Utilização de embriões liofilizados e flocos de Artemia na diversificação nutricional de pós-larvas do camarão marinho Litopenaes vannamei (Boone, 1931)

The objective of the current study was to evaluate the zootechnical performance (survival and growth) of Litopenaeus vannamei post-Iarvae fed an artificial shrimp diet supplemented with Artemia flakes or freeze-dried Artemia embryos. For that purpose, 20 culturing units were individually stocked with 50 shrimp post-Iarvae (average dry weight of 0,3 ± 0,03 mg) at a stocking density of 20 post-larvae per liter, and fed the experimental diets to satiation during 20 days. The experimental design consisted of four diets (T1, T2, T3 and T4) with five repetitions each. For treatments T1, T2 and T3, dietary supplements of 5mg of Artemia flakes (T1), freeze-dried Artemia embryos (T2), and of the commercial shrimp diet (T3) were offered 2 hours after the shrimp were initially fed the commercial shrimp diet. For treatment T4 (control), no additive was offered 2 hours after the initial feeding. Shrimp survival, absolut (GPA) and relative increase in weight (GPR), and specific growth rate (TCR) were used as evaluation criteria. After the experimental period, no significant statistical differences (p>0,05) in survival were observed. Regarding growth, the dietary treatment which used freeze-dried Artemia embryos as an additive (T2) presented the best results for GPA (6,7 ± 0,7 mg). There were no statistical differences within treatments T1, T3 and T4 (p>0,05). AIso, post-larvae fed freeze-dried embryos (T2) showed a relative increase in weight (2241,4%) which differed significantly (p<0,05) from T4(1911,7%) but not from T1 (1801,6%) or T3 (1946,7%). In conclusion, the results of the current study indicate that an artificial shrimp diet supplemented with freeze-dried Artemia embryos fulfils the nutritional requirements of post-larvae L. vannamei and promotes a better growth than diets not supplemented with Artemia flakes

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Potencial larvicida de extratos de plantas regionais no controle de larvas de Aedes aegypti

Dengue, amongst the virus illnesses one can get by vectorial transmission, is the one that causes more impact in the morbidity and mortality of world s population. The resistance to the insecticides has caused difficulties to control of vector insect (Aedes aegypti) and has stimulated a search for vegetables with larvicidal activity. The biodiversity of Caatinga is barely known and it is potential of use even less. Some plants of this biome are commercialized in free fairs northeast of Brazil, because of its phytotherapics properties. The vegetables in this study had been selected by means of a questionnaire applied between grass salesmen and natives of the Serido region from Rio Grande do Norte state; culicids eggs had been acquired with traps and placed in container with water for the larva birth. Thirty larvae had been used in each group (a group control and five experimental groups), with four repetitions four times. The vegetables had been submitted to the processes of decoction, infusion and maceration in the standard concentration of 100g of the vegetable of study in 1l of H2O and analyzed after ½, 1, 2, 4, 8, 12, 24 and 48 hours for verification of the average lethal dose (LD50) from the groups with thirty larva. The LD50 was analyzed in different concentrations (50g/l, 100g/l, 150g/l, 200g/l e 300g/l) of Aspidosperma pyrifolium Mart. 48 extracts of rind, leaf and stem of the seven vegetal species: Aspidosperma pyrifolium Mart., Mimosa verrucosa Benth, Mimosa hostilis (Mart.) Benth., Myracrodruon urundeuva Allemão, Ximenia americana L, Bumelia sartorum Mart Zizyphus joazeiro Mart, had been analyzed. The extracts proceeding from the three methods were submitted to the freezedrying, to evaluate and to quantify substances extracted in each process. The results had shown that Aspidosperma pyrifolium Mart. and Myracrodruon urundeuva Allemão are the species that are more distinguished as larvicidal after 24 hours of experiment, in all used processes of extraction in the assays. The Zizyphus joazeiro Mart species has not shown larvicidal activity in none of the assays. In relation to the extraction method, the decoction was the most efficient method in the mortality tax of the A. aegypti larvae

Parâmetros parasitológicos e desempenho produtivo de ovinos naturalmente infectados com nematoides gastrintestinais mantidos em diferentes pastagens tropicais

