A variação na disponibilidade de nitrogênio e magnésio pode alterar as características produtivas, nutricionais e metabólicas das plantas forrageiras. O objetivo foi de avaliar o efeito das combinações de doses de nitrogênio e de magnésio na produção da parte aérea e das raízes, nas concentrações dos nutrientes nas lâminas de folhas recém-expandidas (LR) e nas raízes, na atividade da glutamina sintetase, nas concentrações das formas inorgânicas de nitrogênio e na relação nitrogênio:magnésio nas LR do capim-marandu. O experimento foi realizado em casa de vegetação, em Piracicaba SP, com a Brachiaria brizantha cv. Marandu. Foram estudadas cinco doses de nitrogênio (2; 9; 16; 23 e 30 mmol L-1) e cinco doses de magnésio (0,05; 0,70; 1,35; 2,00 e 2,65 mmol L-1) em solução nutritiva, em esquema fatorial 52 fracionado, perfazendo treze combinações: 2/0,05; 2/1,35; 2/2,65; 9/0,70; 9/2,00; 16/0,05; 16/1,35; 16/2,65; 23/0,70; 23/2,00; 30/0,05; 30/1,35 e 30/2,65. O delineamento experimental foi o de blocos ao acaso, com quatro repetições. As plantas tiveram dois períodos de crescimento, nos quais foram avaliadas as produções de massa seca, contados perfilhos e folhas, coletado material fresco para determinação da atividade da glutamina sintetase, determinada a área foliar e determinada as concentrações dos nutrientes e nitrato e amônio nas lâminas das folhas recém-expandidas. Após o segundo corte as raízes foram separadas da parte aérea sendo determinada a massa seca das raízes, o comprimento e superfície radiculares e as concentrações dos nutrientes nas raízes. As combinações de altas doses de nitrogênio e magnésio resultaram em alta produção de massa seca, número de folhas e perfilhos, área foliar, massa seca das raízes, e comprimento e superfície radicular do capim-marandu. As concentrações de nitrogênio (segundo corte) e cálcio (nos dois cortes) nas LR e a concentração de nitrogênio nas raízes aumentaram com as combinações de elevadas doses de nitrogênio e de magnésio. A concentração de potássio nas LR, nos dois cortes, e nas raízes da gramínea foi mais baixa quando se utilizaram combinações de elevadas doses de nitrogênio e magnésio. De forma isolada, o incremento nas doses de nitrogênio resultou em reduzidos comprimento e superfície específicos das raízes, mais alta concentração de nitrogênio nas LR e mais alta concentração de cálcio nas raízes e o incremento das doses de magnésio proporcionou pequenos comprimento e superfície específicos das raízes, mais alta concentração de magnésio nas LR e nas raízes e mais baixa concentração de cálcio nas raízes do capim. A concentração de nitrato nas folhas diagnósticas foi influenciada pelas combinações de doses de nitrogênio e magnésio enquanto a concentração de amônio aumentou com o incremento das doses de nitrogênio. A atividade da glutamina sintetase foi mais baixa em altas doses de nitrogênio, nos dois crescimentos da gramínea. A relação de 6,3:1 entre as concentrações de nitrogênio e de magnésio nas LR correspondeu à máxima produção de massa seca no primeiro corte do capim. / Productive, nutritional and metabolic characteristics of the forage plants may be changed by the nitrogen and magnesium availability. The objective was to study the effects of nitrogen and magnesium rates on marandu palisadegrass growth, by evaluating the shoot and root yield, leaf and root concentrations of nutrients and leaf concentration of inorganic forms of nitrogen, glutamine synthetase activity and nitrogen:magnesium ratio in recently expanded leaf laminae (RL). The experiment was carried out in a greenhouse at Piracicaba, State of São Paulo, Brazil, with the Brachiaria brizantha cv. Marandu. Five nitrogen rates (2, 9, 16, 23 and 30 mmol L-1) and five magnesium rates (0.05, 0.70, 1.35, 2.00 and 2.65 mmol L-1) were used in nutrient solutions, in a fractorial 52 factorial, resulting in the following combinations: 2/0.05, 2/1. 35, 2/2.65, 9/0.70, 9/2.00, 16/0.05, 16/1.35, 16/2.65, 23/0.70, 23/2.00, 30/0.05, 30/1.35 and 30/2.65. A randomized block design, with four replications was used. Plants had a growth and regrowth period. After the second harvest it was evaluated the dry mass yield, number of tillers and leaves, sampled fresh material to assay the glutamine synthetase activity, measured the leaf area, and determined the concentrations of nutrients, nitrate and ammonium in the recently expanded leaf laminae (diagnostic leaves). After the second harvest the roots were separated from the tops and was determined the root dry mass, total length and surface, specific length and surface, and concentrations of nutrients in such plant tissue. High nitrogen and magnesium rates resulted in high dry mass yield, leaf area, number of leaves and tillers, in both growth periods of the grass, as well as in roots dry mass, length and surface. Nitrogen (at the second harvest) and calcium (both harvests) concentrations in diagnostic leaves and nitrogen concentration in the roots was high when high rates of nitrogen nitrogen were combined with high rates of magnesium. Diagnostic leaves, in the two grown periods, and roots sampled had low potassium concentrations in plants growth with high rates of both nitrogen and magnesium. Increases in nitrogen rates increased nitrogen concentration in the diagnostic leaves and calcium concentration in the roots, and decreased root specific length and surface. Increases in magnesium rates increased magnesium concentration in the diagnostic leaves and roots, but decreased root calcium concentration and root specific length and surface. Ammonium concentration was increased by nitrogen rates and nitrate concentration was higher under magnesium deficiency. Glutamine synthetase, determined at the two growth periods, decreased with increasing nitrogen rates. Maximum dry mass yield in the first harvest of the grass was found when the nitrogen and magnesium concentrations ratio was 6.3:1 in the diagnostic leaves.

