainfall and abundance of L. schmitti were negatively correlated (r = -0.85) whereas its abundance and water salinity were positively correlated (r = 0.63). Among 1,144 collected individuals, 1,127 were L. schmitti, 13 were F. subtilis and 4 were F. brasiliensis, which corresponded to 98.51%, 1.14% and 0.35% of the total of collected individuals. L. schmitti occurred in 100 % of all samples. Differently, the presence of F. subtilis and F. brasiliensis was restricted to 33% and 17% of the collected samples, respectively. The present study confirmed the occurrence of L. schmitti, F. brasiliensis and F. subtilis in Guaraíras. However, this lagoon seems to be primarily inhabited by juvenile Litopenaeus schmitti. The population of L. schmitti analysed showed a seasonal pattern of distribution. In general, in the months of high salinity and absence of rain, the number of individuals was higher than in the wet months. Further studies on the reproductive biology and ecology of L. schmitti, F. brasiliensis and F. subtilis may elucidate questions referring to the abundance, period, and phase of occurrence of these shrimp genera in Guaraíras. Finally, the risks associated to the establishment of L. vannamei in the lagoon provide a novel outlet for studies in this biotope

Avaliação da toxicidade de florações naturais e de cultura de cianobactérias: efeitos sobre Ceriodaphnia silvestri (Crustacea, Cladocera)

The incidence of toxic cyanobacterial blooms is one of the important consequences of eutrophication in aquatic ecosystems. It is a very common phenomenon in reservoirs and shrimp ponds in the State of Rio Grande do Norte (RN), Brazil. Cyanobacterias produce toxins which can affect aquatic organisms and men trough the food chain. Aiming to contribute to the studies of cyanobacterias in RN, we propose: a) to evaluate the toxicity of isolated cyanobacterias in important fresh-water environments; and b) to verify the effects of both natural and cultured blooms occurred in reservoirs for human supply and in the cladoceran Ceriodaphnia silvestrii. This study was carried out using samples of natural blooms occurred between March and October of 2004 in Gargalheiras Dam (08º L e 39º W), in July of 2004 in Armando Ribeiro Gonçalves Dam (06o S e 37o W) and in commercial shrimp ponds (Litopenaeus vannamei) located in fresh-water environments. The samples were collected with plankton net (20µm.) for identification, isolation and obtaining of phytoplanktonic biomass for liophilization and later toxicity bioassays. The toxicity of cultured samples and natural blooms was investigated through bioassays in Swiss mice. Quantification of cyanobacteria in samples was conducted following the Ütermol method, with 300mL samples fixed with lugol. The toxicity test with Ceriodaphnia silvestrii followed ABNT, 2001 recommendations, and were accomplished with natural hepatotoxic bloom s samples and cultured samples of both non-toxic and neurotoxic C. raciborskii. In this test, five newborns, aged between 6 and 24 hours, were exposed to different concentrations (0 a 800 mg.L-1) of crude cyanobacterial extracts during 24 and 48 hours. Three replicates were used per treatment. The pH, temperature and dissolved oxygen at the beginning and after 24 and 48hours from the test were measured. We estimated the CL50 through the Trimmed Spearman-Karber method. The blooms were constituted by Microcystis panniformis, M. aeruginosa, Anabaena circinalis, Cylindrospermopsis raciborskii and Planktothrix agardhii, producers of mycrocistin-LR confirmed with HPLC analysis. Samples of hepatotoxic blooms registered toxinogenic potential for C. silvestrii, with CL50-24h value of 47.48 mg.L-1 and CL5048h of 38.15 mg.L-1 for GARG samples in march/2005; CL50-24h of 113,13 mg.L-1 and CL5048h of 88,24 mg.L-1 for ARG July/2004; CL50-24h of 300.39 mg.L-1 and CL50-48h of 149.89 mg.L-1 for GARG October/2005. For cultured samples, values of CL50-24h and CL50-48h for C. raciborskii toxic strains were 228.05 and 120.28 mg.L-1, respectively. There was no mortality of C. silvestrii during the tests with non-toxic C. raciborskii strain. The toxicity test with C. silvestrii presented good sensitivity degree to cyanotoxins. The toxicity of natural hepatotoxic blooms samples (microcystins) and cultured neurotoxic saxitoxins producer samples analyzed in this study give us strong indications of that toxin s influence on the zooplanktonic community structure in tropical aquatic environments. Eleven cyanobacteria strains were isolated, representing 6 species: Anabaenopsis sp., Cylindrospermopsis raciborskii, Chroococcus sp., Microcystis panniformis, Geitlerinema unigranulatum e Planktothrix agardhii. None presented toxicity in Swiss mice. The strains were catalogued and deposited in the Laboratório de Ecologia e Toxicologia de Organismos Aquáticos (LETMA), in UFRN, and will be utilized in ecotoxicológical and ecophysiological studies, aiming to clarify the causes and control of cyanobacterial blooms in aquatic environments in RN. This state s reservoirs must receive broader attention from the authorities, considering the constant blooms occurring in waters used for human consumption

Validação de metodologia para análise de aminas bioativas em camarão utilizando cromatografia de íons

In this study, we worked with the validation of a methodology for analysis of bioactive amines in shrimp, considering it to be one of the main products of the northriograndense trade balance, maintaining the state of Rio Grande do Norte topped the list of Brazilian exports of this product the last decade. The sector of the Brazilian shrimp works exclusively with gray shrimp Litopenaeus Vannamei since the late 1990s. This study used liquid chromatography with conductimetric detector, using as the mobile phase methylsulfonic 3 mM acid (MSA) with gradient and phase C18 column with reverse the development of methodology for the analysis of bioactive amines in shrimp. In the sample preparation was used as 5% trichloroacetic acid (TCA) extraction solution. Validation analysis of biotativas amines (putrescine - PUT, histamine - HIST, agmatine - AGM, spermidine - EPD and spermine - EPN) in shrimp, the linear working range was 0.1 to 2.0 mg L-1 to was sensitive, homoscedastic, in effect, selective, accurate and precise array. Thus, considered feasible for these determinations bioactive amines in this array. Determined the concentration of these amines in fresh shrimps (AGM = 0.61 ± 0.05 mg kg- 1 EPD = 2.57 ± 0.14 mg kg-1 and EPN = 1.79 ± 0.11 mg kg-1), and freezing weather predetermined in cooked shrimp (AGM = 6.28 ± 0.18 mg kg-1, EPD = 12.72 ± 0.02 mg kg- 1 and EPN = 22.30 ± 0.60 mg kg-1), the shrimp with twenty-four hour stay at room temperature (PUT = 879.52 ± 28.12 mg kg-1, AGM = 848.13 ± 19.40 mg kg-1, ESPD = 13.59 ± 0.97 mg kg-1 and ESPN = 18.47 + 1.57 mg kg-1). In shrimp subjected to freezing for a week, two weeks, three weeks and four weeks, the results showed that there is an increase in the content of agmatine (7.31 ± 0.21 mg kg-1) while in spermine ( 1.22 ± 0.14 mg kg-1) and spermidine (below limit of quantification) there was a decrease in the freeze time, while there is a decrease in the level of spermidine not reaching detectad. The putrescine was only found in shrimp that remained for 24 hours at room temperature and histamine was not found in any of the samples

Avaliação de variante somaclonal de porte baixo de bananeira 'Nanicão Jangada' (Musa sp) em duas densidades

Avaliou-se, sob duas densidades de plantio, um variante somaclonal de porte baixo de bananeira, comparando-o com a variedade Nanicão Jangada que lhe deu origem. Os materiais genéticos 'Nanicão Jangada'(controle) e o variante somaclonal representado pelas seleções 224 e 225 de um ensaio anterior, foram avaliados nos espaçamentos 2,0m X 2,0m (densidade 2500 plantas ha-1) e 3,0m X 2,0m (1666plantas ha-1), na Fazenda de Ensino e Pesquisa da Faculdade de Engenharia - UNESP, Campus de Ilha Solteira-SP. O ensaio foi conduzido em blocos ao acaso com cinco repetições, com utilização de mudas micropropagadas, sob irrigação por gotejamento, no período de dezembro de 1998 a março de 2001, com avaliação dos dois primeiros ciclos de produção. Constatou-se efeito da densidade e do ciclo sobre a produção estimada de frutos sendo que no cultivo mais denso, a média foi de 81,25 t.ha-1 no primeiro ciclo de produção e 67,93 t.ha-1 no segundo ciclo. No cultivo de menor densidade a produção estimada no primeiro ciclo foi de 51,35 t.ha-1 e 44,08 t.ha-1 no segundo. As seleções do variante de porte baixo apresentaram menor altura da planta e mostraram-se relativamente mais precoces e com produção semelhante a cv. Nanicão Jangada no primeiro ciclo. No segundo ciclo houve uma queda na produção do bananal, com maior intensidade para a seleção 225 do variante.

Stochastic dynamics in exotic and native blowflies: an analysis combining laboratory experiments and a two-patch metapopulation model

In this study we explored the stochastic population dynamics of three exotic blowfly species, Chrysomya albiceps, Chrysomya megacephala and Chrysomya putoria, and two native species, Cochliomyia macellaria and Lucilia eximia, by combining a density-dependent growth model with a two-patch metapopulation model. Stochastic fecundity, survival and migration were investigated by permitting random variations between predetermined demographic boundary values based on experimental data. Lucilia eximia and Chrysomya albiceps were the species most susceptible to the risk of local extinction. Cochliomyia macellaria, C. megacephala and C. putoria exhibited lower risks of extinction when compared to the other species. The simultaneous analysis of stochastic fecundity and survival revealed an increase in the extinction risk for all species. When stochastic fecundity, survival and migration were simulated together, the coupled populations were synchronized in the five species. These results are discussed, emphasizing biological invasion and interspecific interaction dynamics.

The invasion of five alien species in the Delta do Parnaiba Environmental Protection Area, Northeastern Brazil

Marine biological invasions have been regarded as one of the major causes of native biodiversity loss, with shipping and aquaculture being the leading contributors for the introductions of alien species in aquatic ecosystems. In the present study, five aquatic alien species (one mollusk, three crustaceans and one fish species) were detected during dives, shore searches and from the fisheries on the coast of the Delta do Parnaiba Environmental Protection Area, in the States of Piaui and Maranhao, Northeastern Brazil. The species were the bicolor purse-oyster Isognomon bicolor, the whiteleg shrimp Litopenaeus vannamei, the giant river prawn Macrobrachium rosenbergii, the Indo-Pacific swimming crab Charybdis hellerii and, the muzzled blenny Omobranchus punctatus. Ballast water (I. bicolor, C. hellerii, and O. punctatus) and aquaculture activities (L. vannamei and M. rosenbergii) in adjacent areas are the most likely vectors of introduction. All exotic species found have potential impact risks to the environment because they are able to compete against native species for resources (food and habitat). Isognomon bicolor share the same habitat and food items with the native bivalve species of mussels and barnacles. Litopenaeus vannamei share the same habitat and food items with the native penaeids such as the pinkspot shrimp Farfantepenaeus brasiliensis, the Southern brown shrimp Farfantepenaeus subtilis, and the Southern white shrimp Litopenaeus schmitti, and in the past few years L. vannamei was responsible for a viral epidemics in the cultivation tanks that could be transmitted to native penaeid shrimps. Charybdis hellerii is also able to cause impacts on the local fisheries as the species can decrease the populations of native portunid crabs which are commercialized in the studied region. Macrobrachium rosenbergii may be sharing natural resources with the Amazon River prawn Macrobrachium amazonicum. Omobranchus punctatus shares habit with the native redlip blenny Ophioblennius atlanticus and other fishes, such as the frillfin goby Bathigobius soporator. Some immediate remedial measures to prevent further introductions from ballast water and shrimp farm ponds should be: (i) to prevent the release of ballast water by ship/vessels in the region; (ii) to reroute all effluent waters from shrimp rearing facilities through an underground or above-ground dry well; (iii) to install adequate sand and gravel filter which will allow passage of water but not livestock; (iv) outdoor shrimp pounds located on floodable land should be diked, and; (v) to promote environmental awareness of those directly involved with ballast water (crews of ship/vessels) and shrimp farms in the region. Rev. Biol. Trop. 58 (3): 909-923. Epub 2010 September 01.

Ciclo reprodutivo do caranguejo violinista Uca rapax (Smith) (Crustacea, Brachyura, Ocypodidae) habitante de um estuário degradado em Paraty, Rio de Janeiro, Brasil

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

Epibiosis in decapod crustaceans by stalked barnacle Octolasmis lowei (Cirripedia: Poecilasmatidae)

Stalked barnacles Octolasmis lowei Darwin, 1851 are frequently found attached to decapod crustaceans. Their epibiotic association depends on many factors, which are mainly related to characteristics of the host's biology. This study evaluated the infestation and distribution of stalked barnacles in the branchial chambers of crabs, and analyzed the data with respect to the host's sex, maturity stage, molt cycle and size. The crab species Arenaeus cribrarius Lamarck, 1818, Callinectes danae Smith, 1869, Callinectes ornatus Ordway, 1863, Hepatus pudibundus Herbst, 1785, Libinia ferreirae Brito Capello, 1871, and Persephona punctata Linnaeus, 1758 were sampled and found to be infested by O. lowei. No juvenile crabs were infested. The prevalence of infestation by O. lowei was significantly different among C. danae, C. ornatus, and H. pudibundus males and females. All infested hosts were in the intermolt period. The mean size of infested crabs was larger than that observed for non-infested individuals. Internally, stalked barnacles were concentrated on the central gills or walls and floor of branchial chambers, suggesting that these gills provide more favorable conditions for the settlement and development of these epibionts. These results highlight the relationship between epibiont infestation and host biology, as well as the role of decapod crustaceans as a suitable substrate for the development of stalked barnacle O. lowei. © 2013 Sociedade Brasileira de Zoologia All rights reserved.