The control of gastrointestinal nematodes of small ruminants is usually done with anthelmintics. However, due to the emergence of ever-increasing parasite resistance to these drugs, looking up an alternative control parasites. One of this is sought in pasture management, as these are the sources of animals` infection by L3 infective larvae of helminths. The aim of this study was to evaluate the performance of sheep to natural infections by gastrointestinal nematodes grazing Panicum maximum cv. Massai and cv. Aruana, and Brachiaria brizantha cv. Piatã and cv. Marandu. The work was conducted from May to August-2011 with 48 male sheeps SRD versus Santa Inês breed. The animals were naturally infected with gastrointestinal nematodes, and maintained in four different cultivars of tropical forage grasses, naturally contaminated with eggs and larvae of gastrointestinal nematodes. Each week the animals were phenotypically characterized by parasitological examinations (eggs per gram of feces - EPG, and feces culture), hematological (packed cell volume PCV, and blood eosinophil count) method to evaluate the Famacha© colorof ocular mucosa, and the measures of body condition score and weight. In pastures was made the recovery of infective larvae in order to determine the quantity of L3 present in the pasture. The experimental design was a randomized completed block with two replications and before the entry of animals in the paddocks, they have been wormed. The experiment was ended when the animals reached 32.0 kg liveweight, and then were slaughtered and autopsies performed for the recovery and identify parasites of the abomasum, small intestine and large intestine. Results were subjected to analysis of variance, t test and Tukey`s test. The animals kept on pastures of Marandu grass had lower EPG counts, higher percentage of packed cell volume and higher average weight; those who remained in the Piatã pasture had lower eosinophil counts per microliter of blood. About the Famacha©, the highest prevalence was Famacha 2, and the body condition score ranged between 2 and 3. The results of feces cultures and recovery of larvae on pastures showed the presence of larvae of Trichostrongylus sp., and at the necropsy too. This way, it was concluded that the grass cultivars influences the sheep parasite load; the Famacha, together with EPG and packed cell volume are important indicators for use in controlling gastrointestinal nematode infections. The Trichostrongylus sp. was the most prevalent parasite in sheep during the rainy season

Resposta de ovinos naturalmente infectados por nematoides gastrintestinais em pastos de capim-massai

The aim of this study was to evaluate the crossbred sheep Santa Ines response to natural infections by gastrointestinal nematodes in Panicum maximum cv. pastures Massai at different times of grazing. For this it was used 36 crossbred sheep Santa Inês, divided into four groups and randomly distributed in massai-grass pickets naturally infected by gastrointestinal nematodes. In the period from October 2013 to May 2014, the animals were weekly monitored for hematology tests (packed cell volume) and parasitological (egg counts per gram of feces and fecal culture) in order to monitor the level of infection of sheep and they were assessed for weight, body condition score and staining ocular mucosa by FAMACHA© method. Furthermore infective larvae was recovery from pasture in times of pre-and post-grazing pasture. At 35 kg body weight, the animals were slaughtered and necropsied to collect the contents of the abomasum and small and large intestines to recover the adult parasites, carrying out the count and identification. The animals on pasture with a height of 40 cm pre-grazing obtained low nematode eggs in feces (1608 eggs / g) and 33 cm those in pre-grazing the highest average (2,539 eggs / g). The animals in different groups showed mean values of packed cell volume less than 23% (P> 0.05). Regarding to FAMACHA, the animals belonging to the groups 40 and 50 cm pre-grazing remained over 50% of individuals between FAMACHA 1 and 2 , and those to 33 and 45 cm pre-grazing showed mostly between FAMACHA 3, 4 and 5 (69.06% and 58.93%, respectively). In the culture of larvae in feces and recovery of pasture larvae before and after grazing the following genera were found, in order of prevalence: Haemonchus, Trichostrongylus, Oesophagostomum and Strongyloides. The pasture with 33 cm of pre-grazing provided the highest number of larvae recovered both in the pre and post-grazing (1,081 and 715 L3 / 100 g of green matter, respectively). Among the different groups, the sheep had an average weight of between 23 and 26 kg and variation between the body 1 and 3 scores with scores greater frequency of 1.5 to 2.It can be observed that the animals kept at different times of grazing got different answers on the parasite load. Animals kept in 40 cm pre-grazing expressed a satisfactory answer than the other groups (33, 45 and 50 cm), best supporting the action of endoparasites. Moreover, this same height, was recovered fewer infective larvae on pasture in pre and post-grazing.

