Tagetes minuta L. is a weedy herb that has been a rich source of fragrant oils, used as in the perfume and flavour industry. T. minuta achenes germinate erratically under field conditions. However, at the optimal germination temperature of 25 °C, 100 % germination is attained within 48 h of imbibition. The achenes are thermoinhibited at 35 °C. The aims of this project were to assess the role of cytokinins (CKs) in normal germination at 25 °C, and to investigate the factors that regulate thermoinhibition at 35 °C. CKs were extracted from achenes germinating at 25 °C at 0, 24; 48; 96 and 144 h after imbibition. Two different purification techniques were used, namely Dowex cation exchange resin followed by paper chromatography, or high performance liquid chromatography (HPLC). CK-like activity was tested with the soybean callus bioassay. With both techniques, a peak in CK-like activity appeared 24 h after imbibition, which coincides with the period during which most of the achenes germinated. For quantitative analysis, HPLC\mass spectrometry (MS) techniques were used. The isoprenoid CKs were far more abundant in T. minuta achenes than the aromatic CKs. cis-Zeatin (cZ) and its derivatives were the most abundant CKs. In total, 19 CK compounds were detected, including 4 free bases and a number of corresponding conjugates. Benzyladenine (BA) was the only aromatic CK detected. There was no common time at which active free base maximal concentrations were detected, suggesting that different CKs may have specific roles in the germination process, and thus peak at different times. This in turn suggests that germination is not a single process, but rather a correlative process involving a number of events, with specific CKs having specific roles relating to these correlative events. There is sufficient evidence obtained from both the soybean callus bioassay and HPLC/MS analysis to suggest that CKs have an active role in T. minuta germination. A decline in free BA during germination without corresponding conjugation, suggests that BA is actively used in early germination processes, possibly in the stimulation of DNA synthesis. Secondly, there was a distinct dihydrozeatin (DHZ) peak obtained at 24 h. Roughly 75 % of the achenes germinate between 16 and 26 h, thus it is likely that DHZ has an active role during the germination of T. minuta. Although CKs are probably not involved in the breaking of dormancy per se, the distinct peak in CK-like activity obtained in the bioassays, 24 h after imbibition, suggests that CKs have an active role in the germination of T. minuta. With respect to the regulation of thermoinhibition, a number of exogenous treatments were applied, including hormones [gibberellins (GA₄₊₇), abscisic acid (ABA), ethylene and a number of CKs], adenosine triphosphate (ATP) and incubation in 100 % oxygen. ABA was extracted from thermoinhibited and germinating achenes to assess the role of ABA in thermoinhibition and germination. While exogenous 0.1 mg L¯¹ GA₄₊₇ application slightly improved normal germination at 25°C, no treatments were effective in alleviating thermoinhbibition in T. minuta achenes. Thermoinhibition in T. minuta achenes may be under hormonal regulation, as there is strong evidence for the role of ABA in the maintenance of dormancy and thermoinhbition. High ABA levels were found in dry control samples. Additionally, exogenous ABA application inhibited normal germination, and the commencement of germination was accompanied by a decrease in endogenous ABA levels. A number of experiments relating to the imposition of thermoinhibition were carried out. Thermoinhibition appears to be very rapidly imposed. Germination is rapidly inhibited following shifting to higher thermoinhibitory temperatures, even after prolonged exposure to optimal germination temperatures. Results suggest active de novo biosynthesis of ABA in thermoinhibited achenes. Active biosynthesis of ABA during thermoinhibition suggests that this phytohormone is essential in the maintenance of thermoinhibition of T. minuta achenes. It thus appears that ABA is synthesized in the achenes in response to elevated temperatures that are unfavourable for germination to proceed. Unfavourable environmental conditions result in an achene-mediated inhibition of germination, which appears to be initiated and maintained by elevated levels of endogenous ABA. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.

