Formação de biofilmes por Enterococcus faecium sensível e multirresistente aos antimicrobianos: características à proteômica

Enterococcus faecium tem se destacado no cenário das infecções hospitalares, particularmente, amostras portadoras de características de multirresistência. Apesar de serem responsabilizados como agentes etiológicos em diferentes quadros clínicos, fatores associados a patogênese das infecções ainda não estão esclarecidos. Entretanto, sabe-se que a capacidade de formação de biofilmes pode ser responsabilizada como um dos atributos capazes de promover o papel patogênico desses microrganismos. Assim sendo, este estudo se propôs a investigar a capacidade de formação de biofilmes por duas amostras de E. faecium: SS-1274 (derivada da amostra tipo da espécie) e CL-6729 (multirresistente e pertencente a um complexo clonal globalmente disperso). As amostras foram caracterizadas fenotipica e genotipicamente para confirmação da identificação, determinação da susceptibilidade a 17 antimicrobianos, caracterização da concentração mínima inibitória para ampicilina, gentamicina e vancomicina, determinação do genótipo vanA e detecção de genes associados a expressão dos genes asa1, cylA, esp, gelE e hyl. A análise quantitativa da formação por 24h e 72h dos biofilmes foi realizada por metodologia do cristal violeta, em placas de microtitulação de poliestireno. Foram construídas curvas de formação pela avaliação da DO570nm das preparações coradas por cristal violeta em períodos de tempo de 2h a 74h. A influência de subCMIs de ampicilina, gentamicina e vancomicina na formação de biofilmes de E. faecium foi também caracterizada em ensaios de quantificação da biomassa. A presença de DNA e proteínas foi avaliada em ensaios de destacamento e por fluorimetria. A arquitetura, distribuição espacial e reação aos fluorocromos Syto9 e SYPRO Ruby Protein foram evidenciadas por microscopia confocal de varredura a laser. Análises proteômicas através da avaliação por SDS-PAGE e por ESI-Q-TOF também foram empregadas para avaliação de biofilmes versus crescimento planctônico. Nossos resultados demonstraram que a amostra CL-6729 apresentou uma maior biomassa nos tempos analisados. Apesar da menor quantidade de biomassa da amostra SS-1274, o perfil da curva de formação foi semelhante a CL-6729. As análises da matriz por ensaios quantitativos e microscopia confocal revelaram que proteína parece ser um importante constituinte para ambas as amostras, entretanto biofilmes formados na presença de da CMI e durante 24h, parecem sofrer alterações na constituição. Os resultados relativos às espectrometrias de massa sugerem que células de biofilmes de E. faecium podem também estar metabolicamente ativas, devido a identificação de um considerável número de proteínas relacionadas ao metabolismo e divisão celular, similarmente às células planctônicas. No entanto, investigações complementares são necessárias para quantificar as diferenças relacionadas a sua expressão. Foi observado que ambas as amostras independente das variações fenotípicas e genotípicas são capazes de formar biofilmes maduros exibindo constituição e arquitetura similares. Entretanto, nossos resultados sugeriram que cada amostra responde as diferentes situações de acordo com seus determinantes de virulência e resistência.

Caribbean corals in crisis: record thermal stress, bleaching, and mortality in 2005

Background: The rising temperature of the world’s oceans has become a major threat to coral reefs globally as the severity and frequency of mass coral bleaching and mortality events increase. In 2005, high ocean temperatures in the tropical Atlantic and Caribbean resulted in the most severe bleaching event ever recorded in the basin. Methodology/Principal Findings: Satellite-based tools provided warnings for coral reef managers and scientists, guiding both the timing and location of researchers’ field observations as anomalously warm conditions developed and spread across the greater Caribbean region from June to October 2005. Field surveys of bleaching and mortality exceeded prior efforts in detail and extent, and provided a new standard for documenting the effects of bleaching and for testing nowcast and forecast products. Collaborators from 22 countries undertook the most comprehensive documentation of basin-scale bleaching to date and found that over 80% of corals bleached and over 40% died at many sites. The most severe bleaching coincided with waters nearest a western Atlantic warm pool that was centered off the northern end of the Lesser Antilles. Conclusions/Significance: Thermal stress during the 2005 event exceeded any observed from the Caribbean in the prior 20 years, and regionally-averaged temperatures were the warmest in over 150 years. Comparison of satellite data against field surveys demonstrated a significant predictive relationship between accumulated heat stress (measured using NOAA Coral Reef Watch’s Degree Heating Weeks) and bleaching intensity. This severe, widespread bleaching and mortality will undoubtedly have long-term consequences for reef ecosystems and suggests a troubled future for tropical marine ecosystems under a warming climate