O Brasil é um dos principais produtores mundiais de Eucalyptus spp., mas a produção dessa cultura tem sido comprometida por perdas causadas pelo fungo Puccinia psidii Winter, agente causal da ferrugem do eucalipto. Compreender os mecanismos moleculares da patogenicidade desse fungo, conhecer a composição química de variedades de Eucalyptus spp. resistentes e suscetíveis ao fitopatógeno e ter em mãos uma ferramenta de diagnóstico precoce da doença são conhecimentos de fundamental importância para o desenvolvimento de estratégias de controle do fitopatógeno. Uma das principais barreiras que limitam o estudo molecular de P. psidii é o fato dessa espécie ser biotrófica obrigatória, tendo seu desenvolvimento in vitro limitado. Estudos de investigação a respeito de fungos biotróficos obrigatórios são realizados principalmente in planta e em estágios tardios da doença, deixando grande parte do conhecimento dos processos iniciais de infecção desconhecidos. Informações a respeito dos estágios iniciais da infecção, nos quais diversas estruturas típicas são formadas e podem indicar importantes características sobre fungos biotróficos, são de difícil acesso em estudos in planta em função da grande quantidade de material vegetal em relação à quantidade de material do fitopatógeno, que não é passível de ser removido completamente das amostras analisadas. Dessa forma, o presente projeto de pesquisa foi realizado com os objetivos de desenvolver um protocolo para induzir a germinação e a morfogênese estrutural in vitro de P. psidii, identificar genes candidatos relacionados à diferenciação das estruturas de infecção e de fatores de virulência, caracterizar os metabólitos presentes nas ceras cuticulares de folhas de Eucalyptus grandis resistentes e suscetíveis à ferrugem do eucalipto e desenvolver uma metodologia sensível e eficaz para detectar, quantificar e monitorar a presença de P. psidii em folhas de Eucalyptus grandis assintomáticas. Os dados obtidos no presente trabalho podem auxiliar a compreensão da interação planta-patógeno durante os estágios iniciais de infecção da ferrugem e colaboram para o entendimento da biologia do fitopatógeno para que no futuro sejam desenvolvidas melhores estratégias de controle de P. psidii em plantios de eucalipto. / Brazilian production of Eucalyptus spp. is one of the greatest in the world but it has been affected by Puccinia psidii Winter, the causal agent of eucalyptus rust. The comprehension of molecular mechanisms of pathogenicity and chemical composition of resistant and susceptible eucalyptus plants as well as having a molecular tool for early detection of the disease in field can be used for the development of improved control strategies against this phytopathogen. Molecular studies of P. psidii is limited because it is an obligate biotrophic fungus with limited in vitro development. Biotrophic fungi investigations are made mainly in planta at late developmental stages of the disease. This way, most information of early stages of infection as the development of specialized structures of biotrophic fungi are little understood. The study of early stages of the infection process of biotrophic fungi in planta is hampered by the high amount of plant material in relation to fungi material which is difficult to be obtained in an isolated form for analysis. In this work we developed a protocol to increase in vitro germination and structural differentiation of P. psidii and used this protocol to obtain isolated fungi material for the identification of candidate genes related with virulence factors and initial structural morphogenesis. Moreover, we analyzed the composition of metabolites present in the cuticular wax of leaves from resistant and susceptible E. grandis plants and developed a methodology for the detection of P. psidii in asymptomatic leaves of Eucalyptus grandis. Data obtained in this work help the comprehension of E. grandis-P.psidii interaction at early stages of the infection process and can be used for the development of improved control strategies of eucalyptus rust.