Utilização de embriões liofilizados e flocos de Artemia na diversificação nutricional de pós-larvas do camarão marinho Litopenaes vannamei (Boone, 1931)

The objective of the current study was to evaluate the zootechnical performance (survival and growth) of Litopenaeus vannamei post-Iarvae fed an artificial shrimp diet supplemented with Artemia flakes or freeze-dried Artemia embryos. For that purpose, 20 culturing units were individually stocked with 50 shrimp post-Iarvae (average dry weight of 0,3 ± 0,03 mg) at a stocking density of 20 post-larvae per liter, and fed the experimental diets to satiation during 20 days. The experimental design consisted of four diets (T1, T2, T3 and T4) with five repetitions each. For treatments T1, T2 and T3, dietary supplements of 5mg of Artemia flakes (T1), freeze-dried Artemia embryos (T2), and of the commercial shrimp diet (T3) were offered 2 hours after the shrimp were initially fed the commercial shrimp diet. For treatment T4 (control), no additive was offered 2 hours after the initial feeding. Shrimp survival, absolut (GPA) and relative increase in weight (GPR), and specific growth rate (TCR) were used as evaluation criteria. After the experimental period, no significant statistical differences (p>0,05) in survival were observed. Regarding growth, the dietary treatment which used freeze-dried Artemia embryos as an additive (T2) presented the best results for GPA (6,7 ± 0,7 mg). There were no statistical differences within treatments T1, T3 and T4 (p>0,05). AIso, post-larvae fed freeze-dried embryos (T2) showed a relative increase in weight (2241,4%) which differed significantly (p<0,05) from T4(1911,7%) but not from T1 (1801,6%) or T3 (1946,7%). In conclusion, the results of the current study indicate that an artificial shrimp diet supplemented with freeze-dried Artemia embryos fulfils the nutritional requirements of post-larvae L. vannamei and promotes a better growth than diets not supplemented with Artemia flakes

Avaliação da ação bioinseticida de SBTI e vicilina de Erythrina velutina em enzimas digestivas e membrana peritrófica de larvas de Plodia interpunctella (Lepidoptera: Pyralidae)

Plodia interpunctella (Indian meal moth) is a cosmopolitan pest that attacks not only a wide range of stored grain as well other food products. Due to its economic importance several researches have focused in a method with ability to control this pest with few or no damage to the environment. The study of digestive enzymes inhibitors, lectins and chitin-binding proteins, has often been proposed as an alternative to reduce insect damage. In this study we report the major classes of digestive enzymes during larval growth in P. Interpunctella, being those proteinases actives at pH 9.5 and optimum temperature of 50 oC to both larvae of the 3rd instar and pre-pupal stage of development. In vitro and zymogram assays presented the effects of several inhibitors, such as SBTI, TLCK and PMSF to intestinal homogenate of 3rd instar larvae of 62%, 92% and 87% of inhibition and In pre-pupal stage of 87%, 62 % and 55% of inhibition, respectively. Zymograms showed inhibition of two low molecular masses protein bands by TLCK and that in presence of SBTI were retarded. These results are indicative of predominance of digestive serine proteinases in gut homogenate from Plodia interpunctella larvae. This serine proteinase was then used as a target to evaluate the effect of SBTI on larvae in in vivo assay. Effect of SBTI on mortality and larval mass was not observed at until 4% of concentration (w/w) in diets. Chitin, another target to insecticidal proteins, was observed by chemical method. Moreover, optic microscopy confirmed the presence of a peritrophic membrane. Established this target, in vivo effect of EvV, a chitin binding vicilin, evaluated during the larval development of P. interpunctella and was obtained a LD50 of 0,23% and WD50 of 0,27% to this protein. Mechanism of action was proposed through of the in vivo digestibility of EvV methodology. During the passage through the larval digestive tract was observed that EvV was susceptible to digestive enzymes and a reactive fragment, visualized by Western blotting, produced by digestion was recovered after dissociation of the peritrophic membrane. The bound of EvV to peritrophic membrane was confirmed by immunohystochemical assays that showed strong immunofluorescent signal of EvV-FITC binding and peritrophic membrane. These results are a indicative that vicilins could be utilized as potential insecticide to Plodia interpunctella and a control methods using EvV as bioinsecticide should be studied to reduce lost caused by storage insect pests

Potencial larvicida de extratos de plantas regionais no controle de larvas de Aedes Aegypti (Diptera:Culicidae)