Formação de Formadores e Educação Superior na cibercultura: itinerâncias de Grupos de Pesquisa no Facebook

Esta tese origina-se da pesquisa de doutoramento na Universidade do Estado do Rio de Janeiro UERJ - Proped e tem como objetivo compreender a formação do formador no contexto da cibercultura e como este vem estruturando sua atuação na docência universitária. Este trabalho faz parte do eixo 2 da Pesquisa Institucional A cibercultura na era das redes sociais e da mobilidade: novas potencialidades para a formação de professores. O campo da pesquisa foi a rede social Facebook. Constituíram-se praticantes da pesquisa seis professores-formadores do ProPEd e seus orientandos. A pertinência desta pesquisa inscreveu-se num amplo movimento da perspectiva epistemológica da multirreferencialidade com os cotidianos e pelo método atualizado da pesquisa-formação. A intenção primeira deste trabalho consistiu em mapear as experiências e itinerâncias do formador em suas práticas cotidianas nos espaçostempos da cibercultura. Tencionamos que a sistematização de ambiências formativas articulando o espaço da universidade e das redes sociais criou redes de docência e aprendizagem. Como dispositivos de pesquisa, lançamos mão de oficinas, interações nas redes sociais, conversas, participação nos grupos de pesquisa. A tese revela que a formação do formador na cibercultura forja outros espaçostempos de pesquisa acadêmica e de criação de dispositivos online. Revela também que os usos do digital em rede pelos praticantes da pesquisa fizeram emergir processos de orientação coletiva, de experiências formadoras e de articulação da interface cidadeuniversidade, estabelecendo outros sentidos para a prática pedagógica e para a pesquisa acadêmica

Enterococcus faecium: caracterização molecular de amostras isoladas no período anterior e posterior a emergência da resistência à vancomicina no Estado do Rio de Janeiro