Dentre os diversos fatores que afetam a produção e qualidade de frutos em plantas cítricas, os fatores fitossanitários destacam-se pela grande importância. Entre as diversas doenças que afetam a cultura dos citros, a Clorose Variegada dos Citros (CVC) é uma das mais severas e destrutivas, provocando tanto perdas em produção quanto em qualidade. Pelo fato desta doença ser causada por uma bactéria que se aloja no xilema, toda a dinâmica da água e nutrientes é afetada, resultando em uma série de sintomas visuais em diferentes graus. Buscou-se avaliar o estado nutricional de plantas cítricas sem sintomas vis uais de CVC e, com sintomas em diferentes níveis, através do Critério de Faixas de Suficiência, tradicionalmente utilizado na Citricultura Paulista e, através do Sistema Integrado de Diagnose e Recomendação, denominado pela sigla DRIS. O estudo foi realizado em pomares comerciais de laranjeira 'Pera' enxertada em limão 'Cravo', com sete anos de idade, no município de Araraquara - SP. Foram selecionadas em dois talhões, plantas com três níveis de CVC e plantas sem sintomas visuais, determinando-se os teores dos macronutrientes (N, P, K, Ca, Mg, S) e dos micronutrientes (B, Cu, Fe, Mn e Zn) em folhas de ramos frutíferos, bem como produção de frutos de cada planta. A interpretação através do método DRIS, envolveu a utilização de um "software" aplicado especialmente para a cultura dos citros pelo Instituto Agronômico de Campinas - IAC, onde foi utilizado o método de cálculo proposto por Jones (1981). O DRIS foi eficiente, possibilitando a ordenação dos nutrientes, e, indicando deficiência nutricional para alguns nutrientes considerados adequados ou excessivos pelo Critério de Faixas de Suficiência. Independentemente da presença de sintomas de CVC, todas as plantas apresentaram baixos teores de Ca, resultando em diagnose de deficiência pelos dois critérios de interpretação; de modo geral, houve uma redução nos teores foliares de N e P, em plantas com sintomas mais severos. Com relação aos teores dos demais nutrientes não houve um padrão de decréscimo ou aumento em função da CVC. O "software" DRIS aplicado especialmente para a cultura dos citros pelo IAC, com a população de referência por ele utilizada, deve ainda sofrer ajustes, de acordo com as condições e objetivos do trabalho para melhor correlação com produtividade. / Among all factors which affect the yield and fruit quality from citric plants, pests and diseases are of outstanding importance. There are several pests and diseases wich affect the citrus crop. The citrus variegated chlorosis (CVC) is one of the most severe and destructive diseases affecting both yield and quality. Because this disease is caused by a xylem bacteria, the water and nutrients dinamics is affected resulting in a series of visual simptons of different intensities. There objective of this work was to evaluate the nutritional status os citrus plants without visual simptons of CVC and, with different levels of simptons of this disease. The Sufficiency Range Criteria, traditionally used in the state of São Paulo and the Diagnosis and Recommendation Integrated System (DRIS) were the methods used for leaf analysis interpretation. The study was carried out in commercial orchards of 'Pêra' sweet orange budded on 'Cravo' lemom, seven years old, in the region of Araraquara - SP. Plants with three levels of CVC and plants without visual simptons were selected from two plots. Concentrations of macronutrients (N, P, K, Ca, Mg, S) and micronutrients (B, Cu, Fe, Mn, Zn) were determined on leaves of fruit branches. Fruit production from each plant was measured. The analysis through the DRIS method involved the utilization of a software developed specially for the citrus crop by the Instituto Agronômico de Campinas - IAC, where the method proposed by Jones (1981) was used. The results showed that the DRIS was efficient in ranking the nutrients and indicating the nutritional defficiency for some nutrients considered adequate or excessive by the Sufficiency Range Criteria. Regardless the presence of CVC simptons, all plants show low foliar concentrations of Ca, wich resulted in a diagnose of defficiency by the two criteria of interpretation. There was a foliar concentration reduction of N and P, in plants with more severe simptoms. With respect to the foliar concentration of the other nutrients there was not observed a standard decrease or increase due to CVC. The local DRIS-Citros IAC software norms utilized should be adjusted according to the work conditions and objectives to a improve yield correlation with plant nutrition.

