Estudo do genoma do vírus causador da mionecrose infecciosa em camarões e desenvolvimento de métodos para detecção de diferentes variantes

Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites

Processo biotecnológico voltado a produção de estoques poliploides do camarão-branco litopenaeus vannamei (decapoda, penaeidae)

without practical results so far. Protocols used in biotechnological cultured aquatic organisms aimed at increasing growth rates and disease resistance, have been studied and perfected. Among the available techniques, the application of chromosomal manipulation, although still nascent, is presented as a tool aimed at mitigating ecological and economical issues in shrimp farming. The polyploidization artificial method already employed in fish and shellfish, has been widely researched for use in farmed shrimp. Some limitations of this method of expansion in shrimp refer to a better knowledge of cytogenetic aspects, the level of sexual dimorphism and performance in growing conditions. To contribute on some of these issues, the present study aimed to characterize cytogenetic species Litopenaeus vannamei (Decapoda) and Artemia franciscana (Anostraca), analyze the effectiveness of methods for detection of ploidy, through the use of flow cytometry in processes of induction polyploidy cold thermal shock at different stages of development of newly fertilized eggs. Additionally, aimed also the qualitative and quantitative comparison of larval development between diploid and polyploid organisms, besides the identification of sexual dimorphism in L. vannamei, through geometric morphometrics. The results provide information relevant to the improvement and widespread use of biotechnological methods applied toward national productivity in shrimp farming

Avaliação ecológico-econômica do manguezal de Macau/RN e a importância da aplicação de práticas preservacionistas pela indústria petrolífera local

This work presents the results of a survey in oil-producing region of the Macau City, northern coast of Rio Grande do Norte. All work was performed under the Project for Monitoring Environmental Change and the Influence of Hydrodynamic forcing on Morphology Beach Grass Fields, Serra Potiguar in Macau, with the support of the Laboratory of Geoprocessing, linked to PRH22 - Training Program in Geology Geophysics and Information Technology Oil and Gas - Department of Geology/CCET/UFRN and the Post-Graduation in Science and Engineering Oil/PPGCEP/UFRN. Within the economic-ecological context, this paper assesses the importance of mangrove ecosystem in the region of Macau and its surroundings as well as in the following investigative exploration of potential areas for projects involving reforestation and / or Environmental Restoration. At first it was confirmed the ecological potential of mangrove forests, with primary functions: (i) protection and stabilization of the shoreline, (ii) nursery of marine life, and (iii) source of organic matter to aquatic ecosystems, (iv) refuge of species, among others. In the second phase, using Landsat imagery and techniques of Digital Image Processing (DIP), I came across about 18,000 acres of land that can be worked on environmental projects, being inserted in the rules signed the Kyoto Protocol to the market carbon. The results also revealed a total area of 14,723.75 hectares of activity of shrimp production and salting that can be harnessed for the social, economic and environmental potential of the region, considering that over 60% of this area, ie, 8,800 acres, may be used in the planting of the genus Avicennia considered by the literature that the species best sequesters atmospheric carbon, reaching a mean value of 59.79 tons / ha of mangrove

Influência de fatores ambientais na incidência do vírus da infecção hipodermal e necrose hematopoiética (IHHNV) no camarão Litopenaeus vannamei cultivado em fazendas do Estado do Rio Grande do Norte (RN)