Nas últimas décadas, Enterococcus resistentes à vancomicina tem se destacado no cenário hospitalar em todo mundo. A emergência da resistência à vancomicina no Estado do Rio de Janeiro foi registrada em 2000, na espécie Enterococcus faecalis. Em seguida, Enterococcus faecium passou a ser prevalente. O objetivo deste estudo foi avaliar a diversidade genética das amostras de E. faecium sensíveis (VSEfm) e resistentes (VREfm) à vancomicina, isoladas em diferentes instituições de saúde do Estado do Rio de Janeiro no período de 1993 a 2008. As amostras bacterianas foram avaliadas por metodologia de eletroforese em campo pulsado (PFGE), após restrição com Smal, análise do polimorfismo numérico de segmentos repetitivos (MLVA) e tipagem por sequenciamento de múltiplos loci (MLST); além da detecção de marcadores fenotípicos, como a resistência à ampicilina e à ciprofloxacina, e genotípicos, como determinantes genéticos de virulência, que estão vinculados a um complexo clonal globalmente disperso (CC17). A diversidade de Tn1546 (que alberga o conjunto gênico vanA) foi determinada por amplificação e restrição com ClaI. Um grupo clonal prevalente foi observado pela metodologia de PFGE e agrupou amostras isoladas, principalmente, no período de 2004 a 2006, estando disseminado por diversas instituições de saúde do Estado, indicando transmissão inter- e intra-hospitalar. A avaliação por MLVA identificou dois MTs prevalentes dentre as amostras VREfm: MT12 relacionado à maioria das amostras dos principais grupos clonais identificados por PFGE e, o MT159 que apresentou frequência aumentada no ano de 2008. A análise por MLST destacou o ST78 associado aos principais grupos clonais definidos por PFGE, bem como, ao MT12. Também foi observada correlação entre o ST412 associado ao grupo clonal formado por apenas amostras de 2008 e o MT159. Foi notório por MLST que as amostras VREfm do Estado do Rio de Janeiro, no período do estudo, estiveram relacionadas CC17, juntamente com algumas amostras VSEfm. Entretanto, os principais marcadores de resistência e de virulência, característicos de CC17, estiveram mais relacionados as amostras VREfm do que as VSEfm, como resistência à ampicilina e à ciprofloxacina; e a presença do gene esp e do alelo purK1. A identificação de VSEfm pertencentes ao CC17 sugerem que amostras já adaptadas ao ambiente hospitalar, podem ter adquirdo o determinante de resistência vanA, facilitando a emergência de amostras de VREfm. Adicionalmente, nossos dados sugerem uma modificação no cenário de amostras VREfm no Rio de Janeiro. Pois a dominância de um grupo clonal prevalente (PFGE- X; MT12; ST78) pode estar sendo substituída pela possível emergência de um novo grupo clonal, que foi característico de amostras mais recentes (PFGE-XV; MT159; ST412), apontando assim, para um novo curso na epidemiologia dos E. faecium no Estado. Um perfil de restrição prevalente de Tn1546 idêntico ao protótipo foi observado, apontando que a disseminação da resistência à vancomicina ocorreu por também por transferência horizontal. Entretanto, alterações do tipo deleção e presença de elementos de inserção com aproximadamente 2 kb foram observadas em um número reduzido das amostras. A incongruência no genótipo vanA, em relação ao fenótipo, foi observado para uma amostra. Considera-se fundamental o acompanhamento da disseminação de VREfm, particularmente diante da elevada frequência de amostras pertencentes a um complexo clonal que conjuga multirresistência com virulência, com elevado potencial de adaptabilidade e disseminação.

A exploração e a produção não convencionais de petróleo e gás natural:uma proposta de regulação não convencional

Esta dissertação debruçou-se sobre o estudo da regulação da exploração e da produção de recursos não convencionais de petróleo e gás natural e de controvérsias acerca da matéria. Partimos de fundamentos e segmentos do Direito Internacional Contemporâneo, sua interseção com o direito interno, precisamente no que diz respeito ao tema da regulação e os seus novos contornos, para então adentrar na regulação propriamente dita dos recursos não convencionais. Após ilustrarmos os diferentes tratamentos conferidos à exploração e à produção de recursos não convencionais no direito comparado, destacamos, como estudo de caso, o tratamento conferido no Brasil à questão e as controvérsias decorrentes dos não convencionais. Ao final, apresentamos algumas proposições com vistas a viabilizar tal atividade não apenas no Brasil como em demais países, por meio de uma regulação não convencional, isto é, elaborada de forma compartilhada e global, pautada nas Melhores Práticas da Indústria, nos princípios de direito internacional e na governança global, que respeite as peculiaridades dos recursos não convencionais e que conte com uma maior aceitação pelos diferentes atores envolvidos e/ou afetados por essa atividade exploratória, por meio da promoção do diálogo e do esclarecimento necessário. Acreditamos que dessa forma estar-se-ia por estimular uma era de ouro global ao gás natural, a considerar todas as suas vantagens perante demais fontes energéticas, proporcionando o desenvolvimento dos Estados de forma ambientalmente sustentável, resguardando-se os interesses dos investidores e, principalmente, dos principais beneficiários da política energética que são os indivíduos.

