Made available in DSpace on 2014-08-20T14:31:28Z (GMT). No. of bitstreams: 1 dissertacao_eliane_machado_pereira.pdf: 4235511 bytes, checksum: 28dcd80ec8888a8603565a7809054107 (MD5) Previous issue date: 2012-03-15 / This study verified helminth parasites infection in gastrointestinal tract, lungs, heart, and kidneys of southern sea lions, Otaria flavescens, from south coast of Rio Grande do Sul State, Brazil. Twenty-nine sea-lions were found dead on the beaches, whose carcasses had mild state of decomposition, were necropsied. The organs were collected during field necropsies for laboratory analyses were 24 small and large intestines, 24 livers including parenchyma and gall bladder, 29 stomachs, 24 hearts, and 24 pairs of kidneys. The organs were maintained frozen at -20°C until their processing. A sieve with 150µm mesh was used for screening the parasites. All content retained was analyzed under stereomicroscope. The helminthes were collected, counted, fixed in AFA, stained with carmine, and clarified in beechwood creosote. The small intestines were divided into three segments that were separately analyzed to record the distribution of helminthes by preference sites. Kolmorogov-Smirnov test was used to verify the type of data distribution. Comparison of mean abundance of infection between age classes was performed through Wilcoxon test at significant level of 0.05. Correlations between infection intensity, sex, total length of the individual, and length of small and large intestines were determined using Pearson s Correlation. Action® software version 1.1 was applied for statistical analyses. Among 29 specimens of O. flavescens 23 were males, three females, and three individuals whose sex could not be determined. The average length of the animals was 2.14±0.31m (1.58 to 2.64m) including 13 sub-adults and 16 adults. A total of 996 specimens of Contracaecum ogmorhini were recorded, especially in the stomach (10.34% of prevalence), 42,145 specimens of Corynosoma australe (100% of prevalence) and 512 of Bolbosoma turbinella (50% of prevalence) were found. Two species of trematodes were found: Stephanophrora uruguayense (Prev. 4.17%) and Ascocotyle (Phagicola) longa (Prev. 33.33% ), a estimated total of 1,988.202 specimens. Cestodes were found in only one of the hosts (4.16% of prevalence) which presented four scoleces. Macroscopically, liver, gall bladder, heart, lungs, and kidneys did not contain parasites. No significant correlation was observed between infection intensity, mean abundance, sex, total length of the host, or length of intestines. Infections levels were similar between sub-adults and adults sea lions. This is the first record of Diphyllobothrium sp., Bolbosoma turbinella, Contracaecum ogmorhini, Ascocotyle (Phagicola) longa, and Stephanoprora uruguayense in O. flavescens in Brazilian waters. As regards parasite fauna of O. flavescens, our data are different from those previously reported for specimens from Pacific coast of South America. / Este estudo analisou os helmintos parasitos gastrointestinais, pulmonares, cardíacos e renais do O. flavescens no litoral sul do Rio Grande do Sul, Brasil. Foram necropsiados 29 leões-marinhos, cujas carcaças apresentavam baixo estado de decomposição. Os órgãos coletados mediante necropsia a campo para análise em laboratório foram 24 intestinos delgado e grosso, 24 fígados incluindo parênquima e vesícula biliar e 29 estômagos, 24 corações e 24 pares de rins. Os órgãos foram congelados a 20oC até o seu processamento. Para a triagem dos parasitos foi usada peneira com malha de 150 µm e todo o conteúdo retido foi analisado sob microscópio estereoscópico. Os helmintos foram fixados em AFA, corados com Carmin e clarificados com creosoto de Faia. Os intestinos delgados foram divididos em três segmentos que foram analisados separadamente para registrar a distribuição dos helmintos por sítios de preferência. Teste de Kolmorogov-Smirnov foi utilizado para verificar tipo de distribuição dos dados. A comparação da abundância média de infecção entre classes etárias foi realizada através do teste de