The shrimp farming industry is the most profitable area of the aquaculture at Rio Grande do Norte (RN) state, which is one of the largest producers in Brazil. However the infections that affect the shrimp cause major economic losses. The infection is a result of the interaction between the shrimp, the environment and pathogen. The change of these factors may lead to a condition of stress and susceptibility to opportunistic infections. One of these infections caused by Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV) is widely distributed in several countries and affects a wide range of hosts. To optimize conditions for production of Litopenaeus vannamei shrimp, the more species cultivated in Brazil, it is necessary to understand the effects of environmental factors in the susceptibility of this species to infections. The aim of this study was to determine the IHHNV prevalence and to investigate the influence of environmental factors as salinity, temperature, stocking density, dissolved oxygen and rainfall in the IHHNV incidence in L. vannamei grown in farms, in the RN state. To determine the IHHNV prevalence were used 1089 samples of L. vannamei collected in seven farms. To perform the study about the influence of environmental factors, 525 samples of L. vannamei shrimp were collected in eight farms located in regions of low (0-1 ), medium (21-30 ) and high (38-57 ) salinity, using extensive (≤15 shrimp/m2 ), semi-intensive (18-33 shrimp/m2) or intensive (>36 shrimp/m2) stocking density systems. The IHHNV infection was determined in pleopod and hemolymph using the polymerase chain reaction (PCR). The environmental factors were recorded during the collection of animals, using a refractometer to measure the salinity and a multi-parameter meter to measure the temperature and concentration of dissolved oxygen in the water. The IHHNV prevalence in RN was 43% (468 infected shrimp out of 1089), varying on different farms. On the seven farms studied, IHHNV prevalence ranged from 18.6% to 54.8%. The infection rates in the shrimp cultured in low, medium and high salinity were respectively 43.10% (125/290), 31.2% (15/48) and 24.6% (46/187) and was significantly higher in shrimp grown in low salinity (P<0.001). The infection rates in ponds of extensive, semi-intensive and intensive systems were respectively, 28.7%, 28.28% and 47.84%, and was significantly higher in high stocking densities (P<0.001). This study indicated a high IHHNV prevalence and a significant effect of salinity and stocking density, but not of the temperature, rainfall and dissolved oxygen on the IHHNV infection rate in the L. vannamei shrimp cultured in the northeastern Brazil

Efeito da salinidade, densidade de estocagem e da infecção hipodermal e necrose hematopoiética (IHHN) na imunidade do camarão Litopenaeus vannamei cultivados em fazendas do Rio Grande do Norte

The main problem faced by the shrimp industry are the infectious diseases. The hypodermal and hematopoietic necrosis infection (IHHN) is one of the major cause of disease in the cultured shrimp, Litopenaeus vannamei. Environmental changes involving water quality, oxygen concentration, salinity, temperature, stocking density, presence of pathogens, among others, triggering a stressing condition for the cultured shrimp, weakening them and allowing the outbreak of diseases. The stress on the animal leads to a change in the molecules immune response components, which can be used as indicators of shrimp health. Thus, the objective of the present study was to evaluate the effect of salinity, stocking density and IHHNV infection on the L. vannamei shrimp. The immune parameters used to check the shrimp health were the total hemocytes counts (THC), the agglutinating activity (AA) and the clotting time (CT) of the serum of shrimp. These parameters were analyzed in healthy and IHHNV-infected shrimp, grown in low (0-0.5 ), medium (19-24 ) and high (> 38 ) salinity, and extensive (7-12 cam.m-2), semi-intensive (15-25 cam.m-2) and intensive (33-45 cam.m -2) stocking density. The IHHNV infection rate was significantly higher in low salinity (P<0.005) and intensive density (P<0.005), both stressful conditions for L. vannamei. Low salinity significantly increased THC (P<0.05) and decreased and CT (P<0.05) in healthy and infected shrimp, but AA (P<0.05) significantly decreased in healthy shrimp at medium salinity. Culture intensification did not affect the THC, AA and CT of healthy and infected shrimp (P>0.05). The IHHNV infection did not affect any immune parameters of shrimp cultured at different salinities and stocking densities. It is necessary to emphasize that this study was conducted in shrimp grown in ponds, where several environmental factors are acting simultaneously. Thus, further studies are needed about the influence of other environmental factors on the immune parameters of shrimp cultured in pond

Neustonic distribution of decapod planktonic stages and competence of brachyuran megalopae in coastal waters

Settlement rate may not reflect larval supply to coastal waters in different marine invertebrates and demersal fishes. The importance of near-shore oceanography and behaviour of late larval stages may be underestimated. The present study conducted neustonic sampling over station grids and along full-length transects at two embayments in south-eastern Brazil to (1) compare diurnal and nocturnal occurrence of most frequent decapod stages to assess their vertical movements, (2) describe the formation of larval patches and (3) measure competence of crab megalopae according to their distance to recruitment grounds. Several shrimp species apparently undergo a diel vertical migration, swimming crab megalopae showed no vertical movements and megalopae of the intertidal crab Pachygrapsus transversus revealed a reversed vertical migration. During the day, crab megalopae aggregated in convergence zones just below surface slicks. These larvae consisted of advanced, pre-moult stages, at both mid-bay and near-shore patches. Competence, measured as the time to metamorphosis in captivity, was similar between larval patches within each taxon. Yet, subtidal portunids moulted faster to juveniles than intertidal grapsids, possibly because they were closer to settlement grounds. Megalopae of Pachygrapsus from benthic collectors moulted faster than those from bay areas. These results suggest that alternative vertical migration patterns of late megalopae favour onshore transport, and actual competence takes place very close to suitable substrates, where larvae may remain for days before settlement. Lack of correlation between larval supply and settlement for Pachygrapsus suggests that biological processes, besides onshore transport, may play an important role in determining settlement success of coastal crabs.

Camarão Litopenaeus vannamei: componentes de importância nutricional na carne e nos resíduos do beneficiamento

Farming of marine shrimp is growing worldwide and the Litopenaeus vannamei (L. vannamei) shrimp is the species most widely cultivated. Shrimp is an attractive food for its nutritional value and sensory aspects, being essential the maintenance of this attributes throughout storage, which takes place largely under freezing. The aim of this research was to evaluate quality characteristics of Litopenaeus vannamei shrimp, during freezing storage and to verify the effect of rosemary (Rosmarinus officinalis) adding. Considering the reutilization of processing shrimp wastes, total carotenoids analysis were conducted in waste of Litopenaeus vannamei shrimp and in the flour obtained after dryer. Monthly physicochemical and sensorial analysis were carried out on shrimp stored at 28,3 ± 3,8ºC for 180 days. Samples were placed in polyethylene bags and were categorized as whole shrimp (WS), peeled shrimp (PS), and PS with 0,5% dehydrated rosemary (RS). TBARS, pH, total carotenoid and sensorial Quantitative Descriptive Analysis (QDA) were carried out. Carotenoid total analysis was conducted in fresh wastes and processed flour (0 day) and after 60, 120 and 180 days of frozen storage. After 180 days, RS had lower pH (p = 0.001) and TBARS (p = 0.001) values and higher carotenoids (p = 0.003), while WS showed higher carotenoid losses. Sensory analysis showed that WS were firmer although rancid taste and smell were perceived with greater intensity (p = 0.001). Rancid taste was detected in RS only at 120 days at significantly lower intensity (p = 0.001) than WS and PS. Fresh wastes had 42.74μg/g of total carotenoids and processed flour 98.51μg/g. After 180 days of frozen storage, total carotenoids were significantly lower than 0 day (p<0,05). The addition of rosemary can improve sensory quality of frozen shrimp and reduce nutritional losses during storage. Shrimp wastes and flour of L. vannamei shrimp showed considerable astaxanthin content however, during storage it was observed losses in this pigment

Biological features of a puzzling symbiotic association between the hermit crab Dardanus insignis and the porcellanid crab Porcellana Sayana (Crustacea)

The symbiotic lifestyle is widespread among porcellanid crabs, which maintain ecological and co-evolutionary associations with annelid polychaetes, poriferans, cnidarians, echinoderms, gastropod. mollusks, and other crustaceans such as shrimps and hermit crabs, among others. We investigated the ecological association between the hermit crab Dardanus insignis and the porcellanid Porcellana sayana, in southeastern Brazil. Porcellanid crabs, hermit crabs, and available shells were collected monthly from July 2001 to June 2003, with a shrimp boat equipped with two double-rig trawl nets. The majority of P. sayana specimens were collected in shells occupied by D. insignis (96.6%); a few were found in empty shells (3.4%). The catch of both symbionts and hosts increased with increasing depth, with the highest occurrence at 35 m. The F. sayana crabs of various sizes could be found solitary or forming aggregations of up to 14 individuals per host, showing no sex or size segregation. In spite of the high diversity of shell species occupied by the hermit crabs and also available in the field, only a few of them were also utilized by P. sayana. The majority (93%) of shells utilized by P. sayana also hosted other symbiont species, constituting the basis of extensive symbiotic complexes. Thus, the ecological relationship between D. insignis and P. sayana may be classified as a non-obligate and non-specific symbiosis that may also involve other facultative organisms such as sea anemones. (C) 2008 Elsevier B.V. All rights reserved.

Intra- and extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a phylogenetic perspective on the conquest of fresh water by the decapod Crustacea

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

Qualidade da água e composição fitoplanctônica de um viveiro de piscicultura utilizado para fins de pesca esportiva e irrigação

Water quality of a fish pond used by UNESP, at Jaboticabal, SP, Brazil, for both fishing and irrigation has been evaluated according to certain limnological parameters and to its phytoplankton composition. During the year, four sampling sites were assigned at the pond: P(1)=inlet water in the pond; P(2)=inlet water from a shrimp pond; P(3)=irrigation spot; and P(4)=outlet water (water flowing directly into another pond). The collected data show a water pond with high concentration of nutrients and chlorophyll-a, and high values of electrical conductivity, total hardness and total dissolved solids, proceeding from the above pond. These factors affected substantially some biological parameters, such as chlorophyll-a and phytoplankton. In P(3), whereas class Zygnematophyceae showed the highest specimens richness, with 34.01%, the Chlorophyceae, that had the highest number of genera, comprehended 33.52% of the total number of individuals. The use of the water fish pond for irrigation must be re-evaluated, due to eutrophic conditions of the water, the presence of potentially toxic cyanobacteria, so as the proper management employed.

Characterization of the genetic variability among Caesalpinia pulcherrima (L.) Sw. (Fabaceae) plants using RAPD molecular markers

O flamboyanzinho (Caesalpinia pulcherrima, Fabaceae) é muito utilizado como cerca-viva e na arborização de ruas, parques e jardins. de florescimento exuberante, suas flores podem apresentar coloração rosa, laranja ou amarela, conforme a variedade. Como características florais são essenciais para a definição do valor comercial de espécies ornamentais, o objetivo principal do trabalho foi verificar a existência de polimorfismo entre plantas de C. pulcherrima, que produzem flores de diferentes colorações, por marcadores moleculares RAPD. Foram escolhidas aleatoriamente 30 plantas adultas, cultivadas no município de Jaboticabal, Estado de São Paulo, para a retirada de amostras foliares para a extração de DNA. Dentre essas matrizes, 20 possuem flores alaranjadas, oito, flores amarelas, e apenas duas produzem flores de coloração rosa. Dos 140 primers de RAPD avaliados, 94 foram capazes de ampliar fragmentos definidos, gerando 246 bandas, das quais se observou 100% de bandas monomórficas, indicando que nenhum dos primers utilizados detectou polimorfismo entre os tratamentos. Concluiu-se que a técnica para detecção de polimorfismo por marcadores RAPD não foi eficiente, e que existe a necessidade de se testarem marcadores moleculares mais específicos. Além disso, a característica morfológica cor de flor é, possivelmente, controlada por vários genes ou pela combinação deles.

Caracterização citogenética em Schlumbergera truncata (Haworth) Moran e Schlumbergera × buckleyi (T. Moore) Tjaden (Cactaceae)

O número cromossômico diplóide de Schlumbergera truncata e Schlumbergera x buckleyi, de indivíduos com diferentes tipos de coloração de pétalas, foi determinado usando-se pontas de raízes. A utilização de 8-hidroxiquinoleína 0,003 M à 36 °C por 3 horas possibilitou melhor separação cromossômica. Técnica de bandeamento C e de coloração Giemsa permitiram o estudo cariológico dessas espécies. O híbrido Schlumbergera × buckleyi (rósea) apresenta 2n = 22 cromossomos com fórmula cariotípica 16 M + 6 SM. Schlumbergera truncata, apresentando pétalas nas cores vermelha, branca e pink, possui 2n = 22 cromossomos, formulação cariotípica idêntica à de Schlumbergera × buckleyi, enquanto a planta com flores de coloração amarelada mostrou 2n = 34 cromossomos. A classificação cromossômica foi baseada no índice centromérico. Nas plantas que apresentam coloração vermelha, branca, pink e rósea nas pétalas, o melhor período de obtenção de metáfases corresponde ao período de florescimento. Schlumbergera truncata com flores amareladas apresenta dois picos anuais de divisão mitótica. Esses resultados dão suporte à um melhor entendimento da biologia no gênero Schlumbergera e auxiliam na classificação taxonômica nos casos onde apenas as características fenotípicas não são suficientemente confiáveis para a classificação das plantas no mesmo táxon.

Dimensionamento e sobreposição de nichos dos portunídeos (Decapoda, Brachyura), na Enseada da Fortaleza, Ubatuba, São Paulo, Brasil

The niche breadth of the Portunidae and their overlap on the subtidal sediments of Fortaleza bay, Ubatuba (São Paulo) was analyzed. Samples were made monthly from November/1988 to October/1989, inseven areas of the bay using a shrimp fishery boat equipped with two otter-trawls. Each area was characterized based on environmental factors such as depth, temperature, salinity, dissolved oxygen, organic matter and granulometric composition of the sediment. The number of individuais of each species was registered to each area (resource). Levins's standardized measure (BA) and niche percentage were calculated. Five species of swimming crabs were recorded in this study: Callinectes ornatus Ordway, 1863, Arenaeus cribrarius (Lamarck, 1818), Portunus spinimanus Latreille, 1819, Callinectes danae Smith, 1869 and Portunus spinicarpus (Stimpson, 1871). The widest ecological niche occurred to C. ornatus, present in all sampled areas (generalist species). Such fact can be related to high sediment tolerance when it is compared to P. spinimanus (especialist species) which was limited to the areas with coarse granulometric fractions. Highest niche overlap was verified between C. danae and A. cribrarius may be due to greater salinity tolerance of these species. The low occurrence of P. spinicarpus and its reduced niche size in Fortaleza Bay are due to association of this species to cold water currents (ACAS) more evidente in smaler depths during summer months. One future evaluation of the portunid diet can be useful to complement informations about this important aspect of the marine ecology.

Retardadores de crescimento na produção de plantas floridas envasadas de açafrão-da-cochinchina

A introdução de novos produtos na floricultura brasileira é importante para o crescimento do setor. O açafrão-da-cochinchina (Curcuma alismatifolia) possui inflorescência e folhagem altamente decorativas e grande potencial para produção como florífera envasada, com o auxílio de retardadores de crescimento, usados na produção de plantas compactas e atrativas. Avaliou-se o efeito de retardadores de crescimento na produção de açafrão-da-cochinchina `Pink', de 3/12/2004 a 28/4/2005, sob ambiente protegido, em Holambra (SP). Instalou-se experimento em blocos casualizados com três repetições e onze tratamentos: controle, paclobutrazol (20; 25; 30 e 35 mg i.a./vaso, via substrato), duas, três e cinco pulverizações de daminozide na concentração de 2,125 g L-1 e duas, três e cinco pulverizações de solução de daminozide (1,275 g L-1) mais chlormequat (0,2 g L-1). O propágulo foi plantado em vaso plástico (1,0 L) com substrato comercial e fertirrigado diariamente. Paclobutrazol (35 mg i.a./vaso) reduziu significativamente a altura da parte aérea e da folhagem e o comprimento da haste floral, sem alterar o comprimento da inflorescência e atrasar o ciclo de produção. Entretanto, as plantas não ficaram suficientemente compactas para atender às exigências de qualidade do mercado. Os demais retardadores estudados não afetaram significativamente a altura da folhagem e o comprimento da haste floral comparado ao controle.

Modelos para estimativa da área foliar de Curcuma alismatifolia e Vurcuma zedoaria

O presente estudo determina modelos para estimativa da área foliar de Curcuma alismatifolia e de Curcuma zedoaria. Para utilização destas espécies como ornamentais, é necessário o estabelecimento de técnicas de produção adequadas. Assim, a determinação da área foliar é importante, pois é usada para avaliar a resposta da planta a fatores ambientais e técnicas culturais. O uso de modelos para estimar a área foliar é um método simples, de boa precisão e não destrutivo. No estádio de floração foram coletadas cem folhas de C.alismatifolia ('Pink' e 'White') e de C.zedoaria. Determinaram-se o comprimento (C) e a largura (L) máximos e a área foliar real (AFR), com auxílio de integrador de área foliar (LI-3100). Estudaram-se as relações entre a AFR e o C, L e CL (produto do comprimento pela largura da folha), por meio de modelos de regressão linear. Os modelos AFR = 0,59048 CL (C.alismatifolia 'Pink'), AFR = 6,08410 + 0,52162 CL (C.alismatifolia 'White') e AFR = 0,70233 CL (C.zedoaria) são estatisticamente adequados para estimar a área foliar real.