Orientador: Arthur Bernardes Cecílio Filho / Resumo: Em solos onde o cultivo de hortaliças e a aplicação de fertilizantes minerais são constantes, os teores dos nutrientes nestes locais podem aumentar consideravelmente, principalmente fósforo (P) e potássio (K), que estão entre os três nutrientes mais aplicados nas culturas agrícolas. Assim, objetivou-se avaliar a resposta da beterraba a doses de fósforo e de potássio em Latossolo com altos teores destes nutrientes. Foram realizados dois experimentos em campo, o delineamento experimental de ambos foi em blocos ao acaso, em esquema fatorial 5 x 2 e 4 x 2 para os experimentos de P e de K, respectivamente, com quatro repetições. Em ambos experimentos foram utilizadas duas cultivares ('Early Wonder' e 'Kestrel'). As doses de P foram 0, 120, 240, 360 e 480 kg ha-1 de P2O5; e as doses de K foram 0, 60, 120 e 180 kg ha-1 de K2O. Aos 50 dias após o transplante avaliaram-se os teores de nitrogênio, fósforo, potássio e boro na folha diagnose. Produtividade e acúmulo de nutrientes foram feitos na colheita. Nos dois experimentos, teores de P e K no solo eram altos. Os acúmulos para todos os nutrientes avaliados na ‘Early Wonder’ na parte aérea e na raiz tuberosa foram superiores a 'Kestrel' em ambos experimentos. Não foi observado efeito das doses de P e de K na produtividade. Em Latossolo com teor alto de P e de K disponível, as cultivares ‘Early Wonder’ e 'Kestrel' não respondem de forma diferenciada à adubação fosfatada e potássica com relação à produtividade, portanto, não se recomenda... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In soils where vegetable cultivation and mineral fertilizer application are constant, nutrient levels at these sites can increase considerably, especially phosphorus (P) and potassium (K), which are among the three most commonly applied nutrients in agricultural crops. Thus, the objective was to evaluate the response of beet to phosphorus and potassium doses in Oxisol with high levels of these nutrients. Two field experiments were carried out and the experimental design was randomized blocks in a 5 x 2 and 4 x 2 factorial scheme for the P and K experiments, respectively, with four replications. In both experiments two cultivars ('Early Wonder' and 'Kestrel') were used. The doses of P were 0, 120, 240, 360 and 480 kg ha-1 of P2O5; and the K rates were 0, 60, 120 and 180 kg ha-1 of K2O. At 50 days after transplantation, the levels of nitrogen, phosphorus, potassium and boron in the diagnosis leaf were evaluated. Productivity and nutrient accumulation were measured at harvest. In both experiments, soil P and K contents were high. The accumulations for all nutrients evaluated in 'Early Wonder' in the shoot and root were higher than 'Kestrel' in both experiments. No effect of P and K doses on yield was observed. In Oxisol with high content of P and K, 'Early Wonder' and 'Kestrel' cultivars do not respond differently to phosphate and potassium fertilization regarding to yield, so it is not recommended to fertilize beet crop with these nutrients in Oxisol with high levels of both nu... (Complete abstract click electronic access below) / Mestre

In den vorliegenden Studien wurden verschiedene diagnostische Verfahren zum Nachweis von Protozoen und Nematoden im Hinblick auf Sensitivität, Arbeitsaufwand und Kosten miteinander verglichen. Zudem wurde die Eignung der PCR zur molekularen Charakterisierung der Cryptosporidium spp. exemplarisch an Igelkotproben getestet. Bei der Untersuchung von 90 Ferkelkotproben auf I. suis war die Sensitivität eines Kotausstriches mit nachfolgender Autofluoreszenzmikroskopie (AM) signifikant höher als bei einem Flotationsverfahren (FV) mit NaCl-Zucker-Lösung und bei dem kombinierten Sedimentations-Flotations-Verfahren (KSFV) mit verschiedenen Flotationslösungen (NaCl, ZnSO4, NaCl-Zucker-Lösung) mit nachfolgender Lichtmikroskopie. Zudem ist der Arbeitsaufwand für die AM deutlich geringer als bei dem FV und KSFV. Die höheren apparativen Kosten für die AM sind bei hohem Probendurchsatz durch den geringeren Zeitaufwand und der höheren Sensitivität gerechtfertigt. Die Anzahl Kryptosporidien-positiver Proben war bei der Untersuchung von 103 Kälberkotproben auf Cryptosporidium sp. mittels Enzymimmunoassays (EIA; ProSpecT® Cryptosporidium Microplate Assay) im Vergleich zur Karbolfuchsin-Färbung (CF) nach HEINE (1981) und der modifizierten-Ziehl-Neelsen-Färbung (MZN) nach HENRIKSEN u. POHLENZ (1982) am höchsten und signifikant höher als bei der Anwendung der MZN, wenn 10 Blickfelder durchmustert wurden. Bei der Untersuchung von 74 Igelkotproben auf Cryptosporidium sp. mittels EIA (ProSpecT®), einem immunochromatographischen Verfahren (FASTest® CRYPTO Strip), der MZN nach HENRIKSEN u. POHLENZ (1981) und einem direkten Immunfluoreszenz-Test (IFA; MERIFLUOR Cryptosporidium/Giardia) wurden in 9 (EIA), 10 (FASTest®), 11 (MZN) und 12 (IFA) Proben Cryptosporidium sp. nachgewiesen. Der Arbeitsaufwand des FASTest® und der CF ist mit dem EIA vergleichbar, während der IFA und die MZN mehr Zeit benötigen. Die Anwendung des FASTest®, des IFA und des EIA ist mit höheren Kosten verbunden als bei den Färbemethoden, können aber gut in den Arbeitsablauf eines diagnostischen Labors eingefügt werden und sind einfach auszuwerten. Darüber hinaus wurden 45 Kotproben, welche bis zu 27 Tage bei verschiedenen Temperaturen (+6 °C, +16 °C, +30 °C, +40 °C) gelagert wurden, untersucht, um einen Einfluss der Temperatur auf das Untersuchungsergebnis von EIA, CF und MZN zu ermitteln. Während sich die Anzahl positiver Proben bei der Untersuchung mit den Färbemethoden temperatur- und zeitabhängig reduzierte, wurde das Untersuchungsergebnis mittels EIA von der Lagerungstemperatur nicht beeinflusst, so dass ungekühlt transportierte Proben vorzugsweise mit dem EIA untersucht werden sollten. Dagegen ist die CF aufgrund ihrer einfachen und preiswerten Durchführung zur Untersuchung einer hohen Anzahl an Proben geeignet, sofern eine ununterbrochene Kühlung der Proben gewährleistet ist und diese innerhalb von drei Tagen untersucht werden. Der FASTest® ist zur Anwendung in Tierarztpraxen und Ställen geeignet, da zur Untersuchung kein Mikroskop