Teste de Wilcoxon usando nível de significância de 0.05. As correlações entre intensidade de infecção, sexo, comprimento total do individuo e comprimento dos intestinos delgado e grosso foram verificadas usando a Correlação de Pearson. Para as análises estatísticas usou-se o software Action® versão 1.1. De 29 espécimes de O. flavescens 23 eram machos, três fêmeas e em três indivíduos o sexo não pode ser determinado. A média do comprimento total dos animais foi 2,14±0,31m (1,58 - 2,64m), sendo 13 subadultos e 16 adultos. Foram registrados 996 espécimes de Contracaecum ogmorhini presentes principalmente no estômago, (prevalência 10%). Registrou-se 42.145 espécimes de Corynosoma australe (Prev. 100%) e 512 de Bolbosoma turbinella (Prev. 50%). Duas espécies de trematódeos foram coletados: Stephanophrora uruguayense (Prev. 4.17% ) e Ascocotyle (Phagicola) longa (Prev. 33.33% ) totalizando 1.988.202 espécimes. Cestódeos foram encontrados em apenas um hospedeiro (Prev. 4,16%) que apresentou 4 escóleces. Macroscopicamente, fígado, vesícula biliar, coração, pulmões e rins examinados não estavam parasitados. Nenhuma correlação significativa foi observada entre a intensidade de infecção, sexo, comprimento total ou comprimento dos intestinos. Este é primeiro registro de Diphyllobothrium sp., Bolbosoma turbinella e Contracaecum ogmorhini em O. flavescens em águas brasileiras. No que se diz respeito à fauna parasitária de O. flavescens, as espécies encontradas no presente estudo não são as mesmas previamente citadas para leãomarinho-do-sul da costa pacífica da América do Sul.

A caracterização de sequências codificadoras presentes nas vizinhanças do gene META 1 permitiu a identificação de alguns genes expressos preferencialmente em estágios infectivos de L. (L.) amazonensis. Um dos genes presentes é regulado de forma distinta, observando-se maior abundância do RNA em formas amastigotas. Este gene foi denominado LaLRR17 por codificar uma proteína contendo em sua região central 6 repetições ricas em leucina (LRR). As LRR são motivos presentes em diversas famílias de proteínas e são responsáveis pela formação de uma estrutura capaz de estabelecer interações protéicas. A região central da proteína LRR17 apresentou similaridade com a porção carboxi-terminal da proteína NOD 3 humana. A proteína LRR17 foi localizada no citosol de macrófagos infectados com L. (L.) amazonensis. Para caracterizar a função da proteína LRR17 foram obtidas linhagens de L. (L.) amazonensis expressoras do gene LmjLRR17. Essas linhagens mutantes foram mais infectivas em macrófagos in vitro quando comparadas com a linhagem selvagem. Avaliamos também o papel das proteínas NOD 1 e NOD 2 na infecção por L. (L.) amazonensis e L. (L.) major para estabelecer a possível relação da proteína LRR17 na interação com essas vias de defesa celular do macrófago. / The characterization of coding sequences in the vicinity of the META 1 gene allowed the identification of some genes preferentially expressed in L. (L.) amazonensis infective stages. One of the identified transcripts presents a distinct pattern of expression with higher levels of mRNA in amastigotes. This gene was named LaLRR17 since it encodes a 72 kDa protein with 6 leucine-rich repeats (LRR) in its central region. Leucine-rich repeats (LRR) are present in several families of proteins and are responsible for the formation of a structure capable of establishing protein interactions. The central region of the LRR17 protein showed similarity with the carboxyl-terminal portion of the NOD 3 human protein. The LaLRR17 protein is secreted to the cytoplasm of L. (L.) amazonensis-infected macrophages. To characterize the function of the LRR17 protein we obtained strains of L. (L.) amazonensis expressing the LmjLRR17 gene. These mutant strains were more infective to macrophages in vitro when compared with the wild type strain. We also evaluated the role of NOD 1 and NOD 2 proteins in infections with L. (L.) amazonensis and L. (L.) major to investigate a possible role of the LRR17 protein in the interaction with these defense pathways in macrophages.