Genetic improvement of freshwater prawn, Macrobrachium rosenbergii (de Man) in India. Phase two: annual report for period January to December 2012

Macrobrachiurn rosenbergii is one of the widely cultured freshwater prawn species globally. India was the third largest producer of this species in 2007 and its aquaculture production rose to 43,000 metric tons (t) in 2005 froin less than 500 t in 1995. However, since then production has been declining and in 2008-09 it was 12,856 t, a reduction of more than 70% compared to 2005. There are several contributing factors to this decline, such as slow growth rate, poor survival, disease outbreaks, increase in cost of production, and availability of low risk alternative fish species. However, there is a consensus that poor seed quality leading to unsatisfactory growth and survival rates in ponds is one of the major reasons. Hence, the development of a systematic selective breeding program aimed at improving growth rate and ensuring high survival rate of this species was deemed a high priority. The Central Institute of Freshwater Aquaculture (CIFA), Bhubaneswar, India in collaboration with the WorldFish Center, Malaysia initiated a selective breeding program for this species in 2007.

Multiview stereo via volumetric Graph-Cuts and occlusion robust photo-consistency.

This paper presents a volumetric formulation for the multi-view stereo problem which is amenable to a computationally tractable global optimisation using Graph-cuts. Our approach is to seek the optimal partitioning of 3D space into two regions labelled as "object" and "empty" under a cost functional consisting of the following two terms: (1) A term that forces the boundary between the two regions to pass through photo-consistent locations and (2) a ballooning term that inflates the "object" region. To take account of the effect of occlusion on the first term we use an occlusion robust photo-consistency metric based on Normalised Cross Correlation, which does not assume any geometric knowledge about the reconstructed object. The globally optimal 3D partitioning can be obtained as the minimum cut solution of a weighted graph.

Temporal trends (2000–2011) and influences on fishery-independent catch rates for loggerhead sea turtles (Caretta caretta) at an important coastal foraging region in the southeastern United States

Seasonal trawling was conducted randomly in coastal (depths of 4.6–17 m) waters from St. Augustine, Florida, (29.9°N) to Winyah Bay, South Carolina (33.1°N), during 2000–03, 2008–09, and 2011 to assess annual trends in the relative abundance of sea turtles. A total of 1262 loggerhead sea turtles (Caretta caretta) were captured in 23% (951) of 4207 sampling events. Capture rates (overall and among prevalent 5-cm size classes) were analyzed through the use of a generalized linear model with log link function for the 4097 events that had complete observations for all 25 model parameters. Final models explained 6.6% (70.1–75.0 cm minimum straight-line carapace length [SCLmin]) to 14.9% (75.1–80.0 cm SCLmin) of deviance in the data set. Sampling year, geographic subregion, and distance from shore were retained as significant terms in all final models, and these terms collectively accounted for 6.2% of overall model deviance (range: 4.5–11.7% of variance among 5-cm size classes). We retained 18 parameters only in a subset of final models: 4 as exclusively significant terms, 5 as a mixture of significant or nonsignificant terms, and 9 as exclusively nonsignificant terms. Four parameters also were dropped completely from all final models. The generalized linear model proved appropriate for monitoring trends for this data set that was laden with zero values for catches and was compiled for a globally protected species. Because we could not account for much model deviance, metrics other than those examined in our study may better explain catch variability and, once elucidated, their inclusion in the generalized linear model should improve model fits.

Marine cage culture and the environment: twenty-first century science informing a sustainable industry