benötigt wird und die Resultate schnell vorliegen. Die Verwendung des IFA, der Kryptosporidien-Oozysten und Giardien-Zysten nachweist, bietet sich vor allem bei Proben an, die auf beide Protozoen untersucht werden sollen. Das Vorkommen der Kryptosporidiose bei unterentwickelten und geschwächten Igeln, welche zum Überwintern in Igelstationen aufgenommen werden, ist hoch. Von 188 untersuchten Igelkotproben konnten in 29,8 % der Proben Cryptosporidium spp. nachgewiesen werden. Durch die Genotypisierung der Kryptosporidien aus 15 positiven Igelkotproben mittels RFLP-PCR basierend auf dem 18S rRNA-Gen konnte in allen untersuchten Proben die Präsenz von C. parvum gezeigt werden. Mit Hilfe der Multilocus-Sequenz-Typisierung der Fragmente des 60kDa Glycoprotein-Gens, des 18S rRNA-Gens, des Actin-Gens und des 70 kDa Hitzeschockprotein-Gens konnten drei verschiedene Subtypen-Familien (IIa, IIc und eine neue als VIIa vorgeschlagene Subtypen-Familie) erkannt werden. Die von den Igeln ausgeschiedenen Kryptosporidien-Oozysten mit zum Teil nachgewiesenem zoonotischen Potential (IIa Subtypen-Familie) könnten eine Infektionsquelle für den Menschen sein, aber auch ein antropozoonotisches Potential (IIc Subtypen-Familie) sollte in Betracht gezogen werden, so dass die Hygiene in den Igelstationen einen hohen Stellenwert einnehmen sollte. Die Untersuchungsergebnisse zum Nachweis von Eimeria-Arten beim Kalb von 70 Sammelkotproben, hergestellt aus 10 Einzelkotproben (SKP10), bzw. von 30 Sammelkotproben, zusammengesetzt aus 5 Einzelkotproben (SKP5), wurden mit denen der zugehörigen Einzelkotproben (EKP) verglichen. Die Resultate der EKP (arithmetischer Mittelwert) und der zugehörigen SKP weisen mit den signifikant häufigeren Abweichungen im Bereich von bis zu 100 Oozysten pro Gramm Kot (OpG) eine geringe Differenz zwischen den beiden Verfahren auf. Durch den sicheren Nachweis von Eimeria-Oozysten bei einem erwarteten Oozystengehalt von nur 202 OpG (SKP10) und 122 OpG (SKP5) ist die Untersuchung von Kälbersammelkotproben, eine Methode mit geringem Arbeitsaufwand und geringen Untersuchungskosten, zum Nachweis einer klinischen oder subklinischen Kokzidiose geeignet. Bei 51 Pferdekotproben wurde jeweils dreimal das kombinierte Sedimentations-Flotations-Verfahren (KSFV), wobei die Entnahme von verschiedenen Lokalisationen der Kotprobe (aus der Randregion, dem Inneren oder aus beiden Lokalisationen) erfolgte, und jeweils dreimal das KSFV mit vorheriger Homogenisierung einer größeren Kotmenge zum Nachweis von Nematodeneier durchgeführt. Eine Anhäufung der Strongyliden- und Ascarideneier in einem bestimmten Bereich der Proben konnte durch die Untersuchungen der verschiedenen Lokalisationen (á 10 g Kot) nicht nachgewiesen werden, so dass eine weitgehend homogene Verteilung dieser Nematodeneier in einer Pferdekotprobe wahrscheinlich ist. Zudem konnten die Untersuchungsergebnisse des KSFV, bei welchem 10 g Kot untersucht werden, durch die vorherige Homogenisierung einer größeren Probenmenge nicht verbessert werden. Zum Nachweis von Nematoden beim Pferd sollte dem Labor eine ausreichende Probenmenge (ca. 50 g) zugesandt werden. Die Homogenisierung einer größeren Probenmenge vor der Durchführung einer diagnostischen Methode, bei der Aliquote von mindestens 10 g Kot Verwendung finden, ist unnötig. / The studies presented were carried out to compare different diagnostic methods for detection of protozoa and nematodes regarding sensitivity, expenditure of human labour and costs. Besides, the ability of the PCR for the molecular characterization of the Cryptosporidium spp. was tested exemplarily in faecal samples of hegdehogs. The examination of ninety faecal samples of suckling piglets showed a significantly higher sensitivity of faecal smears examined by autofluorescence microscopy (AM) compared to the flotation method (FV) using NaCl-sucrose solution and the combined sedimentation-flotation method (KSFV) using different flotation solutions (NaCl, ZnSO4, NaCl-sucrose) scanned by bright field microscopy. Moreover the expenditure of human labour by AM is considerably lower than FV and KSFV. The costs related to equipment for AM is justified in case of high sample throughput and by superior sensitivity. The enzyme immunoassay (EIA; ProSpecT® Cryptosporidium Microplate Assay) was the most sensitive method for diagnosis of cryptosporidiosis in calves (n = 103) compared to the carbol fuchsin (CF; HEINE 1981) and modified Ziehl-Neelsen (MZN; HENRIKSEN a. POHLENZ 1982) staining techniques. The sensitivity of the EIA was significantly higher than the MZN, if ten fields of view were scanned. 