No sistema imune de artrópodes, em contraste com o dos vertebrados, não ocorre uma resposta a antígenos por meio da produção de imunoglobulinas específicas contra agentes infecciosos. Portanto, a imunidade adaptativa está ausente, sendo que o sistema imune nesses animais baseia-se somente numa resposta inata. Os mecanismos presentes na resposta imune inata dos artrópodes incluem: (i) o sistema de coagulação; (ii) a cascata da profenoloxidase; (iii) a liberação de moléculas que possuem ação direta contra microorganismos dentre elas, os peptídeos antimicrobianos (PAMs). Estas moléculas podem ser constitutivas, como observado em aracnídeos, ou terem sua expressão induzida após desafio imune, como observado em alguns insetos. PAMs são moléculas anfipáticas, geralmente catiônicas e compostas por 9 a 100 resíduos. O estudo desses peptídeos, além de possibilitar a descoberta de novas moléculas que tenham modo de ação alternativo aos antibióticos convencionais, permite uma compreensão mais ampla do sistema imunológico de diferentes grupos de animais, bem como a origem dos seus mecanismos na história evolutiva. Neste trabalho foi utilizado o opilião Acutisoma longipes como modelo experimental para a caracterização de PAMs presentes na sua hemolinfa, tendo em vista que não há registros de estudos dessa natureza que utilizaram representantes da ordem Opiliones. Primeiramente, foi demonstrada a ocorrência de diversas frações, obtidas da purificação da hemolinfa, com atividade anti-M. luteus, consistidas por peptídeos que aparentemente são constitutivos. Uma das frações mostrou-se pura e apresentou um peptídeo de 2,1 kDa cuja estrutura primária, composta por 18 resíduos, foi completamente elucidada (SGYLPGKEYVYKYKGKVF) por sequenciamento de novo e sequenciamento do N-terminal. Este peptídeo linear foi nomeado longipina. O peptídeo sintético apresentou atividade antimicrobiana contra as bactérias Escherichia coli e Micrococcus luteus, e as leveduras Candida albicans e C. tropicalis, além de não apresentar atividade hemolítica na máxima concentração testada (100 mM). Foi demonstrado que a longipina liga-se preferencialmente a vesículas unilamelares grandes (LUVs), constituídas de fosfolipídios aniônicos (POPG), que mimetizam as cargas da superfície das células de microorganismos. A ligação do peptídeo a LUVs compostas, na razão molar de 1:1, por POPG e POPC (zwiteriônico) provoca o extravasamento de marcadores (carboxifluoresceína) aprisionados nos seus interiores. Esse peptídeo encontra-se majoritariamente desestruturado em solução ou na presença de sistema mimético composto somente por POPC. Porém, na sua ligação com POPG:POPC 1:1, adota estruturas b e estrutura relacionada à presente em agregados intermoleculares de fibras amiloides. / The arthropods immune system, in contrast to the vertebrates one, lacks a response to antigens through the production of specific immunoglobulins to fight against infectious agents. Therefore, the adaptive immunity is absent, and their immune system is only supported by an innate response. The mechanisms present in the arthropods innate immune response include: (i) the clotting system; (ii) the prophenoloxidase cascade; (ii) the release of molecules that can directly act against microorganisms among them, the antimicrobial peptides (AMPs). These molecules can be constitutive, as observed in arachnids, or have their expression induced upon imune challenge, as observed in some insects. AMPs are amphipathic molecules, usually cationic and formed by 9 to 100 residues. The study of these peptides, besides allowing the discovery of new molecules that exert their mode of action alternatively from the conventional antibiotics, permits a wider understanding of the immunological system from different groups of animals, as well as the origin of their mechanisms in the evolutionary history. The harvestman Acutisoma longipes was used in this work as na experimental model for the characterization of AMPs from its haemolymph, taking into account that there are no records of using representatives of the Opiliones order in such kind of study. Firstly, it was demonstrated the occurrence of several fractions, obtained from the haemolymph purification, that presented anti-M. luteus activity, consisted by peptides that are apparently constitutive. One of these fractions was pure and presented a 2.1 kDa peptide which had its primary structure, composed by 18 residues, completely elucidated by de novo and N-terminal sequencing (SGYLPGKEYVYKYKGKVF). This linear peptide was named longipin. The synthetic peptide presented antimicrobial activity against Escherichia coli and Micrococcus luteus bacteria and Candida albicans and C. tropicalis yeasts, in addition to the absence of hemolytic activity at the highest concentration used (100mM). It was shown that longipin preferentially binds to large unilamellar vesicles (LUVs), constituted by anionic phospholipds (POPG), that mimics the membrane surface of microbe cells. The peptide biding to LUVs composed, in 1:1 molar ratio, by POPG and POPC (zwitterionic) causes dye leakage (carboxyfluorescein) from their inside. This peptide in solution or in the presence of POPC solely containing mimic system is mostly unstructured. However, upon its binding to POPG:POPC 1:1, it adopts b and intermolecular aggregates amyloid-like fibrils structures.

Malaria continues to be a challenging problem in the developing world, and the burden of this life threatening disease continues to be borne by young children living in Sub Saharan Africa. One of the biggest challenges to the prevention and control of this problem lies in accurately diagnosing malaria, and distinguishing it from the many other febrile illnesses which present in children in this age group. Liberia is a West African country with a high burden of malaria. Very little is known about the presentation of severe malaria in children aged 5 years old and younger in Liberia. We undertook a prospective, hospital -based study of children 5 and under presenting to JKF Medical Center, the national referral hospital, with fever and signs and symptoms consistent with malaria. The aims of our study were to determine: 1) the frequency of confirmed malaria cases, 2) the frequency of non-malaria diagnoses, 3) the prevalence of anti-malarial drug resistance mutations, 4) the presence of other life threatening etiologies of febrile illness such as S. typhii and Dengue virus and 5) immunological profiling associated with severe malaria. We analyzed clinical and laboratory data from 462 children age 5 and under who presented to the national referral hospital in Monrovia, Liberia with signs and symptoms consistent with malaria over a one year period. Key findings included determining the demographic factors most closely associated with severe malaria in this population (age > 1yr and urban environment) and those that were negatively associated with the development of severe malaria (prior episodes of malaria, use of bednets and use of anti malarial medications prior to presentation). The clinical symptoms most closely associated with severe malaria in this population were found to be headache and vomiting. We found that 33% of children admitted and treated for severe malaria did not test positive for malaria by rapid diagnostic testing (RDT) or blood smear. These children had a case fatality rate that was 5 times higher than their RDT positive counter parts. Of the RDT negative children, 2 tested positive for salmonella typhii, but were not treated for this pathogen. Upon discharge from the hospital, 11% of children had resolved their symptoms, but had not cleared their malaria parasites. These findings will help to identify the children who present with true severe malaria in Liberia. They also underscore the need to expand diagnostic capabilities to determine which other types of pathogens cause febrile illness in this population, so that adequate treatment can be extended to these patients. The immunoprofiles of these children revealed 3 IgM antibodies (AMA-1, CSP and LSA-1) that were associated with the development of severe malaria. These antibodies also appear to be associated with initial infection with malaria. Such data will help to identify antigens could be potential targets for malaria vaccines, and which can play an important role in the development of new malaria diagnostics for this population.

This thesis deals with computing of the magnetic lens with a perturbed pole piece due to manufacturing imperfections. Two possible ways of calculation are discussed - the perturbation theory and 3D computing. Three methods for evaluating axial multipole field functions from 3D fields are introduced. Beam spots in the image plane and aberration coefficients are computed and results obtained by the application of perturbation theory are compared to results evaluated from 3D simulations. Consequently, a suitability of using the perturbation theory is discussed.

Fast switching operations in IGBTs generate electromagnetic field disturbances, which might cause EMI and functionality issues. For higher frequency characterization, the parasitic inductances and capacitances have to be considered. The characterization of the electromagnetic field disturbances in- and around the converter module could be predicted early in the design. The study involves a high frequency characterization of electric fields (Efield), magnetic fields (H-fields) and the surface currents distribution in a converter module. The high frequency electromagnetic software (CST) was used for the analysis. A given 3D CAD model of an AC/DC converter module was analyzed in CST. The CAD contained IGBT bus-bars interconnections, converter casing, heat sink and other metallic structures. The ACside has six IGBTs and the DC-side has a chopper which has two switches. The IGBTs ONstate and OFF state was modeled with lumped elements. The DC link capacitor was just modeled as lumped elements, while the metallic capacitor casing was included in the 3D model for analyzing the field distribution inside the converter casing. To check the model accuracy, CST models were compared with PEEC (Partial Element Equivalent Circuit) models for simple antenna cases. Using the converter geometry, CST estimates the parasitics and the eventual current, voltage and electromagnetic field distributions for a given excitation signal. The DC-link was excited with a step pulse and the fields were computed. With consideration of specific design details, the modeling approach developed in this study, could be used to construct high frequency models of converter modules for different projects.