Dengue, amongst the virus illnesses one can get by vectorial transmission, is the one that causes more impact in the morbidity and mortality of world s population. The resistance to the insecticides has caused difficulties to control of vector insect (Aedes aegypti) and has stimulated a search for vegetables with larvicidal activity. The biodiversity of Caatinga is barely known and it is potential of use even less. Some plants of this biome are commercialized in free fairs northeast of Brazil, because of its phytotherapics properties. The vegetables in this study had been selected by means of a questionnaire applied between grass salesmen and natives of the Serido region from Rio Grande do Norte state; culicids eggs had been acquired with traps and placed in container with water for the larva birth. Thirty larvae had been used in each group (a group control and five experimental groups), with four repetitions four times. The vegetables had been submitted to the processes of decoction, infusion and maceration in the standard concentration of 100g of the vegetable of study in 1l of H2O and analyzed after ½, 1, 2, 4, 8, 12, 24 and 48 hours for verification of the average lethal dose (LD50) from the groups with thirty larva. The LD50 was analyzed in different concentrations (50g/l, 100g/l, 150g/l, 200g/l e 300g/l) of Aspidosperma pyrifolium Mart. 48 extracts of rind, leaf and stem of the seven vegetal species: Aspidosperma pyrifolium Mart., Mimosa verrucosa Benth, Mimosa hostilis (Mart.) Benth., Myracrodruon urundeuva Allemão, Ximenia americana L, Bumelia sartorum Mart Zizyphus joazeiro Mart, had been analyzed. The extracts proceeding from the three methods were submitted to the freezedrying, to evaluate and to quantify substances extracted in each process. The results had shown that Aspidosperma pyrifolium Mart. and Myracrodruon urundeuva Allemão are the species that are more distinguished as larvicidal after 24 hours of experiment, in all used processes of extraction in the assays. The Zizyphus joazeiro Mart species has not shown larvicidal activity in none of the assays. In relation to the extraction method, the decoction was the most efficient method in the mortality tax of the A. aegypti larvae

Tributos indiretos na cadeia produtiva do camarão

The state of Rio Grande do Norte counts with a relevant potential in the shrimp farming supply chain. In the larviculture step the state responds for more than half of the national production. In the farming step it is the second largest producer. In the industrial step, its industries have almost 40% of the shrimp processing capacity of the northeast of Brazil. However, this country has the highest tax rate comparing with the main shrimp producer countries. Considering the influence of taxes in the competition among companies, the main goal of this research is to analyze the impact of indirect taxes in the above steps of the supply chain. To achieve it, it will be used the data of the 2011 Census of the Shrimp Farming and it will be applied the Herfindahl-Hirschman Index to identify the market form of those steps. In order to contribute with the characterization of the supply chain, CEO´s of farms and industries will be interviewed. The price-elasticity of the shrimp larvae, the in natura shrimp and the processed shrimp will be analyzed in order to verify the possibility that each one of those three steps has to pass-through the onus of the end of benefit over the ICMS. The data analysis shows that the larviculture step functions as a duopoly and, facing the end of that benefit, it will be able to pass-through most its onus to the farming step. On the other hand, this step functions similar to a perfect competing market, which diminishes its capacity to pass-through that onus to the processing step. This step operates as oligopoly with a lower concentration than the larviculture step but, due to the fact that it faces an oligopsony, it will end up assuming most of that onus, which will cause a decrease in the amount of processed shrimp. It is concluded that the end of that benefit would impact negatively, in this state, the supply chain at all, but mainly the farming and the industrial steps

Avaliação espaço-temporal do zooplâncton da laguna estuarina de Guaraíras (RN, Brasil)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Purificação, caracterização e atividade bioinseticida de um inibidor de tripsina de sementes de Crotalaria pallida

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulphate fractionation, affinity chromatography on immobilized Trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named ITC, had Mr of 32.5 kDa by SDS-PAGE and was composed by two subunits with 27.7 and 5.6 kDa linked by disulphide bridges, a typical characteristic of Kunitz-Inhibitor family. ITC was stable until 50°C, and at 100°C its residual activity was of about 60%. Also, ITC was stable at pHs 2 to 12. The inhibition of trypsin by ITC was non-competitive, with a Ki of 8,8 x 10-7M. ITC inhibits weakly other serine proteinases such as chymotrypsin and elastase. The inhibition of papain (44% of inhibition), a cysteine proteinase was an indicative of the bi-functionality of ITC. In vitro assays against digestive proteinases from several Lepdoptera, Diptera and Coleoptera pests were made. ITC inhibited in 100% digestive enzymes of Ceratitis capitata (fruit fly), Spodoptera frugiperda and Alabama argillacea, the last one being a cotton pest. It also inhibited in 74.4% Callosobruchus maculatus (bean weevil) digestive enzymes, a Coleoptera pest. ITC, when added in artificial diet models, affected weakly the development of C. capitata larvae and it had a WD50 of 2.65% to C. maculatus larvae