Technological innovation has made it possible to grow marine finfish in the coastal and open ocean. Along with this opportunity comes environmental risk. As a federal agency charged with stewardship of the nation’s marine resources, the National Oceanic and Atmospheric Administration (NOAA) requires tools to evaluate the benefits and risks that aquaculture poses in the marine environment, to implement policies and regulations which safeguard our marine and coastal ecosystems, and to inform production designs and operational procedures compatible with marine stewardship. There is an opportunity to apply the best available science and globally proven best management practices to regulate and guide a sustainable United States (U.S.) marine finfish farming aquaculture industry. There are strong economic incentives to develop this industry, and doing so in an environmentally responsible way is possible if stakeholders, the public and regulatory agencies have a clear understanding of the relative risks to the environment and the feasible solutions to minimize, manage or eliminate those risks. This report spans many of the environmental challenges that marine finfish aquaculture faces. We believe that it will serve as a useful tool to those interested in and responsible for the industry and safeguarding the health, productivity and resilience of our marine ecosystems. This report aims to provide a comprehensive review of some predominant environmental risks that marine fish cage culture aquaculture, as it is currently conducted, poses in the marine environment and designs and practices now in use to address these environmental risks in the U.S. and elsewhere. Today’s finfish aquaculture industry has learned, adapted and improved to lessen or eliminate impacts to the marine habitats in which it operates. What progress has been made? What has been learned? How have practices changed and what are the results in terms of water quality, benthic, and other environmental effects? To answer these questions we conducted a critical review of the large body of scientific work published since 2000 on the environmental impacts of marine finfish aquaculture around the world. Our report includes results, findings and recommendations from over 420 papers, primarily from peer-reviewed professional journals. This report provides a broad overview of the twenty-first century marine finfish aquaculture industry, with a targeted focus on potential impacts to water quality, sediment chemistry, benthic communities, marine life and sensitive habitats. Other environmental issues including fish health, genetic issues, and feed formulation were beyond the scope of this report and are being addressed in other initiatives and reports. Also absent is detailed information about complex computer simulations that are used to model discharge, assimilation and accumulation of nutrient waste from farms. These tools are instrumental for siting and managing farms, and a comparative analysis of these models is underway by NOAA.

MDNR-NOAA trawl standardization study

Long-term living resource monitoring programs are commonly conducted globally to evaluate trends and impacts of environmental change and management actions. For example, the Woods Hole bottom trawl survey has been conducted since 1963 providing critical information on the biology and distribution of finfish and shellfish in the North Atlantic (Despres-Patango et al. 1988). Similarly in the Chesapeake Bay, the Maryland Department of Natural Resources (MDNR) Summer Blue Crab Trawl survey has been conducted continuously since 1977 providing management-relevant information on the abundance of this important commercial and recreational species. A key component of monitoring program design is standardization of methods over time to allow for a continuous, unbiased data set. However, complete standardization is not always possible where multiple vessels, captains, and crews are required to cover large geographic areas (Tyson et al. 2006). Of equal issue is technological advancement of gear which serves to increase capture efficiency or ease of use. Thus, to maintain consistency and facilitate interpretation of reported data in long-term datasets, it is imperative to understand and quantify the impacts of changes in gear and vessels on catch per unit of effort (CPUE). While vessel changes are inevitable due to ageing fleets and other factors, gear changes often reflect a decision to exploit technological advances. A prime example of this is the otter trawl, a common tool for fisheries monitoring and research worldwide. Historically, trawl nets were constructed of natural materials such as cotton and linen. However modern net construction consists of synthetic materials such as polyamide, polyester, polyethylene, and polypropylene (Nielson et. al. 1983). Over the past several decades, polyamide materials which will be referred to as nylon, has been a standard material used in otter trawl construction. These trawls are typically dipped into a latex coating for increased abrasion resistance, a process that is referred to as “green dipped.” More recently, polyethylene netting has become popular among living resource monitoring agencies. Polyethylene netting, commonly known as sapphire netting, consists of braided filaments that form a very durable material more resistant to abrasion than nylon. Additionally, sapphire netting allows for stronger knot strength during construction of the net further increasing the net’s durability and longevity. Also, sapphire absorbs less water with a specific gravity near 0.91 allowing the material to float as compared to nylon with specific gravity of 1.14 (Nielson et. al. 1983). This same property results in a light weight net which is more efficient in deployment, retrieval and fishing of the net, particularly when towing from small vessels. While there are many advantages to the sapphire netting, no comparative efficiency data is available for these two trawl net types. Traditional nylon netting has been used consistently for decades by the MDDNR to generate long term living resource data sets of great value. However, there is much interest in switching to the advanced materials. In addition, recent collaborative efforts between MDNR and NOAA’s Cooperative Oxford Laboratory (NOAA-COL) require using different vessels for trawling in support of joint projects. In order to continue collaborative programs, or change to more innovative netting materials, the influence of these changes must be demonstrated to be negligible or correction factors determined. Thus, the objective of this study was to examine the influence of trawl net type, vessel type, and their interaction on capture efficiency.