74 faecal samples of hedgehogs were examined with the EIA (ProSpecT®), an immunochromatographic method (FASTest® CRYPTO Strip), the MZN (HENRIKSEN u. POHLENZ (1981)) and a direct immunofluorescent assay (IFA; MERIFLUOR Cryptosporidium/Giardia). Cryptosporidium sp. were detected in 9 (EIA), 10 (FASTest®), 11 (MZN) und 12 (IFA) faecal samples. The hands on time of the FASTest® and CF is comparable to EIA while the IFA and MZN are more time-consuming. The examination of the FASTest®, IFA and EIA is combined with higher costs than the staining techniques, but they can be integrated in the work flow of a routine diagnostic laboratory easily and evaluation is simple. Moreover 45 faecal samples stored up to 27 days at different temperature (+6 °C, +16 °C, +30 °C, +40 °C) were examined to evaluate the influence of temperature on the results of EIA, CF and MZN. While the number of the positive samples of stained smears decreased in a temperature and time-dependent manner, the results of the EIA were not influenced by sample storage at any temperature, so that samples transported without cooling should be examined preferably by EIA. Nevertheless the CF due to its simplicity and low costs is suited for scanning of a high number of samples, if they were cooled continuously and examined within three days. The FASTest® is qualified for use in veterinary practice and stables, because the examination requires no microscope and the results are obtained immediately. The IFA, which can detect Crypotsporidium oocysts as well as Giardia cysts, is suited especially for faecal samples suspected to contain both protozoa. Cryptosporidial infections are very frequent in hedgehogs which are admitted for hibernation to hedgehog rehabilitation centres because of their insufficient body weight and weakness. Cryptosporidium spp. were detected in 29.8 % of 188 faecal samples of hedgehogs. The genotyping of Cryptosporidium spp. by PCR and RFLP-PCR based on the 18S ribosomal RNA gene were performed on 15 faecal samples of hedgehogs positive for Cryptosporidium spp. and suggested the presence of C. parvum in all samples. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA gene, actine gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new proposed as VIIa subtype family. Some of the Cryptosporidium oocysts excreted from hedgehogs are zoonotical (IIa subtype family) or anthropozoonotic(IIc subtype family). Thus hygienic measurements to avoid transmission are essential in hedgehog rehabilitation centres. The results of examination of 70 pooled faecal samples originating from 10 calves (SKP10) and 30 pooled faecal samples originating from 5 calves (SKP5) for detection of Eimeria spp. were compared with the arithmetic means of opg (oocysts per gram of faeces) counts of the respective single 10 or 5 samples. A low difference between both methods of less than 100 opg was significantly more frequently observed than higher differences. Low values of 202 opg and 122 opg were reliably detected in SKP10 und SKP5, respectively, and thus examination of pooled faecal samples appears to be suitably sensitive and cost effective to detect clinical and subclinical coccidiosis in calves. 51 faecal samples of horses were examined three times by KSFV for nematode eggs by taking aliquots from different locations of the same faecal samples (from the margin, from inside and from both locations). Thereafter the KSFV with the homogenisation of a larger amount of faeces was also carried out three times. The examination of samples from the different locations (each 10 g of faeces) delivered no evidence for accumulation of nematode eggs (strongyles and Parascaris equorum) in the faeces and thus the distribution of the nematode eggs appears sufficiently homogeneous in faecal samples of horses. Homogenisation of a larger amount of faeces did not improve the results of coproscopy. For diagnostic purposes 50 g faeces per sample should be shipped to the laboratory. The homogenisation of a larger amount of faeces before using a diagnostic method is dispensable, if aliquots of 10 g faeces are examined.

In dieser retrospektiven klinischen Studie wurden die Akten von 89 Patientinnen mit Ullrich-Turner-Syndrom ausgewertet, die zwischen 1974 und 2004 in der Universitätsklinik und Poliklinik für Kinder und Jugendliche in Leipzig behandelt worden sind. Berücksichtigt wurde die Verteilung der Karyotypen im Patientenkollektiv sowie das Auftreten von assoziierten Begleiterkrankungen. Das Alter bei Diagnosestellung, die Größe bei Diagnosestellung und die Gründe für die Verdachtsdiagnose Ullrich-Turner-Syndrom wurden analysiert. Darüber hinaus untersuchte der Autor die durchgeführten Maßnahmen zur Pubertätsinduktion im Hinblick auf ihren Beginn und Erfolg sowie deren Einfluss auf die Wachstumraten. Gut 50% der Patientinnen besaßen den Karyotyp 45,X, die anderen Karyotypen setzten sich aus Mosaiken zusammen. Assoziierte Begleiterkrankungen waren im Patientenkollektiv unterrepräsentiert. Das durchschnittliche Alter bei Diagnosestellung betrug 8,21 Jahre, es fiel während des Beobachtungszeitraumes signifikant ab. Der durchschnittliche Größen-SDS zum Zeitpunkt der Diagnosestellung betrug -2,86. Es wurde, verglichen mit den Empfehlungen der Leitlinien, eine verspätete Diagnosestellung konstatiert. Die Pubertätsinduktion begann mit durchschnittlich 13,93 Jahren mit einer signifikanten Reduktion im Verlauf. Die Dauer vom Beginn der Pubertätsinduktion bis zum Eintreten der Menarche betrug 2,51 Jahre, die Dauer vom Tannerstadium B2 zum Stadium B5 betrug gut 27 Monate. Eine Menarche wurde bei nur 65% der Patientinnen sicher beobachtet. Die Ergebnisse wurden kritisch überprüft und in den Kontext anderer Studien eingeordnet. Aus den Ergebnissen wurde gefolgert, dass das Ullrich-Turner-Syndrom, trotz einer positiven Entwicklung in den letzten Jahren, noch immer zu spät diagnostiziert wurde. Die Pubertätsinduktion verlief trotz der verzögerten Diagnosestellung hinsichtlich der Entwicklung der Tannerstadien erfolgreich. Demgegenüber blieb die Induktion der Menarche nur mäßig erfolgreich. Eine Beeinflussung der Wachstumraten durch die Östrogentherapie wurde nicht beobachtet.