When imbibed at their optimum germination temperature of 25°C, achenes of Tagetes minuta L. germinate over a period of approximately 48 h. At temperatures of between 35°C and 39°C, the achenes do not germinate but enter into a state of thermoinhibition. These supra-optimal conditions do not harm the achenes, however, and when the temperature is reduced below 35°C radicle emergence may be observed within 4 h. Achenes which have been thermoinhibited for periods of 24 h or more show "accelerated germination" which takes only 24 h, although the actual germination curve is identical to that of normally germinated achenes. This suggests that the achenes are metabolically active at thermoinhibitory temperatures and undergo most of the processes of normal germination, but that at some point any further development is halted, preventing radicle emergence. When the temperature is reduced, this block on germination is removed and since the achenes are already primed for germination, this occurs within a short time. An analysis of the proteins produced by germinating and thermoinhibited achenes was conducted using SDS-polyacrylamide gel electrophoresis (SDS-PAGE). This procedure was able to resolve approximately 40 different protein bands, but no differences were observed between thermoinhibited and germinating achenes. Two dimensional polyacrylamide gel electrophoresis (20-PAGE) was able to resolve approximately 200 individual polypeptides. The vast majority of polypeptides in T. minuta achenes are acidic, although the number of neutral to basic polypeptides increases as germination progresses. Ten polypeptides were identified which were specific to thermoinhibited achenes. These formed two distinct groups on the twodimensional gels. The larger group contained seven proteins, ranging in size from 22 kDa to 26.7 kDa and with isoelectric points of between 3.0 and 4.0. The smaller group contained three polypeptides with molecular weights of about 14 kDA and a pi of approximately 3.0. These polypeptides were all extremely specific to thermoinhibited achenes and declined rapidly when the incubation temperature was reduced, in a manner which correlated with an increase in the germinability of the achenes. Several characteristics of the expression of these polypeptides were similar to characteristics of embryo-dormancy in seeds where dormancy is thought to be actively imposed by the expression of specific dormancy-associated genes. This, along with the very tightly-regulated nature of these 10 polypeptides, suggests that thermoinhibition in T. minuta may be regulated through gene expression and that these ten polypeptides may represent the products of genes responsible for preventing radicle emergence at unfavourable temperatures. Since these polypeptides were only resolved using silver-staining and could not therefore be used for amino acid sequence analysis, this hypothesis was further investigated using differential display of mRNA to isolate genes which are expressed specifically in thermoinhibited achenes. A large number of cDNA fragments which were specific to either germinating or thermoinhibited achenes were identified and extracted from the differential display gels. Those cDNAs specific to the thermoinhibited achenes were taken for further analysis. Of the 62 fragments excised from the gels, 25 could be reamplified to generate single bands of the correct size on agarose gels. A further 22 cDNAs produced multiple bands, where one band was much brighter than the others and correlated with the size of the original fragment. Thirteen of the cDNAs which' generated single bands were cloned into the plasmid vector pGEM®-T Easy and transformed into either Escherichia coli JM109 or E. coli XL1-Blue. Recombinant colonies were identified using blue-white colour selection and the presence of the insert confirmed by colony blotting and restriction analysis. Three clones were chosen for each of the cDNAs. Reverse northern analysis confirmed that all 39 clones were specific to the mRNA pool of thermoinhibited achenes. High quality sequence data were obtained for 27 of the cDNA samples, the remainder appeared to have been degraded in transit. Alignment of the various sequences revealed that a total of 14 different sequences had been cloned, indicating that several of the bands isolated from the differential display gels contained multiple sequences. Electronic homology searches tentatively identified three of the sequences, whilst the remainder did not show significant homology to any known sequences. Of the cDNAs identified in this way, one may encode a plant transcription factor-like or nuclear RNA-binding protein whilst the other two may encode an RNase-L Inhibitor-like protein and a miraculin homologue. The potential roles of such genes in the imposition or maintenance of the thermoinhibited state are discussed. Although further research needs to be conducted to isolate full length cDNA sequences and to determine their exact expression patterns in germinating and thermoinhibited achenes, these results are consistent with the hypothesis that thermoinhibition in T. minuta achenes is under positive genetic control in a manner analogous to embryo dormancy. This thesis represents the first molecular study of thermoinhibition as well as the first report of active control over this phenomenon in any species. Since thermoinhibition, unlike dormancy, can be rapidly imposed and released under strictly controlled conditions without the need for any dormancy breaking treatment, T. minuta achenes represent an excellent model system for studies on the molecular control of seed germination. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2003.