Solomon Islands: Western Hub scoping report

Aquatic agricultural systems (AAS) are places where farming and fishing in freshwater and/or coastal ecosystems contribute significantly to household income and food security. Globally, the livelihoods of many poor and vulnerable people are dependent on these systems. In recognition of the importance of AAS, the CGIAR Research Program (CRP) is undertaking a new generation of global agricultural research programs on key issues affecting global food security and rural development. The overall goal of the research program is to improve the well-being of people dependent on these systems. Solomon Islands is one of five priority countries in the AAS program, led by WorldFish. In Solomon Islands, the AAS program operates in the Malaita Hub (Malaita Province) and the Western Hub (Western Province). This program and its scoping activities are summarized in this report.

Solomon Islands: Western Province situation analysis

Aquatic agricultural systems (AAS) are places where farming and fishing in freshwater and/orscoastal ecosystems contribute significantly to household income and food security. Globally, theslivelihoods of many poor and vulnerable people are dependent on these systems. In recognitionsof the importance of AAS, the CGIAR Research Program (CRP) is undertaking a new generationsof global agricultural research programs on key issues affecting global food security and ruralsdevelopment. The overall goal of the research program is to improve the well-being of peoplesdependent on these systems. Solomon Islands is one of five priority countries in the AAS program,sled by WorldFish. In Solomon Islands, the AAS program operates in the Malaita Hub (MalaitasProvince) and the Western Hub (Western Province). This program and its scoping activities aressummarized in this report.

SPEECH RECOGNITION IN VODIS II.

VODIS II, a research system in which recognition is based on the conventional one-pass connected-word algorithm extended in two ways, is described. Syntactic constraints can now be applied directly via context-free-grammar rules, and the algorithm generates a lattice of candidate word matches rather than a single globally optimal sequence. This lattice is then processed by a chart parser and an intelligent dialogue controller to obtain the most plausible interpretations of the input. A key feature of the VODIS II architecture is that the concept of an abstract word model allows the system to be used with different pattern-matching technologies and hardware. The current system implements the word models on a real-time dynamic-time-warping recognizer.

Optimisation of a piezoelectric system for energy harvesting from traffic vibrations

Piezoelectric systems are viewed as a promising approach to energy harvesting from environmental vibrations. The energy harvested from real vibration sources is usually difficult to estimate analytically. Therefore, it is hard to optimise the associated energy harvesting system. This work investigates the optimisation of a piezoelectric cantilever system using a genetic algorithm based approach with numerical simulations. The genetic algorithm globally considers the effects of each parameter to produce an optimal frequency response to scavenge more energy from the real vibrations while the conventional sinusoidal based method can only optimise the resistive load for a given resonant frequency. Experimental acceleration data from the vibrations of a vehicle-excited manhole cover demonstrates that the optimised harvester automatically selects the right frequency and also synchronously optimises the damper and the resistive load. This method shows great potential for optimizing the energy harvesting systems with real vibration data. ©2009 IEEE.

Finite volume distance field solution applied to medial axis transform

Accurate and efficient computation of the nearest wall distance d (or level set) is important for many areas of computational science/engineering. Differential equation-based distance/ level set algorithms, such as the hyperbolic-natured Eikonal equation, have demonstrated valuable computational efficiency. Here, in the context, as an 'auxiliary' equation to the main flow equations, the Eikonal equation is solved efficiently with two different finite volume approaches (the cell vertex and cell-centered). Application of the distance solution is studied for various geometries. Moreover, a procedure using the differential field to obtain the medial axis transform (MAT) for different geometries is presented. The latter provides a skeleton representation of geometric models that has many useful analysis properties. As an alternative approach to the pure geometric methods (e.g. the Voronoi approach), the current d-MAT procedure bypasses many difficulties that are usually encountered by pure geometric methods, especially in three dimensional space. It is also shown that the d-MAT approach provides the potential to sculpt/control the MAT form for specialized solution purposes. Copyright © 2010 by the American Institute of Aeronautics and Astronautics, Inc.