In den westlichen Industrieländern ist Brustkrebs die häufigste krebsbedingte Todesursache bei Frauen. Bei der Diagnose von Brustkrebs ist für bestimmte Fragestellungen die Positronen-Emissions-Tomographie (PET) eine wichtige Ergänzung zur Mammographie und Sonographie. Die derzeit eingesetzten PET-Tracer 2-[18F]Fluor-2-desoxy-D-Glucose (FDG) und 16a-[18F]Fluor-17b-estra--diol (FES) können Mammakarzinome bzw. deren Metastasen nicht mit 100%iger Sicherheit nachweisen. Aus diesem Grund wurde eine [18F]-markierte Verbin-dung entwickelt, die möglicherweise die diagnostische Sicherheit bei Brustkrebs erhöhen kann: [18F]16a-Fluorestradiol-3,17b-disulfamat ([18F]FESDS). [18F]FESDS sollt dabei an Estronsulfatase binden, die in Mammakarzinomen signifikant höher exprimiert wird. Es wurde geprüft, ob [18F]FESDS für die Positronen-Emissions-Tomographie eignet ist. Die Ergebnisse zeigten, dass die Estronsulfatase nicht das alleinige Target von [18F]FESDS darstellt. Auch die Carboanhydrase hat eine sehr hohe Affinität zu dem potentiellen Tracer. Durch selektive Hemmung der Carboanhydrase mit Azetazolamid konnte jedoch in vitro die Estronsulfatase dargestellt werden. In vivo ist es nicht gelungen, mit [18F]FESDS die Estronsulfatase bei vorliegender Carboanhydrase-Hemmung besser darzustellen. Die Daten deuten darauf hin, dass eine Darstellung der Estronsulfatase mit der PET weder mit dem Radiotracer [18F]FESDS allein noch bei Koinjektion von [18F]FESDS mit Azetazolamid (zur CA-Hemmung) möglich ist.

Η παρούσα διπλωματική εργασία βρίσκεται στον ευρύτερο χώρο της μαθηματικής στατιστικής θεωρίας των γραφημάτων. Κύριος στόχος μας, όπως αναφέρει και ο τίτλος, είναι η μοντελοποίηση γραφημάτων, με απώτερο σκοπό την προσαρμογή, προσομοίωση και διάγνωση αυτών μέσω μοντέλων εκθετικών τυχαίων γραφημάτων. Το πρώτο κεφάλαιο δίνει μια συνοπτική παρουσίαση της διατύπωσης του προβλήματος και της θεωρίας των μοντέλων των εκθετικών τυχαίων γραφημάτων. Η βασική ιδέα είναι να θεωρήσουμε ως τυχαίες μεταβλητές τους δυνατούς δεσμούς μεταξύ των κόμβων ενός δοθέντος γραφήματος. Η γενική μορφή ενός μοντέλου εκθετικά τυχαίου γραφήματος καθορίζεται από κάποιες υποθέσεις σχετικές με τις εξαρτήσεις μεταξύ αυτών των τυχαίων μεταβλητών. Παρουσιάζουμε κάποιες διαφορετικές υποθέσεις εξάρτησης και τα αντίστοιχα μοντέλα, όπως τα γραφημάτα Bernoulli, τα δυαδικώς - ανεξάρτητα και τα τυχαία γραφήματα Markov. Επίσης, εξετάζουμε την ενσωμάτωση των χαρακτηριστικών, που μπορούν να έχουν οι κόμβοι, σε μοντέλα κοινωνικής επιλογής, δηλαδή, σε περιπτώσεις που οι συνδέσεις του γραφήματος μπορούν να προβλέψουν τα χαρακτηριστικά των κόμβων. Συνοψίζουμε κάποιες καινούργιες υποθέσεις εξάρτησης, που είναι πολυπλοκότερες των πρώτων τέτοιων υποθέσεων της σχετικής βιβλιογραφίας. Συζητούμε τις διαδικασίες της στατιστικής εκτίμησης, συμπεριλαμβανομένων των νέων μεθόδων για την εκτίμηση της μέγιστης πιθανοφάνειας Monte Carlo. Τέλος, παρουσιάζουμε τις νέες προδιαγραφές για μοντέλα εκθετικών τυχαίων γραφημάτων, που έχουν προτείνει οι Snijders et al., οι οποίες βελτιώνουν σημαντικά τα αποτελέσματα της προσαρμογής εμπειρικών δεδομένων για εκθετικά μοντέλα ομοιογενών τυχαίων γραφημάτων Markov. Επιπλέον, οι νέες αυτές προδιαγραφές μας βοηθούν να αποφύγουμε το πρόβλημα του σχεδόν-εκφυλισμού, που συχνά παρεμβάλλεται στη διαδικασία της προσαρμογής μοντέλων εκθετικών τυχαίων γραφημάτων Markov, ιδιαίτερα όταν αυτά προέρχονται από εμπειρικά δεδομένα, που έχουν υψηλό βαθμό μεταβατικότητας. Η μελέτη μιας τέτοιας νέας στατιστικής με υψηλότερης τάξης μεταβατικότητα επιτρέπει την εκτίμηση των παραμέτρων των μοντέλων των εκθετικών γραφημάτων σε πολλές (αλλά όχι όλες) περιπτώσεις, στις οποίες διαφορετικά θα ήταν αδύνατο να εκτιμηθούν οι παράμετροι των μοντέλων των ομοιογενών γραφημάτων Markov. Στο δεύτερο, τρίτο και τέταρτο κεφάλαιο της εργασίας εφαρμόζουμε τις παραπάνω μεθόδους, αντιστοίχως, για τρείς αναλύσεις εμπειρικών δεδομένων: το δίκτυο Florentine, το δίκτυο Faux Magnolia High και τα δίκτυα IPRED και SWPAT. Σε αυτά τα κεφάλαια, παρουσιάζουμε τις διαδικασίες της προσαρμογής, προσομοίωσης και διάγνωσης με παράθεση των αντίστοιχων