Atividade bioinseticida e mecanismo de ação de vicilinas de sementes Erythrina velutina sobre moscas-das frutas Ceratitis capitata

The fruit fly Ceratitis capitata is considered the most destructive pest of the world fruitculture. Many pest management practices, mainly based on agrochemicals, have been developed to allow the world-wide commerce of fruit. Solutions to decrease the use of synthetic insecticides in agriculture are based on the development of new target-specific compounds which cause less damage to the environment, especially vegetal proteins with insecticidal effects. The aim of this work was to evaluate the deleterious effect of a purified vicilin of E. velutina (EvV) seeds to C. capitata larvae and adult insects and to investigate the mechanisms involved in these effects. EvV was purified, characterized and its deleterious effect was tested in bioassay systems. EvV mechanism of action was determined by immunodetection techniques and fluorescence localization in chitin structures that are present in C. capitata digestory system. EvV is a glycoprotein with affinity to chitin. Its molecular weight, of 216,57 kDa, was determined by gel filtration chromatography in FPLC system. Using SDS-PAGE, it was possible to observe EvV dissociation in two main subunits of 54,8 and 50,8 kDa. When it was submitted to eletrophoresis in native conditions, EvV presented only one band of acid characteristic. The WD50 and LD50 values found in the bioassays were 0,13% and 0,14% (w/w), respectively for the larvae. EvV deleterious effects were related to the binding to chitin structures presented in peritrophic membrane and gut epithelial cells, associated with its low digestibility in C. capitata digestive tract. The results described herein are the first demonstration of the larvicidal effects of plant protein on C. capitata larvae. EvV may be part of the pest management programs, in the toxic bait composition, or an alternative in plant improvement program

Purificação, caracterização e análise da atividade bioinseticida, de um inibidor de tripsina em sementes de catanduva (Piptadenia moniliformis)

One Kunitz-type trypsin inhibitors (PmTI) was purified from Piptadenia moniliformis seeds, a tree of the sub-family Mimosoideae, by TCA precipitation, affinity chromatography on immobilized trypsin-Sepharose, DEAE cellulose (ion exchange) and Superose 12 (molecular exclusion) column FPLC/AKTA. The inhibitor has Mr of 25 kDa by SDS-PAGE and chromatography molecular exclusion. The N-terminal sequence of this inhibitor showed high homology with other family Kunitz inhibitors. This also stable variations in temperature and pH and showed a small decrease in its activity when incubated with DDT in the concentration of 100mM for 120 minutes. The inhibition of trypsin by PmTI was competitive, with Ki of 1.57 x10-11 M. The activity of trypsin was effectively inhibited by percentage of inhibition of 100%, among enzymes tested, was not detected inhibition for the bromelain, was weak inhibitor of pancreatic elastase (3.17% of inhibition) and inhibited by 76.42% elastase of neutrophils, and inhibited in a moderate, chymotrypsin and papain with percentage of inhibition of 42.96% and 23.10% respectively. In vitro assays against digestive proteinases from Lepidoptera, Diptera and Coleoptera pests were carried out. Several degrees of inhibition were found. For Anthonomus grandis and Ceratitis capitata the inhibition was 89.93% and 70.52%, respectively, and the enzymes of Zabrotes subfasciatus and Callosobruchus maculatus were inhibited by 5.96% and 9.41%, respectively, and the enzymes of Plodia. interpunctella and Castnia licus were inhibited by 59.94% and 23.67, respectively. In vivo assays, was observed reduction in the development of larvae in 4rd instar of C. capitata, when PmTI was added to the artificial diet, getting WD50 and LD50 of 0.30% and 0.33%, respectively. These results suggest that this inhibitor could be a strong candidate to plant management programs cross transgenic

Determinação de motivos de ligação à quitina em vicilinas de Canavalia ensiformis e Vigna unguiculata através de métodos in silico e relação com suas toxicidades para o bruquídeo Callosobruchus maculatus (Coleoptera:Bruchidae)