Made available in DSpace on 2014-08-20T14:38:03Z (GMT). No. of bitstreams: 1 dissertacao_diane_bender_schiavon.pdf: 500386 bytes, checksum: 48fa2cd250b94f5f938cb28a34fe2107 (MD5) Previous issue date: 2011-02-24 / Bovine Mastitis is a disease that causes high damage to milk production. Phytotherapy is the basis of Tagetes minuta L. can be used for disease control agroecological production units. The study aimed to evaluate application of antiseptics from medicinal plant from the postmilking teat disinfection. The treatments consisted of commercial iodine with treatment 1, used as teat pos-dipping on the right side, and hydro alcoholic extract from leaves of Tagetes minuta and macerated seeds Linum usitatissimum L., both at 10%, used on teats left as pos-dipping, cows in a commercial production unit that had, in average of 60 milking animals during the experiment that lasted 12 weeks. The prevalence of subclinical mastitis was evaluated using the CMT test once a week. Test mug of dark background was done at all milking. Sample of milk from all quarters was collected weekly for the isolation and characterization of present microorganisms . The incidence of new intramammary infections was calculated using the number of infection-free days by quarters, counted by weekly analysis of milk from all quarters in blood agar culture, as the denominator and the number of new cases as the numerator, defined as culture positive by Staphylococcus spp. or Streptococcus spp. in Sampling of milk per room. The index was corrected to 1000 / 4 days. Exclusion criteria: infection with Streptococcus spp or Staphylococcus spp in early labor and up to two weeks after a collection microbiologically positive. Animals with two successive collections negative after a positive were considered cured and reintroduced in the counting of days without infection. The chi-square test was used to compare the frequency of events between treatments. The CMT's weekly prevalence ranged in group 1 between 29.5% and 17.1%, and group 2 from 29.7% to 19.6%, not significantly different in any of weeks. The incidence of positive cultures for Staphylococcus / Streptococcus was 3.93 and 6.96 / 1000 / 4 / day for Groups 1 and 2 respectively, p = 0.057. There were four cases of mastitis clinic during the experiment, two in each treatment. We concluded the use of plant extracts for the disinfection of teats pos-dipping can be useful systems for milk production agro ecological. / Mastite bovina é a doença que causa os maiores prejuízos na produção leiteira. A fitoterapia a base de Tagetes minuta pode ser utilizada para controle da enfermidade em unidades de produção agroecológicas. O trabalho objetivou avaliar a aplicação de anti-sépticos obtidos de planta medicinal na desinfecção de tetos pós-ordenha. Os tratamentos consistiram em iodo comercial como tratamento 1, utilizado como pós-dipping nos tetos do lado direito, e extrato hidroalcoolico de folhas de Tagetes minuta L. e macerado de sementes de Linum usitatissimum L., ambos à 10%, utilizados nos tetos esquerdos como pos-dipping de vacas em uma unidade de produção comercial que possuía, em média, 60 animais em ordenha durante o experimento que teve duração de 12 semanas. A prevalência de mastite sub-clínica foi avaliada utilizando o teste de CMT uma vez por semana. Teste da caneca de fundo escuro foi realizado em todas as ordenhas. Amostra de leite de todos os quartos foi coletada semanalmente para isolamento e caracterização dos microrganismos presentes. A incidência de novas infecções intra-mamárias foi calculada utilizando o número de dias livres de infecção por quarto, contados pela análise semanal de leite de todos os quartos em cultura de Agar sangue, como denominador, e o número de casos novos como numerador, definido como cultura positiva para Staphylococcus spp. ou Streptococcus spp. na amostragem do leite por quarto. O índice foi corrigido para 1000 quartos dia. Foram critérios de exclusão: infecção por Streptococcus spp ou Staphylococcus spp no início do trabalho e até duas semanas após uma coleta microbiologicamente positiva. Animais com duas coletas sucessivas negativas após uma positiva foram considerados curados e reintroduzidos na contagem de dias sem infecção. O teste do qui-quadrado foi utilizado para comparação das frequências dos eventos entre os tratamentos. A prevalência semanal do CMT variou no grupo 1 entre 29,5% e 17,1%, e no grupo 2 de 29,7% a 19,6%, não diferindo significativamente em nenhuma das semanas. A incidência de cultura positiva para Staphylococcus/Streptococcus foi de 3,93 e 6,96/1000 quartos/dia para os grupos 1 e 2 respectivamente, com p=0,057. Houve quatro casos de mastite clínica durante o experimento, dois em cada tratamento. Concluímos que o uso de extratos de plantas na desinfecção de tetos pós-ordenha pode ser útil aos sistemas de produção de leite agroecológico.