εντολών, χρησιμοποιώντας τα πακέτα statnet - ermg και sna, τα οποία δουλεύουν στο περιβάλλον του πακέτου ελεύθερου λογισμικού R. Τέλος, στο παράρτημα της εργασίας δίνουμε μια σύντομη εισαγωγή στο περιβάλλον R και σε κάποιες γενικές εντολές αυτού. / This specific project has to do with mathematical statistical graph theory. Our main target is to fit, simulate and diagnose models through exponential random graph models. In the first chapter we give a short presentation of the problem and the theory of exponential random graph models. The main idea is to consider each tie of a given network (graph) as a random variable. The general form of an exponential random graph model is defined from some relative assumptions that have to do with the dependence between those random variables. We present some different dependence assumptions and the corresponding models, such as Bernoulli graphs, dyadic-independent and Markov random graphs. We also examine the incorporation of the characteristics that a node may have in social networks. We also discuss the process of statistical estimation, including three new methods for the estimation of Monte Carlo maximum likelihood. Finally, we present new specifications for exponential random graph models, which Snijders et al. have proposed. These new specifications allow us to avoid the problem of degeneration. In the second, third and fourth chapter we apply the above methods in order to analyze Florentine network data, Faux Magnolia High data and IPred And Swpat data. In those chapters, we present the procedures of fit, simulate and diagnose exponential random graph models displaying the corresponding commands of statnet-ergm and sna packages that work in R. Finally we give a short introduction to R and to some relative commands.

Virtual manipulatives are employed by both preservice and inservice teachers to enhance the instructional effectiveness of physical manipulatives and related tools by addressing limitations of access, cost, and adaptability. While research into the use of emerging technologies continues, there are several variables to consider when measuring the effects of virtual manipulative use. Research design, sampling characteristics, and the type of manipulative used may influence achievement. Variables that may influence the effectiveness of virtual manipulatives include: previous experience with computers, grade level, mathematical topic, treatment length, student attitudes toward mathematics, and computer-to-student ratio.

Orientador: Marcelo Carvalho Minhoto Teixeira Filho / Resumo: A produção de alimento face a demanda mundial é um desafio para este século, devendo-se, obrigatoriamente, destinar atenção ao ganho de eficiência produtiva, incorporada as tecnologias disponíveis. Dentre as hortaliças folhosas, a alface se destaca em toda culinária global. A produção atual da alface tem passado por grandes avanços, atribuídos especialmente à técnica de produção hidropônica, contudo, ainda é necessita melhorias na adaptabilidade da tecnologia hidropônica às condições climáticas das diversas regiões e às diferentes necessidades da gama de cultivares presentes no mercado. Neste cenário, a proposição desta pesquisa surge em resposta à demanda de informações pelo setor produtivo, cumprindo seu papel na integração entre ensino, pesquisa e extensão rural, de maneira a contribuir modestamente com o setor produtivo. Para isso, foram realizados três experimentos isolados em sistema hidropônico NFT (Técnica de Fluxo Laminar de Nutrientes), com o objetivo de avaliar a nutrição e a produção econômica de alface americana em função da vazão, periodicidade de exposição e condutividade elétrica da solução nutritiva, respectivamente. O delineamento experimental foi em blocos casualisados, com cinco repetições e cada tratamento foi composto por 15 plantas. No experimento 1, os tratamentos utilizados constaram da aplicação de solução nutritiva nas vazões de 0,5; 1; 2 e 4 litros por minuto em cada canal de cultivo. No experimento 2, os tratamentos foram quatro combinações de int... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor