ABSTRACT GENETIC VARIABILITY IN HYDRASTIS CANADENSIS L. USING RAPD ANALYSIS FEBRUARY 2009 KERRY J. KELLEY, B.A. MOUNT HOLYOKE COLLEGE M.A. UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Lyle Craker Hydrastis canadensis L. (goldenseal) is an endangered perennial wildflower species native to eastern North America. In this study, several populations of goldenseal, (both cultivated and wild type) were analyzed for genetic variability. The samples were collected from plant populations in North Carolina, Ohio, Pennsylvania and West Virginia and preserved using silica gel during collection. Random amplified polymorphic DNA (RAPD) analysis technique was used to generate DNA profiles from individual plants and to estimate genetic variability between groups (cultivated and wild type), among populations within groups and within populations using analysis of molecular variance (AMOVA) and a UPGMA clustering phenogram. Our results demonstrate that the bulk of genetic diversity may be within and among populations, but not between groups. This indicates the need for preservation and conservation efforts at the population level. The next step would be to study goldenseal populations more in depth for underlying causes of the genetic variability observed in this study. Further study of genetic variability with different molecular markers may be needed to clarify the level of diversity for the species at the group level. Increased knowledge of genetic variability and the identification of accessions of goldenseal would prove useful for reintroduction and cultivation strategies.

The following study was conducted to investigate the antibacterial and feed additive potential of medicinal plants. Ethanol extracts of different medicinal plants including Curcuma longa (Turmeric), Zingiber officinale (Ginger), Piper nigrum (Black Pepper), Cinnamomum cassia (Cinnamon), Thymus vulgaris (Thyme), Laurus nobilis (Bay leaf), and Syzgium aromaticum (Clove) were tested using the disc diffusion method for their antimicrobial activity against the common poultry pathogens E. coli, S. typhimurium, E. faecium, and E. faecalis. Cinnamon extract (CE), at 130 mg/disk, exhibited antibacterial activity against E. coli, S. typhimurium, and E. faecalis. Thyme extract (TE), at 30 mg/disk, exhibited antibacterial activity against E. coli, E. faecium, and E. faecalis while the remaining medicinal plants extracts showed no activity. The minimum inhibitory concentration (MIC) of the cinnamon and thyme ranged from 31.25 to 250 mg/ml by the dilution method. From this in vitro antibacterial study, cinnamon and thyme were selected for a 21-d feeding trial in broilers to study their influence on feed consumption, body weight gain, and feed conversion. There were 6 dietary treatments groups: 1) negative control (NC) containing no plant extracts or antibiotic, 2) positive control (PC) containing BMD (bacitracin) at 50g/ton of feed, 3) Diet 1 plus low level of cinnamon extract (LCE) at 290 gm/100 kg of feed, 4) Diet 1 plus high level of cinnamon extract (HCE) at 580 gm/ 100 kg of feed, 5) Diet 1 plus low level of thyme extract (LTE) at 290 gm/100kg of feed, and 6) Diet 6 plus high level of thyme extract (HTE) at 580 gm/100 kg of feed. No significant changes in body weight gain were observed with the cinnamon extracts compared to the NC or PC at 7, 14, or 21 d. The HTE reduced body weight gain compare to the NC and PC at 7, 14, and 21 d (P < 0.02). No difference in feed efficiency was observed with any of the treatments except LCE which reduced feed efficiency compared to other treatments. No difference in feed consumption was found among any of the treatments. These results suggest that cinnamon and thyme have antibacterial activity in vitro, and thyme has an activity that reduces body weight. Since cinnamon caused no significant change in body weight gain compared to positive or negative controls, it warrants further study as a substitute for antibiotics in the diet. / Master of Science