O fitoterápico Xarope de Agrião Composto Cibecol® é uma associação de extratos de Roripa nasturtium Rusby (Agrião), Musa spp. (Bananeira), Ficus carica Linné (Figueira), Tagetes minuta Linné (Chinchilia) e mel de abelhas. Avaliou-se a segurança deste fitoterápico através de estudos de toxicidade aguda e subcrônica, tendo como base a resolução Nº 90, de 16 de março de 2004 da ANVISA. Para o teste de toxicidade aguda, ratos Wistar de ambos os sexos foram tratados por via oral com uma única dose de 26 ml/kg, correspondendo a 20 vezes a dose terapêutica indicada pelo fabricante para seres humanos adultos. Os resultados revelaram não haver sinais de toxicidade sistêmica, não causando interferência no desenvolvimento ponderal dos animais, nos consumos de água e ração, nas produções de urina e fezes, bem como alterações macroscópicas nos órgãos dos animais. Avaliou-se também a exposição a doses repetidas do fitoterápico. Constituíram-se em quatro grupos experimentais (10 animais/sexo/dose), onde administrou-se por via oral a ratos Wistar, durante 30 dias, doses diárias de 1,3 ml/kg, 6,5 ml/kg e 13 ml/kg, respectivamente a dose terapêutica indicada pelo fabricante para seres humanos adultos, 5 vezes, e 10 vezes a dose terapêutica, além de um grupo controle, onde foi administrado o veículo do fitoterápico. Os resultados revelaram ausência de toxicidade sistêmica, fundamentados na ausência de alterações hematológicas e bioquímicas sangüíneas, bem como no peso e análises histopatológicas dos órgãos, nos diferentes grupos. Concluiu-se que, a utilização do fitoterápico nas doses e períodos referidos, pode ser considerada segura. / The phytotherapic Xarope de Agrião Composto Cibecol® is an association of extracts of Roripa nasturtium Rusby (water-cress), Musa spp. (banana tree), Ficus carica Linné (fig tree), Tagetes minuta Linné (Chinchilia) and honey. The safety of the phytotherapic was evaluated through studies of acute and sub-chronic toxicity, being based in the resolution Nº 90, March 16th, 2004 from ANVISA. For the test of acute toxicity Wistar rats of both sexes were treated orally with a single dose of 26 ml/kg, which corresponds to 20 times the therapeutic dose indicated by the producer for adult humans. The results revealed that there are no signals of systemic toxicity, no interference in the development of weight gain in the animals, in water and feed consume, in the production of urine and feces, neither macroscopic alterations in the animals’ organs. It was also evaluated the exposition of repeated doses of the phytotherapic (sub-chronic toxicity). Four experimental groups (10 animals/sex/dose) were orally treated during 30 days with daily doses of 1.3 ml/kg, 6.5 ml/kg and 13 ml/kg, respectively the therapeutic dose indicated to humans, 5 times, and 10 times the therapeutic dose, and a control group, receiving the phytotherapic vehicle. The results revealed the absence of systemic toxicity, based in the absence of hematological and blood biochemical alterations, as well as weigh and histopatological analysis of organs, in the different groups. It was concluded that the utilization of the referred phytotherapic in the mentioned doses and periods might be considered secure.

O fitoterápico Xarope de Agrião Composto Cibecol® é uma associação de extratos de Roripa nasturtium Rusby (Agrião), Musa spp. (Bananeira), Ficus carica Linné (Figueira), Tagetes minuta Linné (Chinchilia) e mel de abelhas. Avaliou-se a segurança deste fitoterápico através de estudos de toxicidade aguda e subcrônica, tendo como base a resolução Nº 90, de 16 de março de 2004 da ANVISA. Para o teste de toxicidade aguda, ratos Wistar de ambos os sexos foram tratados por via oral com uma única dose de 26 ml/kg, correspondendo a 20 vezes a dose terapêutica indicada pelo fabricante para seres humanos adultos. Os resultados revelaram não haver sinais de toxicidade sistêmica, não causando interferência no desenvolvimento ponderal dos animais, nos consumos de água e ração, nas produções de urina e fezes, bem como alterações macroscópicas nos órgãos dos animais. Avaliou-se também a exposição a doses repetidas do fitoterápico. Constituíram-se em quatro grupos experimentais (10 animais/sexo/dose), onde administrou-se por via oral a ratos Wistar, durante 30 dias, doses diárias de 1,3 ml/kg, 6,5 ml/kg e 13 ml/kg, respectivamente a dose terapêutica indicada pelo fabricante para seres humanos adultos, 5 vezes, e 10 vezes a dose terapêutica, além de um grupo controle, onde foi administrado o veículo do fitoterápico. Os resultados revelaram ausência de toxicidade sistêmica, fundamentados na ausência de alterações hematológicas e bioquímicas sangüíneas, bem como no peso e análises histopatológicas dos órgãos, nos diferentes grupos. Concluiu-se que, a utilização do fitoterápico nas doses e períodos referidos, pode ser considerada segura. / The phytotherapic Xarope de Agrião Composto Cibecol® is an association of extracts of Roripa nasturtium Rusby (water-cress), Musa spp. (banana tree), Ficus carica Linné (fig tree), Tagetes minuta Linné (Chinchilia) and honey. The safety of the phytotherapic was evaluated through studies of acute and sub-chronic toxicity, being based in the resolution Nº 90, March 16th, 2004 from ANVISA. For the test of acute toxicity Wistar rats of both sexes were treated orally with a single dose of 26 ml/kg, which corresponds to 20 times the therapeutic dose indicated by the producer for adult humans. The results revealed that there are no signals of systemic toxicity, no interference in the development of weight gain in the animals, in water and feed consume, in the production of urine and feces, neither macroscopic alterations in the animals’ organs. It was also evaluated the exposition of repeated doses of the phytotherapic (sub-chronic toxicity). Four experimental groups (10 animals/sex/dose) were orally treated during 30 days with daily doses of 1.3 ml/kg, 6.5 ml/kg and 13 ml/kg, respectively the therapeutic dose indicated to humans, 5 times, and 10 times the therapeutic dose, and a control group, receiving the phytotherapic vehicle. The results revealed the absence of systemic toxicity, based in the absence of hematological and blood biochemical alterations, as well as weigh and histopatological analysis of organs, in the different groups. It was concluded that the utilization of the referred phytotherapic in the mentioned doses and periods might be considered secure.

O fitoterápico Xarope de Agrião Composto Cibecol® é uma associação de extratos de Roripa nasturtium Rusby (Agrião), Musa spp. (Bananeira), Ficus carica Linné (Figueira), Tagetes minuta Linné (Chinchilia) e mel de abelhas. Avaliou-se a segurança deste fitoterápico através de estudos de toxicidade aguda e subcrônica, tendo como base a resolução Nº 90, de 16 de março de 2004 da ANVISA. Para o teste de toxicidade aguda, ratos Wistar de ambos os sexos foram tratados por via oral com uma única dose de 26 ml/kg, correspondendo a 20 vezes a dose terapêutica indicada pelo fabricante para seres humanos adultos. Os resultados revelaram não haver sinais de toxicidade sistêmica, não causando interferência no desenvolvimento ponderal dos animais, nos consumos de água e ração, nas produções de urina e fezes, bem como alterações macroscópicas nos órgãos dos animais. Avaliou-se também a exposição a doses repetidas do fitoterápico. Constituíram-se em quatro grupos experimentais (10 animais/sexo/dose), onde administrou-se por via oral a ratos Wistar, durante 30 dias, doses diárias de 1,3 ml/kg, 6,5 ml/kg e 13 ml/kg, respectivamente a dose terapêutica indicada pelo fabricante para seres humanos adultos, 5 vezes, e 10 vezes a dose terapêutica, além de um grupo controle, onde foi administrado o veículo do fitoterápico. Os resultados revelaram ausência de toxicidade sistêmica, fundamentados na ausência de alterações hematológicas e bioquímicas sangüíneas, bem como no peso e análises histopatológicas dos órgãos, nos diferentes grupos. Concluiu-se que, a utilização do fitoterápico nas doses e períodos referidos, pode ser considerada segura. / The phytotherapic Xarope de Agrião Composto Cibecol® is an association of extracts of Roripa nasturtium Rusby (water-cress), Musa spp. (banana tree), Ficus carica Linné (fig tree), Tagetes minuta Linné (Chinchilia) and honey. The safety of the phytotherapic was evaluated through studies of acute and sub-chronic toxicity, being based in the resolution Nº 90, March 16th, 2004 from ANVISA. For the test of acute toxicity Wistar rats of both sexes were treated orally with a single dose of 26 ml/kg, which corresponds to 20 times the therapeutic dose indicated by the producer for adult humans. The results revealed that there are no signals of systemic toxicity, no interference in the development of weight gain in the animals, in water and feed consume, in the production of urine and feces, neither macroscopic alterations in the animals’ organs. It was also evaluated the exposition of repeated doses of the phytotherapic (sub-chronic toxicity). Four experimental groups (10 animals/sex/dose) were orally treated during 30 days with daily doses of 1.3 ml/kg, 6.5 ml/kg and 13 ml/kg, respectively the therapeutic dose indicated to humans, 5 times, and 10 times the therapeutic dose, and a control group, receiving the phytotherapic vehicle. The results revealed the absence of systemic toxicity, based in the absence of hematological and blood biochemical alterations, as well as weigh and histopatological analysis of organs, in the different groups. It was concluded that the utilization of the referred phytotherapic in the mentioned doses and periods might be considered secure.

En esta tesis se estudió el fraccionamiento de aceites esenciales obtenidos de plantas aromáticas comunes en el país mediante el uso de dióxido de carbono supercrítico, con el objetivo de obtener compuestos o fracciones con actividad biocida. Se seleccionaron tres plantas: Tagetes minuta, Salvia officinalis y Mentha piperita. La extracción del aceite esencial se realizó mediante hidrodestilación, determinándose su composición mediante cromatografía de gases. Como parte de esta tesis, se construyó un equipo experimental de alta presión, con el cual se realizaron mediciones de solubilidad y composición de la fase supercrítica para cada sistema [CO2 + aceite esencial] en distintas condiciones de presión y temperatura (70–120 bar, 313–323 K), utilizando el método dinámico o de gas saturado. Se realizaron también ensayos de fraccionamiento semicontinuo para determinar la influencia de la composición en el grado de separación. Con los resultados experimentales obtenidos se validó un modelo termodinámico predictivo basado en la ecuación de estado a contribución grupal (GC-EOS), previo ajuste de los parámetros necesarios con datos provenientes de otras fuentes. Utilizando dicho modelo como base, y reduciendo la complejidad de los sistemas en términos de dos o tres pseudo-componentes, se realizaron simulaciones de columnas de fraccionamiento de múltiples etapas en contra-corriente, analizando la influencia de las variables operativas (presión, temperatura, relación solvente/ alimentación, relación de reflujo, etc.) con el objetivo de hallar condiciones recomendables de operación para separar los compuestos biocidas de interés. En el análisis, se utilizaron herramientas y consideraciones de la Ingeniería del Equilibrio de Fases. En el caso del aceite de T. minuta se procuró separar la fracción de cetonas insaturadas (ocimenonas); en el caso del aceite de S. officinalis se planteó la purificación de la fracción de monoterpenos oxigenados, separándolos a la vez de los hidrocarburos y de los sesquiterpenos; en el caso del aceite de M. piperita se planteó la separación de mentona y mentol. La actividad biocida de las fracciones obtenidas experimentalmente a partir del aceite de T. minuta fueron ensayadas frente a dos parásitos: el gorgojo del maíz y el nemátodo del nudo de la raíz. / The fractionation of essential oils from local aromatic species using supercritical carbon dioxide was studied, in order to obtain compounds or fractions with biocidal activity. Three species were chosen: Tagetes minuta, Salvia officinalis y Mentha piperita. The essential oil was extracted by hydrodistillation and its composition determined by gas chromatography. As a part of this project, a lab-scale high-pressure experimental apparatus was built. With this equipment, measurements of solubility and composition in the supercritical phase were performed for each [CO2 + essential oil] system at different pressure and temperature conditions (70–120 bar, 313–323 K), using a dynamic or gas saturated method. Semi-continuous fractionation experiments were also carried out in order to determine the effect of composition in the separation performance. With the experimental data a predictive thermodynamic model based on the group contribution equation of state (GC-EOS) was validated, after fitting the necessary parameters with data from other sources. With this model as background, and reducing the systems complexity in terms of two or three pseudo-components, simulations of multistage counter-current columns were carried out, analyzing the influence of the operation parameters (pressure, temperature, solvent-to-feed ratio, reflux ratio, etc.), in order to find recommended conditions for the separation of the biocidal components. In this analysis, phase equilibrium engineering tools and considerations were applied. In the case of T. minuta, the goal was to isolate the unsaturated ketones fraction (ocimenones); in the case of S. officinalis the goal was the separation of the oxygenated monoterpenes fraction from both the hydrocarbon and the sesquiterpene fraction; finally in the case of M. piperita the separation between menthone and menthol was proposed. The biocidal activity of the fractions obtained experimentally from T. minuta oil was tested against the corn weevil and the root knot nemathod.

Made available in DSpace on 2016-01-26T12:51:23Z (GMT). No. of bitstreams: 1 waldemirpereiralima_tese.pdf: 266828 bytes, checksum: 4d3bfe97aba0900f9364737e7267f696 (MD5) Previous issue date: 2010-02-18 / Tropical diseases have been a phenomenon of global concern, and among them the Dengue, because the mosquito that transmits the virus, Aedes aegypti (Diptera: Culicidae), became out of control, and new outbreaks have been discovered each day. Several means have been used in an attempt to control the insect life cycle. However, the indiscriminate use of many agents took the insect to develop some resistance to them. New perspectives are emerging as we find in nature means to control these mosquitoes. Is the case of essential oil of Tagetes minuta L. (Asteraceae) which, according to recent studies, plays an important role in the control of insects, including the Aedes aegypti. Objectives: develop a procedure for feeding female Aedes aegypti that does not cause stress in Swiss Balb/C mice, evaluate the toxicity and residual effect of essential oil of Tagetes minuta L (Asteraceae) on populations of Aedes aegypti. Methods: Swiss Balb/C mice were anesthetized arranged in the cage for feeding female mosquitoes for a period of one hour, three times a week. The essential oil was used in concentrations of 0,2 mL L-1; 0,225 mL L-1; 0,25 mL L-1; 0,275 mL L-1 and 0,3 mL L-1 starting from an initial dilution of 200mL L-1 in acetone, in one hundred third-stage larvae coming from Bauru-SP, São José do Rio Preto-SP, respectively, sensitive and resistant to temephos, and one hundred larvae of Rockefeller-USA. The tests were performed in four times, with three replicates for three consecutive days. Results: There was no pain or stress observed in mice during blood feeding by Ae aegypti. There was no difference between populations for susceptibility to Tagetes minuta L. and tests showed LC50 of 0,24; 0,25 and 0,21 mL L-1 and CL99, 9 at 0,35; 0,39 and 0,42 mL L- 1, respectively, for Rockefeller, Bauru and São José do Rio Preto. There was no residual effect of the oil. Conclusions: Sedation of Swiss Balb/C mice has provided it serve as a food source for female Aedes aegypti without pain or stress, and can be used by other researchers, since it was approved by the Ethics Committee of Animal Experiments (CEEA) of this institution. The LC50 of Tagetes minuta L. were equal, respectively, for larvae of the Rockefeller strain, Bauru, São José do Rio Preto, and the CL99, 9 were equal, respectively, for larvae of the Rockefeller strain, Bauru, São José do Rio Preto. The resistance of larvae to temephos had no effect on lethal action exerted by the essential oil of Tagetes minuta L. Tests for evaluation of a residual oil after a week of contact, had zero mortality. The essential oil of Tagetes minuta L can be used as an alternative larvicide against Aedes aegytpi, however new studies are needed in order to isolate the fraction of larvicidal oil and incorporate other ingredients to increase their stability so that the product can have the residual effect. / As doenças tropicais têm sido um fenômeno de preocupação mundial, e entre elas a dengue, uma vez que o mosquito transmissor dos vírus, Aedes aegypti (Diptera: Culicidae), tornou-se fora de controle, e novos focos têm sido descobertos a cada dia. Vários meios já foram utilizados na tentativa de controlar o ciclo de vida inseto. Entretanto, o uso indiscriminado de muitos agentes levou o inseto a desenvolver certa resistência aos mesmos. Novas perspectivas estão surgindo já que podemos encontrar na própria natureza meios de controlar esses Culicideos. É o caso do óleo essencial de Tagetes minuta L. (Asteraceae) que, de acordo com estudos recentes, desempenha importante papel no controle de insetos, inclusive do Aedes aegypti. Objetivos: Desenvolver um procedimento de alimentação de fêmeas de Aedes aegypti que não cause estresse em camundongo Swiss Balb/C; avaliar a toxicidade e o efeito residual do óleo essencial de Tagetes minuta L (Asteraceae) em populações de Aedes aegypti. Material e métodos: Camundongos Swiss Balb/C anestesiados e sedados foram dispostos em gaiola para alimentação de fêmeas do mosquito por um período de uma hora três vezes por semana. O óleo essencial, foi utilizado nas concentrações de 0,2mL L-1; 0,225mL L-1; 0,25mL L-1; 0,275mL L-1 e 0,3mL L-1 partindo-se de uma diluição inicial de 200mL L-1 em Acetona, em cem larvas de terceiro estágio procedentes de Bauru-SP e São José do Rio Preto-SP, respectivamente, sensíveis e resistentes ao temephos, e, cem larvas de Rockefeller-EUA. Os testes foram realizados em quadriplicata, com três repetições, por três dias consecutivos. Resultados: Não foi observado dor ou estresse dos camundongos durante hematofagia pelas fêmeas de Ae aegypti. Não houve diferença entre as populações quanto à susceptibilidade a Tagetes minuta e os ensaios demonstraram CL50 de 0,24, 0,25 e 0,21mL L-1 e CL99,9 em 0,35, 0,39 e 0,42mL L-1, respectivamente, para Rockfeller, Bauru e São José do Rio Preto. Não foi observado efeito residual do óleo. Conclusões: A anestesia do camundongo Swiss Balb/C propiciou que este servisse como fonte alimentar para fêmeas de Aedes aegypti sem sofrer dor ou estresse, podendo este ensaio ser utilizado por como modelo, uma vez que foi aprovado pelo Comitê de Ética em Experimentação Animal (CEEA). As CL50 de Tagetes minuta L. foram iguais, respectivamente, para larvas da cepa Rockefeller, Bauru e São José do Rio Preto, assim como as CL99,9 que foram iguais, respectivamente, para larvas de da cepa Rockefeller, Bauru e São José do Rio Preto. A resistência das larvas do Culicideo ao temephos, não interferiu na ação letal exercida pelo óleo essencial de Tagetes minuta L. Os testes para avaliação do período residual do óleo após uma semana de contato, apresentaram mortalidade zero. O óleo essencial de Tagetes minuta L pode ser utilizado como larvicida alternativo contra Aedes aegytpi, entretanto novos estudos devem ser desenvolvidos com o objetivo de isolar a fração larvicida do óleo e incorporar outros componentes visando aumentar sua estabilidade, de maneira que o produto tenha efeito residual.

El exceso de metales pesados en suelos y agua es uno de los problemas actuales más relevantes por el riesgo de toxicidad que implica su ingreso a las cadenas tróficas. El uranio (U) es un constituyente natural de todos los suelos y la mayoría de las aguas naturales, pero se encuentra en mayor concentración en algunas rocas y minerales, que pueden ser explotados para obtener concentrados de U utilizables en la generación de energía. El uso potencial de plantas para la limpieza de suelos y aguas contaminados con niveles excesivos de Uranio derivados de su manipulación está en estudio, habiéndose llegado en un caso a una instalación de rizofiltración por girasol a escala piloto. Este trabajo tiene como objetivo general la contribución al conocimiento sobre la absorción de U en plantas terrestres y sobr el comportamiento de las mismas en medios contaminados con este metal pesado. Se ha subdividido en dos objetivos específicos: 1. Estudiar la capacidad potencial de absorción de U in situ de algunas especies vegetales de aparición espontánea en un área minera; 2. Estudiar el comportamiento y tolerancia de la remolacha roja (Beta vulgaris) a distintas concentraciones de U en condiciones de hidroponía en laboratorio. El coirón (Stipa sp) fue la única especie vegetal que se encontró en todos los suelos, incluyendo las escombreras de colas de mineral, tolerando concentraciones de U en el sustrato de hasta 280 mgU kg-1. Tagetes minuta (chin-chil o chil-chil) se encontró en sustratos de hasta 280 mgU kg-1. Las dos especies de jarilla, Larrea divaricata y Larrea nitida, en cambio, sólo estuvieron presentes en sustratos con baja contaminación. En todos los casos la acumulación de U y el factor de transferencia sustrato-planta o factor de bioacumulación (FB) fueron bajos. En el cultivo hidropónico, la remolacha sobrevivió en solución nutritiva con distintas concentraciones de U agregado entre 0 y 5 mgU L-1 durante 15 días. La tolerancia al U estaría asociada a la presencia de P, e / López, SC. (2009). Estrés por uranio en plantas. Acumulación en especies vegetales de zonas mineras y estudio de respuestas fisiológicas en remolacha (Beta vulgaris) [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/5764 / Palancia