Studien går ut på att jämföra tre olika styrsystem för ventilation, nämligen CAV (Constant Air Ventilation), VAV (Variabel Air Ventilation) och DCV (Demand Controlled Ventilation). Dessa styrsystem ställs mot varandra i tre olika lokaltyper, med syfte att identifiera det mest energieffektiva styrsystemet. Ett syftet med studien är också att upptäcka vid vilka flöden det lönar sig att arbeta med VAV- och DCV-styrsystemen.

Detta arbete är utfört hos Videum AB i Växjö i syfte att visa på skillnader i energiförbrukning mellan deras ventilationssystem samt hur lång tid det skulle ta för en möjlig investering i ett nytt ventilationssystem att återbetala sig. För att uppnå detta utfördes beräkningar på olika faktor som bidrar till den totala energianvändningen hos ventilationssystemet. I denna undersökning behandlas fläktar, golvaggregat och värmeförluster genom ventilation. Den största påverkande faktorn i den totala energiförbrukningen är värmeförluster genom ventilation. Resultatet av vår undersökning visar att det är billigare att använda i VAV-system än ett CAV-system under rådande förhållanden enligt våra beräkningar. En årlig besparing på 29465 kronor per våning vid byte från CAV-system till VAV-system uppnås. Det är värt att byta ventilationssystem även fast att investeringskostnaden inte sparar in sig under dess livstid. Detta på grund av att det blir en mindre årlig kostnad samt att man kommer närmre regeringens krav på minskad energiförbrukning. Avbetalningstiden för VAV-systemet är 33 år per våning.

Submitted by Alessandra Portugal (alessandradf@ioc.fiocruz.br) on 2013-10-01T21:53:22Z No. of bitstreams: 1 Carolina da Paz Zampier.pdf: 2134379 bytes, checksum: d822f378a88e2cf44b02c5ea60f52ea0 (MD5) / Made available in DSpace on 2013-10-01T21:53:22Z (GMT). No. of bitstreams: 1 Carolina da Paz Zampier.pdf: 2134379 bytes, checksum: d822f378a88e2cf44b02c5ea60f52ea0 (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / A caveolina-1 (Cav-1), uma proteína essencial para a formação de cavéolas, apresenta atividade na modulação da sinalização intracelular. Cav-1 é capaz de interagir com diversas proteínas através de seu domínio CSD (caveolin scaffolding domain) e, em geral, essa interação leva à inibição das proteínas associadas. O peptídeo CSD tem sido utilizado como um mimético de Cav-1 em relação à sua capacidade modulatória sobre a atividade de outras proteínas. Recentemente, tem sido mostrado que Cav-1 é capaz de modular a resposta inflamatória em diversos aspectos. Neste trabalho, examinamos o papel de Cav-1 na regulação da síntese de mediadores inflamatórios por macrófagos. O lipopolissacarídeo (LPS) de E.coli, um protótipo de estímulo inflamatório, foi capaz de induzir a expressão de Cav-1 e Cox-2 em macrófagos peritoneais in vitro. Estas proteínas são induzidas em um curso temporal semelhante, sendo detectadas por Western blot a partir de 3h com níveis de expressão crescentes até 18h. Por imunofluorescência, observamos que Cav-1 e Cox-2 apresentam um padrão de expressão mutualmente exclusivo em macrófagos estimulados com LPS. Mostramos por Western blot que a expressão de Cox-2 é induzida por LPS e que o tratamento com CSD leva à inibição da expressão de Cox- 2, mas não de Cox-1. Observamos, também, a redução parcial dos níveis de PGE2 no sobrenadante de macrófagos estimulados com LPS e tratados com CSD. O tratamento com o peptídeo CSD também foi capaz de reduzir os níveis de IL-1, IL- 6, e IL-12 induzidos por LPS. O LPS induz o aumento da expressão e fosforilação de STAT-1. A fosforilação de STAT-1 foi diminuída após o tratamento com CSD, indicando que Cav-1 modula negativamente a ativação de STAT-1. Estudos posteriores são necessários para complementar os dados obtidos até o momento para esclarecer os mecanismos de modulação da síntese de mediadores inflamatórios por Cav-1. Em conclusão, Cav-1 apresenta uma atividade inibitória sobre a expressão de Cox-2 e produção dos mediadores inflamatórios PGE2, IL1, IL-6, e IL-12 em macrófagos estimulados com LPS in vitro. O mecanismo de inibição possivelmente envolve inibição da ativação de STAT-1. / Caveolin-1 (Cav-1), a protein essential for the formation of caveolae, shows activity in the modulation of intracellular signaling. Cav-1 can interact with several proteins by its caveolin scaffolding domain (CSD) and, in general, this interaction leads to inhibition of associated proteins. The peptide CSD has been used as a Cav- 1 mimetic in relation to its capacity on the modulatory activity of other proteins. Recently, it has been shown that Cav-1 can modulate the inflammatory response in several respects. We examined the role of Cav-1 in regulating the synthesis of inflammatory mediators by macrophages. Lipopolysaccharide (LPS) from E. coli, a prototype of inflammatory stimulus, was able to induce the expression of Cav-1 and Cox-2 in peritoneal macrophages in vitro. These proteins are induced in a similar time course, being detected by Western blot at 3 hours with increasing levels of expression up to 18 hours. By immunofluorescence, we observed that Cav-1 and Cox-2 have a mutually exclusive pattern of expression in macrophages stimulated with LPS. Western blot analysis showed that the expression of Cox-2 is induced by LPS and that treatment with CSD leads to inhibition of Cox-2 but not Cox-1. We also observed the partial reduction of PGE2 levels in supernatants of macrophages stimulated with LPS and treated with CSD. Treatment with CSD peptide was also able to reduce the levels of IL1, IL-6 and IL-12 induced by LPS. LPS induces increased expression and phosphorylation of STAT-1. The phosphorylation of STAT- 1 was decreased after treatment with the CSD, indicating that a Cav-1 negatively modulates activation of STAT-1. Further studies are needed to supplement the data obtained so far to clarify the mechanisms of modulation of the synthesis of inflammatory mediators by Cav-1. In conclusion, Cav-1 shows an inhibitory activity on Cox-2 expression and production of the inflammatory mediators PGE2, IL1β, IL-6 and IL-12 in macrophages stimulated with LPS in vitro. The mechanism of inhibition possibly involves inhibition of STAT-1 activation.

Made available in DSpace on 2015-05-14T13:00:18Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4056353 bytes, checksum: ec317eee623ca7f537cb2773d854de24 (MD5) Previous issue date: 2011-02-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Ca2+ is an intracellular messenger that controls a wide range of physiological functions through changes in its cytosolic concentration ([Ca2+]c). The increase in [Ca2+]c is derived of mobilization from intracellular stores or influx through channels, especially voltage-gated (Cav), present on cell surface. According to scientific literature, sea urchins embryogenesis is a process induced exclusively by the release of Ca2+ from the endoplasmic reticulum, and Ca2+ influx is not necessary for this process. However, there are studies in several species of the animal kingdom where Ca2+ influx is crucial for embryogenesis. Therefore, the aim of this study was to evaluate the involvement of Ca2+ influx at fertilization and embryonic development of Echinometra lucunter, a species of sea urchins with wide distribution on Brazilian coast. Thereby, eggs and embryos of E. lucunter were treated with various pharmacological tools and fertilization and embryonic development were monitored. Incubation of gametes in Ca2+ free medium inhibited fertilization and embryo treatment with Ca2+ chelators blocked embryonic development, suggesting that extracellular Ca2+ is essential for both processes. Cav blockers nifedipine, diltiazem and verapamil were also effective in blocking fertilization and embryo development, showing the importance of these channels to embryogenesis of E. lucunter. Inhibitory effect on embryo development is not associated with modulation of ABC superfamily proteins, since embryonic development was not affected, even under inhibition of these proteins. Verapamil inhibitory effect was reversed by prior addition of valinomycin which may be related to a probable increase of [Ca2+]c induced by K+ ionophore. ouabain, a Na+/K+-ATPase blocker that activates the reverse mode of Na+ /Ca2+, also reversed inhibition of development induced by verapamil. The reversal was not observed when compounds were added to embryos after verapamil, suggesting a temporal profile in inhibitory effect of these blockers. Inhibitory effects of verapamil and Ca2+ chelator EGTA were time-dependent, being absent 50 minutes after fertilization, suggesting that Ca2+ influx is seminal only in the first minutes of embryonic development. However, intracellular Ca2+ is essential for embryogenesis, since treatments with BAPTA-AM (chelator of intracellular Ca2+) and chlorpromazine or Trifluoperazine (Ca2+-calmodulin complex blockers) inhibited E. lucunter embryogenesis. Additionally, it was found that the rotundifolone, a plant-derived compound with vasorelaxing activity, attributed to the blockade of Cav, inhibited E. lucunter embryonic development showing an inhibitory profile similar to observed in vascular smooth muscle. Therefore, these results suggest that Ca2+ influx is essential for Echinometra lucunter embryogenesis and certify the embryonic development of this animal as appropriated pharmacological model for the investigation of natural and synthetic products that interferes in Ca2+ cellular dynamics. / O Ca2+ é um mensageiro intracelular que controla uma ampla variedade de funções fisiológicas por meio de alterações na sua concentração citosólica ([Ca2+]c). O aumento na [Ca2+]c é derivado da liberação a partir de estoques intracelulares ou do influxo através dos canais, principalmente os sensíveis à voltagem (Cav), presentes na superfície celular. De acordo com a literatura científica, a embriogênese de ouriços-do-mar é um processo regulado exclusivamente pela liberação de Ca2+ a partir do retículo endoplasmático, sendo o influxo dispensável para esse processo. Entretanto, há relatos em diversas espécies do reino animal onde o influxo de Ca2+ é crucial para a embriogênese. Portanto, o objetivo deste trabalho foi avaliar a participação do influxo de Ca2+ na fertilização e no desenvolvimento embrionário de Echinometra lucunter, uma espécie de ouriço-do-mar com ampla distribuição na costa Brasileira. Dessa forma, óvulos e embriões de E. lucunter foram tratados com diversas ferramentas farmacológicas e a fertilização e o desenvolvimento embrionário monitorados. A incubação dos gametas em meio isento de Ca2+ inibiu a fertilização e o tratamento dos embriões com quelantes de Ca2+ bloqueou o desenvolvimento embrionário, sugerindo que o Ca2+ extracelular é fundamental para ambos os processos. Os bloqueadores de Cav nifedipina, diltiazem e verapamil também foram eficazes no bloqueio da fertilização e do desenvolvimento embrionário, indicando a importância desses canais para a embriogênese de E. lucunter. O efeito inibitório sobre o desenvolvimento embrionário não está associado à modulação de proteínas da superfamília ABC, uma vez que o desenvolvimento embrionário ocorreu de forma normal, mesmo com a inibição dessas proteínas. O efeito inibitório do verapamil foi revertido pela adição prévia de valinomicina e tal fato pode estar relacionado a um provável aumento da [Ca2+]c induzido por este ionóforo de K+. A ouabaína, um bloqueador da Na+/K+-ATPase, capaz de ativar o modo reverso do trocador Na+/Ca2+, também reverteu a inibição do desenvolvimento induzida pelo verapamil. Essa reversão não foi observada quando os compostos foram adicionados aos embriões após o verapamil, sugerindo uma relação temporal no efeito inibitório desses bloqueadores. O efeito inibitório do verapamil e do quelante de Ca2+ EGTA foi dependente de tempo, sendo ausente 50 minutos após a fertilização, sugerindo que o influxo de Ca2+ é um fator determinante apenas nos primeiros minutos do desenvolvimento embrionário. Contudo, o Ca2+ intracelular é indispensável para a embriogênese, uma vez que os tratamentos com o BAPTA-AM, um quelante intracelular de Ca2+, e com trifluoperazina ou clorpromazina, bloqueadores do complexo Ca2+-calmodulina, inibiram a embriogênese de E. lucunter. Adicionalmente, foi verificado que a rotundifolona, um composto de origem vegetal com atividade vaso-relaxante atribuída ao bloqueio dos Cav, inibiu o desenvolvimento embrionário de E. lucunter, obtendo um perfil inibitório similar ao observado em musculatura lisa vascular. Portanto, esses resultados sugerem que o influxo de Ca2+ é essencial para a embriogênese de Echinometra lucunter e legitima o desenvolvimento embrionário desse animal como um excelente modelo farmacológico para a prospecção de produtos naturais e sintéticos bioativos que interferem na dinâmica celular do Ca2+.

Energieffektiviseringar behövs för att klara klimatmålen. Syftet med detta arbete har därför varit att undersöka ventilationen i förskolebyggnaden Rymden i Eskilstuna kommun. Fokus i arbetet har legat på att göra en analys på energibesparingar där variabelt luftflöde, VAV-system och konstant luftflöde, CAV-system jämförs i tre olika scenarier. Metodiken för att lösa arbetets frågeställningar har varit genom informationsinsamling, beräkningsprogram, intervju samt platsbesök. Informationsinsamling har varit i form av vetenskapliga tidskrifter samt arbeten som behandlar det aktuella problemområdet. Olika programvaror har använts, dessa är MagiCAD, IDA ICE, Sektionsdata 4.21 samt Microsoft Excel. För ökad förståelse för den aktuella byggnaden har platsbesök gjorts på förskolebyggnaden samt en intervju med den projektöransvarige för ventilationen i förskolan. Resultatet visar att den mest energieffektiva systemlösningen är en roterande värmeväxlare med VAV-styrning i kombination med en korsströmsvärmeväxlare för köksavdelningen. VAV-styrning med en sådan systemlösning har en total årlig energianvändning på 20 684 kWh, medan CAV-systemet med samma systemlösning använder 30 900 kWh. Ekonomisk analys visar däremot att CAV-systemet med samma systemlösning är mest lönsam. Den totala livscykelkostnaden, LCC ligger på 2 386 857 kr för CAV-systemet vid en kalkyltid på 30 år, i jämförelse med 2 420 117 kr för VAV-systemet. För att VAV-systemet skall vara lönsam, visar känslighetsanalysen att kalkylräntan måste sjunka från 5 % till 2,56 % eller energiprisutvecklingen öka med 2,44 % årligen eller en sänkning av den årliga underhållskostnaden för VAV-styrning med 2 164 kr. Övrig känslighetsanalys visar att vid nederbörd då personer stannar inomhus är det fördel för VAV-systemet, då skillnaden i total LCC-kostnad sjunker med 1 758 kr. Vid 74 % av personnärvaro minskar LCC-skillnaden mellan systemen från 39 240 kr till 26 371 kr, alltså utgör detta även en fördel för VAV-systemet. Slutsatsen som dras för förskolebyggnaden Framtiden är att större energibesparingar kan göras med ett VAV-system, men ett CAV-system är bättre ur en ekonomisk synpunkt. Känslighetsanalysen visar dock att små förändringar behövs för att VAV-systemet skall bli ekonomiskt lönsamt. Andra faktorer som påverkar valet mellan VAV- och CAV-system är exempelvis en minskning av personnärvaron relativt till det dimensionerade. En sådan minskning utgör en fördel för VAV-systemet. Detta på grund av att med ett CAV-system överventileras byggnaden. Även högre specifik fläkteffekt har en stor betydelse i valet, då mer energiåtgång till fläktarna leder till större energibesparing för VAV-system. En högre temperaturverkningsgrad för luftbehandlingsaggregat medför däremot en fördel för CAV-system. / This thesis work has been done in collaboration with Sweco Systems in Eskilstuna, Sweden. The purpose with this work is to investigate the ventilation in a preschool in Eskilstuna. The focus of the work is to analyze the potential energy savings of using a variable air volume system instead of a constant air volume system. An analysis is also made to investigate the economical profitability of three different scenarios. The method used to solve the problem formulation has been through gathering information, in form of scientific journals in the current problem area. Information has also been gathered through an interview and a site visit. Various software has been used in this thesis work for calculations, such as MagiCAD, IDA ICE, Sektionsdata 4.21 and Microsoft Excel. The result shows that the most energy efficient solution is a rotary heat exchanger with variable air volume control in combination with a cross-flow heat exchanger for the kitchen section. However, the most profitable solution from an economical point of view is the same system, but with constant air volume control. Sensitivity analysis shows that for a profitable variable air volume system, either the interest rate has to be lowered from 5 % to 2,56 %, the energy price needs to increase by 2,44 % yearly. Or maintenance cost for the variable air volume system needs to be lowered by 2 164 kr. The conclusion of this thesis work is that energy savings can be done with a variable air volume system. However from an economical point of view, the constant air volume is better than variable air volume for the investigated preschool. The sensitivity analysis shows that small changes are needed to make a variable air volume system profitable. The choice between the systems are influenced by certain factors. A decrease of the attendance than the dimensioned are an advantage for variable air volume systems. This is due to the fact that a constant air volume system ventilates more than needed. A higher specific fan power number are also of great importance in the choice, as more energy consumption by the fans leads to greater energy savings for variable air volume systems.

Att minska energianvändningen är lika mycket ett globalt mål som ett nationellt mål. Sverige har som mål att öka effektiviseringen av energianvändningen med 20 % fram till år 2020. Det gör att nästan alla sektorer påverkas och inte minst bostads- och servicesektorn som står för nästan 40 % av Sveriges energianvändning. Ventilationssystem är en av de saker som kan bidra till energibesparingen. I den här studien har en nybyggd grundskola simulerats avseende energi och komfort där olika ventilationssystem testats för att se vilket system som är mest lönsamt ekonomiskt, energi- och komfortmässigt. Det som skiljer ventilationssystemen åt är de olika typerna av flödesreglering. Två huvudtyper av ventilationsflödesreglering har testats: ”Variable Air Volume” = Luftflödet varierar efter behovet (VAV) och ”Constant Air Volume” = Konstant luftflöde (CAV). Byggnaden, som är välisolerad och har tung stomme, har även simulerats med ökad personbelastning för att se hur det påverkar ventilationssystemet. Metoden som används för att utföra arbetet är en simuleringsstudie av komparativt slag, där datorprogrammet IDA ICE 4.8 har använts för att modellera byggnaden och utföra simuleringarna. Modellen som har simulerats efterliknar delar av grundskolan Stigslundsskolan i Gävle, i både uppbyggnad, ventilation och användningsgrad. Data har samlats in från konsultföretaget Rambolls databaser med information om projekteringen av skolan och genom personlig kommunikation. Simuleringen av ett skolår har utförts och ett schema för personbelastningen har ställts in för varje sal för att efterlikna det schema som eleverna går efter i dagsläget. Resultatet av simuleringarna visade att VAV -ventilationssystemet som efterliknar det projekterade fallet väl täckte ventilationsbehovet, använde mindre energi än CAV-systemet och skapade ett mycket bra termiskt klimat i byggnaden. Resultatet visade också att ökad personbelastning hade stor inverkan på ventilationssystemet, och medförde bland annat att återbetalningstiden för VAV-systemet ökade från 11 till 30 år. / To reduce the energy use is as much a global goal as it is an international goal. The Swedish goal is to increase the efficiency of energy use with 20 % by year 2020. This means that almost all the sectors are affected, that includes the housing and service sector which accounts for almost 40 % of Sweden’s energy use. The ventilation system is one thing that can contribute to saving energy. In this study a new built primary school has been simulated, regarding energy use and thermal comfort, where different ventilation system has been tested to determine which system are the most profitable in economically, energy and comfortably. The ventilation system different in the flow control. Two main types of flow control have been tested: Variable Air Volume (VAV) and Constant Air Volume (CAV). The building, which is well insulated and has a heavy body has also been simulated with increased personal load to see how it affects the ventilation system. The method used to perform the work is a comparative type simulation study. The computer program IDA ICE 4.8 has been used to build the model and do the simulations. The model that has been simulated resemble the primary school Stigslundsskolan in Gävle in both construction, ventilation and degree of use. Data has been collected from the consulting company Ramboll’s database with information on the planning of the school. Personal communication has also been used to collect information. The simulations have been performed for a school year and a schedule for the personal load has been set for each hall to imitate the schedule that the students are following today. The result of the simulations showed that the VAV ventilation system, which resemble the planned case, cover the ventilation requirements, uses less energy than the CAV system and create a very good thermal climate in the building. The result also shows that the increased personal load had a great influence on the VAV ventilation system and increased the repayment period from 11 to 30 years.

Emerging connected and autonomous vehicle technologies (CAV) provide an opportunity to improve highway capacity and reduce adverse impacts of stop-and-go traffic. To realize the potential benefits of CAV technologies, this study provides insightful methodological and managerial tools in microscopic and macroscopic traffic scales. In the macroscopic scale, this dissertation proposes an analytical method to formulate highway capacity for a mixed traffic environment where a portion of vehicles are CAVs and the remaining are human-driven vehicles (HVs). The proposed analytical mixed traffic highway capacity model is based on a Markov chain representation of spatial distribution of heterogeneous and stochastic headways. This model captures not only the full spectrum of CAV market penetration rates but also all possible values of CAV platooning intensities that largely affect the spatial distribution of different headway types. Numerical experiments verify that this analytical model accurately quantifies the corresponding mixed traffic capacity at various settings. This analytical model allows for examination of the impact of different CAV technology scenarios on mixed traffic capacity. We identify sufficient and necessary conditions for the mixed traffic capacity to increase (or decrease) with CAV market penetration rate and platooning intensity. These theoretical results caution scholars not to take CAVs as a sure means of increasing highway capacity for granted but rather to quantitatively analyze the actual headway settings before drawing any qualitative conclusion. In the microscopic scale, this study develops innovative control strategies to smooth highway traffic using CAV technologies. First, it formulates a simplified traffic smoothing model for guiding movements of CAVs on a general one-lane highway segment. The proposed simplified model is able to control the overall smoothness of a platoon of CAVs and approximately optimize traffic performance in terms of fuel efficiency and driving comfort. The elegant theoretical properties for the general objective function and the associated constraints provides an efficient analytical algorithm for solving this problem to the exact optimum. Numerical examples reveal that this exact algorithm has an efficient computational performance and a satisfactory solution quality. This trajectory-based traffic smoothing concept is then extended to develop a joint trajectory and signal optimization problem. This problem simultaneously solves the optimal CAV trajectory function shape and the signal timing plan to minimize travel time delay and fuel consumption. The proposed algorithm simplifies the vehicle trajectory and fuel consumption functions that leads to an efficient optimization model that provides exact solutions. Numerical experiments reveal that this algorithm is applicable to any signalized crossing points including intersections and work-zones. Further, the model is tested with various traffic conditions and roadway geometries. These control approaches are then extended to a mixed traffic environment with HVs, connected vehicles (CVs), and CAVs by proposing a CAV-based speed harmonization algorithm. This algorithm develops an innovative traffic prediction model to estimate the real-time status of downstream traffic using traffic sensor data and information provided by CVs and CAVs. With this prediction, the algorithm controls the upstream CAVs so that they smoothly hedge against the backward deceleration waves and gradually merge into the downstream traffic with a reasonable speed. This model addresses the full spectrum of CV and CAV market penetration rates and various traffic conditions. Numerical experiments are performed to assess the algorithm performance with different traffic conditions and CV and CAV market penetration rates. The results show significant improvements in damping traffic oscillations and reducing fuel consumption.

Syftet med arbetet var att framställa en beräkningsmodell som behärskar att beräkna ett klimatsystems energiförluster samt livscykelanalys, LCC. Samt att använda denna modell vid jämförelse av två befintliga system. Modellen skapades i programmet Excell och använder sig av angiven indata för att beräkna resultat. Den användes för jämförelse av fyra lokaler i VIDEUM ABs byggnader. Två kontor och två föreläsningssalar jämfördes. Den beräknade skillnaden i energiförbrukning kunde i huvudsak härledas till det ena systemets längre drifttid över helgen samt att ett av kontoren drevs med högt konstantflöde hos ventilation även under frånvaro. Huruvida styrning av system sker efter koldioxidkoncentration eller temperatur verkar spela mindre roll för systemets förluster. Dock påverkar onödigt högt ställda flöden samt för långa driftstider energibehovet desto mer. / The thesis focused on compiling a calculation model suitable for calculating an indoor climate system energy demand and life cycle cost, LCC. The model was created in Excell and uses given input data to calculate the results. The model was used to compare four different premises located in buildings owned by VIDEUM AB in Växjö. Two offices and two lecture halls was compared. The calculated differences in energy demand could be derived to longer operating times during weekends for one system. One office had a large constant air flow even during absence which also led to a greater energy demand. Whether the system was regulated by using carbon dioxide concentrations or temperature as indicators on air quality didn’t seem to affect the energy demand significantly. Unnecessary high flow rates and operating times affects the premises energy demand the more.

A presente tese versa sobre estudos realizados visando a identificação de novos agentes virais em frangos comerciais. No primeiro capítulo, a identificação do girovírus aviário tipo 2 (AGV2) é reportada. Um genoma viral de 2383 nt foi amplificado a partir soros de frangos comerciais por amplificação randômica de DNA. A análise da sequência do produto amplificado, revelou que cerca de 40% das sequencias de nucleotídeos apresentavam similaridade com o genoma do vírus da anemia infecciosa das galinhas (CAV). Dado o baixo grau de identidade com o CAV, o genoma identificado justificou a proposição de um novo tipo de vírus, dentro do gênero gyrovírus foi denominado “girovírus aviário tipo 2” (AGV2). O genoma do AGV2 tem organização semelhante a do CAV, com percentagens de similitude de aminoácidos nas regiões codificantes, correspondentes às proteínas virais VP1, VP2 e VP3 do CAV de 38,8%, 40,3% e 32,2%, respectivamente. A fim de analisar a amplitude da disseminação deste agente, foi realizado um estudo para identificar a ocorrência do AGV2 em granjas de frangos de outras regiões. Para atingir esse objetivo, uma PCR específica ao AGV2 foi desenvolvida para amplificar o DNA viral extraído de bulbos de penas da asa e órgãos de frangos. Essa técnica permitiu a detecção do AGV2 em aves de outros locais na Região Sul do Brasil. O DNA viral foi detectado em 90,7% (98/108) das amostras coletadas no estado do Rio Grande do Sul e 60,4% (29/48) das amostras do estado de Santa Catarina. Os mesmo primers da PCR foram adaptados para examinar tecidos de cérebro de galinhas da Holanda. Nessas amostras o DNA do AGV2 foi detectado em nove das 21 (42,9%) amostras de tecidos cerebrais em aves com lesões hemorrágicas. Essas descobertas fornecem evidências de que infecções de AGV2 são generalizadas e não se restringem a Região Sul do Brasil. Além disso, esses estudos permitiram a identificação de variantes do DNA genômico do AGV2. Análises filogenéticas demostraram que os genomas examinados poderiam ser divididos em três grupos, com base em diferenças nos genes das ORFs das proteínas VP2 e VP3. Em conclusão, a presente tese abrange estudos da identificação de um vírus aviário, até então desconhecido, denominado girovírus aviário tipo 2, que encontra-se amplamente distribuído. A associação deste agente com enfermidades em aves será tema de estudos que estão sendo realizados. / This thesis concerns studies carried out in search for new viral agents in commercial poultry flocks. In the first chapter, the identification of the genome of avian gyrovirus type 2 (AGV2), a new Gyrovirus, is reported. The viral genome the 2383-nucleotide sequence was amplified from sera of commercial broilers by random DNA amplification. Sequence analysis of the amplified product showed that the putative viral sequence had about 40% nucleotide similarity with the genome of its closest relative, chicken anemia virus (CAV). Such low degree of nucleotide similarity justified its classification as a new virus type within the genus, to which the name avian gyrovirus type 2. The amino acid similarity between the predicted viral proteins VP1, VP2 and VP3 of AGV2 and those of CAV was 38.8%, 40.3%, and 32.2%, respectively. In order to examine the amplitude of dissemination of this agent, it became necessary to carry out a search for AGV2 genomes in poultry flocks from other regions. To achieve such objective, an AGV2-specific PCR was designed to amplify viral DNA from nuclei acid extracted from poultry feather shafts. This allowed detection of AGV2 genomes in flocks from other locations in Southern Brazil. Viral DNA was detected in 98/108 (90.7%) of samples collected in the state of Rio Grande do Sul and 29/48 (60.4%) of the samples collected in the state of Santa Catarina. The same PCR primers were adapted to examine brain tissues of chickens from the Netherlands. In such samples, AGV2 DNA was detected in nine out of 21 (42.9%) brain tissue samples from birds with haemorragic lesions. These findings provided evidence that AGV2 infections are widespread and are not restricted to Southern Brazil. In addition, these studies allowed the identification of genomic variants of AGV2. Phylogenetic analyses demonstrated that such genomes could be divided into three clusters. In conclusion, this thesis encompasses studies on the identification of a previously unreported virus, named avian gyrovirus 2, which was later shown to be widely distributed. The relationship between this agent and disease in poultry will be subject of further studies which are being performed in continuation to this work.

A presente tese versa sobre estudos realizados visando a identificação de novos agentes virais em frangos comerciais. No primeiro capítulo, a identificação do girovírus aviário tipo 2 (AGV2) é reportada. Um genoma viral de 2383 nt foi amplificado a partir soros de frangos comerciais por amplificação randômica de DNA. A análise da sequência do produto amplificado, revelou que cerca de 40% das sequencias de nucleotídeos apresentavam similaridade com o genoma do vírus da anemia infecciosa das galinhas (CAV). Dado o baixo grau de identidade com o CAV, o genoma identificado justificou a proposição de um novo tipo de vírus, dentro do gênero gyrovírus foi denominado “girovírus aviário tipo 2” (AGV2). O genoma do AGV2 tem organização semelhante a do CAV, com percentagens de similitude de aminoácidos nas regiões codificantes, correspondentes às proteínas virais VP1, VP2 e VP3 do CAV de 38,8%, 40,3% e 32,2%, respectivamente. A fim de analisar a amplitude da disseminação deste agente, foi realizado um estudo para identificar a ocorrência do AGV2 em granjas de frangos de outras regiões. Para atingir esse objetivo, uma PCR específica ao AGV2 foi desenvolvida para amplificar o DNA viral extraído de bulbos de penas da asa e órgãos de frangos. Essa técnica permitiu a detecção do AGV2 em aves de outros locais na Região Sul do Brasil. O DNA viral foi detectado em 90,7% (98/108) das amostras coletadas no estado do Rio Grande do Sul e 60,4% (29/48) das amostras do estado de Santa Catarina. Os mesmo primers da PCR foram adaptados para examinar tecidos de cérebro de galinhas da Holanda. Nessas amostras o DNA do AGV2 foi detectado em nove das 21 (42,9%) amostras de tecidos cerebrais em aves com lesões hemorrágicas. Essas descobertas fornecem evidências de que infecções de AGV2 são generalizadas e não se restringem a Região Sul do Brasil. Além disso, esses estudos permitiram a identificação de variantes do DNA genômico do AGV2. Análises filogenéticas demostraram que os genomas examinados poderiam ser divididos em três grupos, com base em diferenças nos genes das ORFs das proteínas VP2 e VP3. Em conclusão, a presente tese abrange estudos da identificação de um vírus aviário, até então desconhecido, denominado girovírus aviário tipo 2, que encontra-se amplamente distribuído. A associação deste agente com enfermidades em aves será tema de estudos que estão sendo realizados. / This thesis concerns studies carried out in search for new viral agents in commercial poultry flocks. In the first chapter, the identification of the genome of avian gyrovirus type 2 (AGV2), a new Gyrovirus, is reported. The viral genome the 2383-nucleotide sequence was amplified from sera of commercial broilers by random DNA amplification. Sequence analysis of the amplified product showed that the putative viral sequence had about 40% nucleotide similarity with the genome of its closest relative, chicken anemia virus (CAV). Such low degree of nucleotide similarity justified its classification as a new virus type within the genus, to which the name avian gyrovirus type 2. The amino acid similarity between the predicted viral proteins VP1, VP2 and VP3 of AGV2 and those of CAV was 38.8%, 40.3%, and 32.2%, respectively. In order to examine the amplitude of dissemination of this agent, it became necessary to carry out a search for AGV2 genomes in poultry flocks from other regions. To achieve such objective, an AGV2-specific PCR was designed to amplify viral DNA from nuclei acid extracted from poultry feather shafts. This allowed detection of AGV2 genomes in flocks from other locations in Southern Brazil. Viral DNA was detected in 98/108 (90.7%) of samples collected in the state of Rio Grande do Sul and 29/48 (60.4%) of the samples collected in the state of Santa Catarina. The same PCR primers were adapted to examine brain tissues of chickens from the Netherlands. In such samples, AGV2 DNA was detected in nine out of 21 (42.9%) brain tissue samples from birds with haemorragic lesions. These findings provided evidence that AGV2 infections are widespread and are not restricted to Southern Brazil. In addition, these studies allowed the identification of genomic variants of AGV2. Phylogenetic analyses demonstrated that such genomes could be divided into three clusters. In conclusion, this thesis encompasses studies on the identification of a previously unreported virus, named avian gyrovirus 2, which was later shown to be widely distributed. The relationship between this agent and disease in poultry will be subject of further studies which are being performed in continuation to this work.