In recent years, multi-functional reconfigurable antennas (MRA) has attracted much attention in wireless communication. The reconfigurable antenna can adapt itself with changing system conditions, and it can provide different multi-functionalities which can give better system performances. Instead of using multiple antennas, a single reconfigurable antenna can provide the same performance and occupy less space. By using the parasitic layer technique, an antenna can be turned into a reconfigurable antenna. The main objective of this thesis is to study radiation pattern reconfiguration of the horn antenna by using the parasitic layer concept. The MRA consists of a single horn, dielectric loaded truncated pyramid (DLTP), and the parasitic layer. The antenna that is chosen in this thesis is the horn antenna because it provides high directivity. DLTP is used for magnification purpose. The results show that three modes of operations that provide better performances compared to the single horn antenna are achieved.

Anion intercalation in the graphite positive electrode of a dual-ion battery requires high potential (> 4.3 V vs Li+/Li), which aggravates parasitic reactions involving electrolyte decomposition and Al corrosion, manifesting in poor coulombic efficiency, cycle life, and quick self-discharge. This study aims to investigate the stability of anion-intercalated graphite electrodes in a 4 M solution of lithium bis(fluorosulfonyl)imide (LiFSI) in ethyl methyl carbonate (EMC) using both in-situ and ex-situ Raman spectroscopy. The concentrated electrolyte is essential as it limits parasitic reactions at the cathode-electrolyte interface (CEI) occurring in parallel to anion intercalation. Using electrochemical methods including cyclic voltammetry, and post-mortem electron microscopy it was confirmed that the Al current collector is largely stable at potentials as high as 5.2 V in the electrolyte under consideration; no dissolved Al species were detected using EDX characterization. Results from the cyclic voltammetry study also indicate that parasitic reactions can be mitigated when the cut-off potential is limited to 5.0 V leading to higher coulombic efficiency (CE = 94 %) and more stable discharge capacity (85.17 mAh g-1). However, extending the potential to 5.1 and 5.2 V results in the discharge capacity increasing by almost 20 mAh g-1, though at the expense of the coulombic efficiency, which decreases from 94 to 76 %. Upon raising the cut-off potential to 5.3 V, the CE significantly decreased (20.62 %) as a result of extensive solvent decomposition ultimately leading to much quicker capacity fading.  Based on SEM images taken after 50 cycles, graphite particles did not sustain any structural or morphological change during cycling regardless of the cut-off potentials applied. Further tests were conducted on Li-graphite DIBs using galvanostatic methods in the range from 3 to 5 V, and at different specific currents (20, 50, and 100 mA g-1). Though the cells exhibited good performance in terms of capacity retention, and cycle life at all currents, the coulombic efficiency tended to decrease as the test currents were lowered. This observation confirms the presence of parasitic reactions which are only visible when the experimental timescale is sufficiently long. At 50 and 100 mA g-1, the CE reached > 98 % which further verifies the kinetic aspect of electrolyte decomposition reactions. It is evident that self-discharge sustained in the course of open-circuit potential (OCP) relaxation of the fully charged cell can reveal the stability of the electrolyte and the anion-intercalated graphite. Raman spectroscopy measurements conducted in-situ and ex-situ on graphite electrodes charged and discharged to a series of potential cut-offs reveal the existence of self-discharge leading to extraction of anions from the graphite particles. This was demonstrated through the spectral appearance of E2g2(i) band next to E2g2(b) band at a fully intercalated state, as opposed to the in-situ spectrum, which only showed one intercalated band (E2g2(b)). It can be concluded that concentrated electrolytes (such as 4 M LiFSI in EMC) only provide kinetic stability and are unable to entirely inhibit parasitic reactions at the interface. This further highlights the need for electrolyte additives that can create a more stable interfacial passivation layer on the positive electrode so that more reversible anion intercalation can be attained.

The majority of organic waste globally is either dumped or placed on landfills, which can result in spreading of disease and pests, methane gas emissions, the deterioration of landscapes and odour pollution. One of the solutions to poor organic waste management is to create value from waste. A way to do this is with black soldier fly (Hermetia illucens) larvae (BSFL) composting. The interest for BSFL composting has increased the past 10 years mainly because it is a technology with relatively low investment costs. The part of the process where the seed larvae that are used in the treatment are produced is called BSF-colony. Producing seed larvae in an efficient way is an important part of the technology’s feasibility. However, there is limited research published on BSF-colony management, especially on a larger scale. The aim of the study was to investigate the factors that impact the efficiency of the BSF-colony on a large scale BSFL composting company in Kenya in a semi-open setting. The efficiency of the BSF-colony management was assessed in terms of emergence rate (percentage of pupa that emerge as flies), hatching rate (percentage of eggs that hatch and survive to 5 day old larva) and number of eggs laid per female BSF. The result of the study suggests that parasitic wasps (Dirhinus giffardii) can reduce the emergence rate significantly, and high temperatures and water shortage for the adult BSF can reduce the egg production. Observations made during the study indicate that personnel routines and how the BSF-colony is arranged also could affect the egg production significantly. The overall variation in the results suggests that other factors, beyond the ones investigated in the study, impact the efficiency of the BSF-colony, and further research regarding BSF-colony management is recommended / Merparten av det organiska avfallet i världen dumpas eller läggs på deponier, vilket kan leda till spridning av sjukdomar och skadedjur, utsläpp av metangas, förstörelse av landskap och luktföroreningar. En av lösningarna på bristfällig hantering av organiskt avfall är att skapa värde av avfallet. Ett sätt att göra det är med hjälp av fluglarvskompostering med amerikansk soldatfluga (Hermetia illucens). Intresset för fluglarvskompostering har ökat under de senaste tio åren, främst på grund av att det är en teknik som inte behöver innebära höga investeringskostnader. Flugkolonin är den del av processen där sättlarverna som användes i komposteringen produceras. Att sättlarver produceras på ett effektivt sätt är en viktig del av teknikens genomförbarhet. Det finns dock begränsad forskning publicerad om effektivisering av flugkolonier, speciellt sådan som berör storskaliga kolonier. Syftet med studien var att undersöka vilka faktorer som påverkar flugkolonins effektivitet på ett storskaligt fluglarvskomposteringsföretag i Kenya i en halvöppen miljö. Flugkolonins effektivitet bedömdes i termer av metamorfosfrekvens (den andel av pupporna som blir flugor), antalet lagda ägg per fluga och kläckningsfrekvens (den andel ägg som kläcks och överlever till 5-7 dagar gamla larver). Resultatet av studien tyder på att parasitsteklar kan minska metamorfosfrekvensen avsevärt, och att höga temperaturer samt vattenbrist kan minska äggproduktionen. De observationer som gjordes under studien tyder på att personalens rutiner och hur flugkolonin är anordnad också kan påverka äggproduktionen avsevärt. Den relativt stora variationen i alla resultat tyder på att andra faktorer än de som undersökts i studien påverkar flugkolonins effektivitet, fortsatta studier inom effektivisering av flugkolonier rekommenderas

<p>  </p> <p>Several plant-parasitic nematode (PPN) species cause decline in the health of creeping bentgrass putting greens. PPNs target and parasitize the root systems of turfgrass which may exacerbate the impact of other biotic and abiotic stress. Turfgrass managers often apply nematicides preventatively or curatively to control PPN populations. However, the inherent chemistries of the nematicides may inhibit their ability to permeate through the thatch layer and soil, resulting in an ineffective application. This research aimed to evaluate the depth of PPN populations through the growing season to maximize the effectiveness of nematicide applications, with a primary focus on lance (<em>Hoplolaimus</em> spp.) and root-knot nematodes (<em>Meloidogyne</em> spp.). To determine the depth of genera across time, soil samples were taken with a 1.9 cm diameter soil probe to a depth of 25 cm during the months of April, June, August, and October at 7 sites across Missouri, three in eastern Kansas and ten sites in Indiana in 2021 and 2022, respectively. Significant interactions occurred between sampling depth and month in both Missouri and Indiana for some PPN genera. Additionally, individual lance and root-knot nematode species obtained from sampling were characterized with molecular methods and in the case with one lance nematode from Indiana, with scanning-electron microscopy. Results suggest an over-representation of <em>H. galeatus</em> in diagnostic literature, and a diverse collection of <em>Meloidogyne</em> spp. present in Indiana on golf course putting greens. </p>