Chitin is an important structural component of the cellular wall of fungi and exoskeleton of many invertebrate plagues, such as insects and nematodes. In digestory systems of insects it forms a named matrix of peritrophic membrane. One of the most studied interaction models protein-carbohydrate is the model that involves chitin-binding proteins. Among the involved characterized domains already in this interaction if they detach the hevein domain (HD), from of Hevea brasiliensis (Rubber tree), the R&R consensus domain (R&R), found in cuticular proteins of insects, and the motif called in this study as conglicinin motif (CD), found in the cristallography structure of the β-conglicinin bounded with GlcNac. These three chitin-binding domains had been used to determine which of them could be involved in silico in the interaction of Canavalia ensiformis and Vigna unguiculata vicilins with chitin, as well as associate these results with the WD50 of these vicilins for Callosobruchus maculatus larvae. The technique of comparative modeling was used for construction of the model 3D of the vicilin of V. unguiculata, that was not found in the data bases. Using the ClustalW program it was gotten localization of these domains in the vicilins primary structure. The domains R&R and CD had been found with bigger homology in the vicilins primary sequences and had been target of interaction studies. Through program GRAMM models of interaction ( dockings ) of the vicilins with GlcNac had been gotten. The results had shown that, through analysis in silico, HD is not part of the vicilins structures, proving the result gotten with the alignment of the primary sequences; the R&R domain, although not to have structural similarity in the vicilins, probably it has a participation in the activity of interaction of these with GlcNac; whereas the CD domain participates directly in the interaction of the vicilins with GlcNac. These results in silico show that the amino acid number, the types and the amount of binding made for the CD motif with GlcNac seem to be directly associates to the deleterious power that these vicilins show for C. maculatus larvae. This can give an initial step in the briefing of as the vicilins interact with alive chitin in and exert its toxic power for insects that possess peritrophic membrane

Isolamento de uma quitinase extraída do cefalotórax do camarão marinho Litopenaeus schmitti (BURKENROAD, 1936) : avaliação de suas atividades antimicrobiana e larvicida

Chitinases are enzymes involved in degradation of chitin and are present in a range of organisms, including those that do not contain chitin, such as bacteria, viruses, plants and animals, and play important physiological and ecological roles. Chitin is hydrolyzed by a chitinolytic system classified as: endo-chitinases, exo-chitinases and N-acetyl-b-D-glucosaminidases. In this study a Litochitinase1 extracted from the cephalotorax of the shrimp Litopenaeus Schmitt was purified 987.32 times using ionexchange chromatography DEAE-Biogel and molecular exclusion Sephacryl S-200. These enzyme presented a molecular mass of about 28.5 kDa. The results, after kinetic assay with the Litochitinase1 using as substrate p-nitrophenyl-N-acetyl-b-Dglucosaminideo, showed apparent Km of 0.51 mM, optimal activity at pH ranging from 5.0 to 6.0, optimum temperature at 55°C and stability when pre-incubated at temperatures of 25, 37, 45, 50 and 55°C. The enzyme showed a range of stability at pH 4.0 to 5.5. HgCl2 inhibited Litochitinase1 while MgCl2 enhances its activity. Antimicrobial tests showed that Litochitinase1 present activity against gram-negative bacterium Escherichia coli in the 800 μg/mL concentration. The larvicidal activity against Aedes aegypti was investigated using crude extracts, F-III (50-80%) and Litochitinase1 at 24 and 48 hours. The results showed larvicidal activity in all these samples with EC50 values of 6.59 mg/mL for crude extract, 5.36 mg/mL for F-III and 0.71 mg/mL for Litochitinase1 at 24 hours and 3.22 and 0.49 mg/mL for the F-III and Litochitinase1 at 48 hours, respectively. Other experiments confirmed the presence of chitin in the midgut of Aedes aegypti larvae, which may be suffering the action of Litochitinase1 killing the larvae, but also the absence of contaminating proteins as serine proteinase inhibitors and lectins in the crude extract, F-III and Litochitinase1, indicating that the death of the larvae is by action of the Litochitinase1. We also observed that the enzymes extracted from intestinal homogenate of the larvae no have activity on Litochitinase1. These results indicate that the enzyme can be used as an alternative to control of infections caused by Escherichia coli and reducing the infestation of the mosquito vector of dengue.