Crescimento inicial de um clone de Eucalyptus grandis em diferentes arranjos de plantio no sistema de curta rotação

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

Policultivo de acará-bandeira e camarão-marinho

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Ecofisiologia de espécies ocorrentes em uma floresta de restinga

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Regeneração florestal associada a tamanhos de clareiras: implicações para o manejo florestal sustentável

A crença na capacidade de regeneração florestal é um dos principais sustentáculos da concepção de manejo madeireiro sustentável em longo prazo. O desempenho do processo regenerativo, por sua vez, depende da intensidade dos danos causados pela atividade madeireira, os quais podem ser reduzidos desde que se disponha de dados sistemáticos que orientem critérios adequados ao bom manejo florestal. O presente estudo, realizado em Paragominas, Pará, tem como objetivo avaliar como o tamanho das clareiras afeta a regeneração florestal. Para efetivar essa avaliação, foram monitorados elos do processo regenerativo (e. g., herbívoros vertebrados, chuva de sementes, fatores físicos) e/ou vários atributos diretos da regeneração (e. g., densidade, riqueza, crescimento, recrutamento, mortalidade de plantas) em dois sítios do referido município. Na Fazenda Rio Capim, com exploração recente, quinze clareiras com idade de 1,3 ano foram selecionadas em cerca de 300 ha de floresta explorada sob impacto reduzido e monitoradas durante 15 meses. As clareiras compreenderam três categorias de tamanho: 05 pequenas (30-100 m2), 05 médias (500-800 m2) e 05 grandes (> 1.500 m2). Na Fazenda Cauaxi, com exploração antiga, somente os atributos diretos da regeneração foram avaliados em doze clareiras com 8,5 anos de idade, sendo quatro de cada categoria de tamanho acima mencionada, exceto as clareiras grandes que foram menores (1.000-1.400 m2). A hipótese geral deste estudo é que o comportamento dos diversos fatores analisados favorecerá maior riqueza de espécies em regime de distúrbios intermediários, neste caso, em clareiras médias (sensu Connell, 1978). De modo geral, essa hipótese não foi corroborada. Na Fazenda Rio Capim (1,3 ano pós-exploração), apesar das clareiras grandes terem sido significativamente mais pobres em espécies do que todos os demais ambientes, as clareiras médias não foram aquelas que apresentaram maior riqueza. Dentre os tamanhos de clareiras analisados, as clareiras grandes foram as que mais se diferenciaram da condição controle (floresta fechada), apresentando maiores temperaturas, maior densidade e crescimento da regeneração e maior taxa de crescimento de cipós. Nas clareiras médias, os cipós e espécies pioneiras também cresceram significativamente mais rápido do que nas clareiras pequenas e floresta fechada. As clareiras pequenas foram mais semelhantes à floresta fechada, diferindo apenas por sua maior densidade de indivíduos de espécies pioneiras e maior taxa de crescimento da regeneração (exceto cipós). A chuva de sementes e o impacto de mamíferos herbívoros sobre a regeneração foram indiferentes ao tamanho das clareiras, mas mostraram-se dependentes de características pontuais, como presença de fontes alimentares para atrair fauna e fornecer sementes. As clareiras antigas (8,5 anos) da Fazenda Cauaxi não apresentaram nenhuma divergência significativa entre si nem com a amostra controle. Comparativamente com as clareiras jovens, as clareiras antigas denotaram menor densidade e maior riqueza relativa. Considerando-se todas as diferenças observadas entre os diferentes tamanhos de clareiras e floresta fechada e suas potenciais implicações sobre o processo regenerativo, recomenda-se que grandes clareiras sejam evitadas. As clareiras pequenas e médias reúnem mais atributos favoráveis à sustentabilidade do manejo madeireiro.

Avaliação de dietas lipídicas fornecidas em duas frequências de suplementação na terminação de novilhas em pastejo

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)