Orientador: Maria Aparecida Silva / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Química / Made available in DSpace on 2018-08-24T23:58:11Z (GMT). No. of bitstreams: 1 Silva_MateusGuimaraesda_M.pdf: 2502848 bytes, checksum: 5c28fb331431c609c6b31a9c93a01889 (MD5) Previous issue date: 2014 / Resumo: O guaco (Mikania laevigata Schultz Bip. Ex Baker) é uma das plantas medicinais mais conhecidas e difundidas para o tratamento de doenças respiratórias pela sua ação expectorante e broncodilatadora. A cumarina, presente na sua composição química, é um dos principais componentes responsáveis por tal atividade e utilizada como marcador químico no controle de qualidade dos produtos derivados de guaco. Os processos tradicionais de secagem utilizados para a conservação de folhas de guaco podem promover a degradação da cumarina, causando prejuízos para os fins fitoterápicos. Por isso, torna-se de grande importância o estudo de diferentes métodos de secagem das folhas de guaco de forma a minimizar a perda da cumarina. No presente trabalho, foi estudada a influência do etanol na secagem de folhas de guaco e no rendimento de cumarina e, também, avaliado a estabilidade da cumarina em amostras secas, durante o armazenamento em atmosfera acelerada e normal. Os experimentos de secagem foram realizados em uma estufa com circulação forçada de ar e em túnel de secagem com folhas inteiras e cortadas, respectivamente. Quando se utilizou etanol, as folhas enviadas ao processo de secagem foram mergulhadas em álcool etílico absoluto antes do início da secagem. Os resultados mostraram que as folhas submetidas ao processo de secagem usando etanol apresentaram os menores tempos de secagem e os maiores rendimentos de cumarina, tanto nos experimentos realizados no túnel quanto na estufa. Nos experimentos de secagem realizados com folhas cortadas, foi encontrada uma redução do tempo da ordem de 35% e nas folhas inteiras da ordem de 30%. Na análise do princípio ativo após os experimentos de secagem, foram encontrados os maiores rendimentos de cumarina nas folhas inteiras comparado às folhas cortadas. Nos experimentos de armazenamento, foi observada a degradação da cumarina ao longo do tempo, sendo mais pronunciada nas amostras secas sem etanol e em atmosfera acelerada / Abstract: The Mikania laevigata Schultz Bip. Ex Baker is a medicinal plant, popularly known as "guaco" in Brazil, it is widespread used in the treatment of respiratory diseases because of expectorant and bronchodilator effects. Coumarin is an active principle found in guaco, considered one of the main species responsible for this activity and used as a chemical marker for the quality control of products derived from guaco. The drying of guaco leaves is a process aimed at the preservation of substances, carried out to meet the needs of the herbal medicines industry which has no means to use fresh plants in the quantities required for industrial production. Conventional drying processes used for the preservation of guaco leaves may promote the degradation of coumarin, causing losses for phytotherapeutic purposes. Therefore, it is of great importance the study of different drying methods of guaco leaves to minimize the coumarin loss. In this work, it was studied the influence of ethanol on the drying kinetics and on the coumarin yield of guaco leaves and also evaluated the stability of coumarin of dried samples during storage under normal and accelerated atmosphere. The drying experiments were performed in a forced convection oven and tunnel drying with whole and cut leaves, respectively. The guaco leaves were dried with and without the surface previously treated with ethanol. The results showed that samples treated with ethanol presented faster water evaporation during drying experiments and higher yield of coumarin. In drying experiments carried out with cut leaves, the decrease in the drying time was approximately 35% and for whole leaves was approximately 30%. In the analysis of the active ingredient after drying experiments, the coumarin yield was higher in whole leaves than in cut leaves, around two times more. In the experiments storage, it was observed the coumarin degradation over time, being more pronounced in the samples dried without ethanol and under accelerated atmosphere / Mestrado / Engenharia Química / Mestre em Engenharia Química

Thesis (MScConsEcol (Conservation Ecology and Entomology))--Stellenbosch University, 2008. / Medicinal plants represent an important asset to the livelihoods of many people in developing countries. This is the case for South Africa where most of the rural and also urban communities rely on medicinal plants for their primary healthcare needs and income generation. Harvesting for domestic usage is not generally detrimental to the wild populations of medicinal plants. However, the shift from subsistence to commercial harvesting is posing unprecedented extinction threat to the wild populations of medicinal plants. The purpose of this investigation was to: (1) document the most traded/used species of medicinal plants in the Cape Peninsula, including parts used, sourcing regions, harvesting frequencies and seasons as well as the conservation status of these species; (2) to profile and investigate the rationales for the involvement of stakeholders in medicinal plants related-activities; and to (3) assess constraints and opportunities for sustainable management of medicinal plants in the Cape Peninsula. Triangulation techniques such as semi-structured questionnaires, formal and informal interactions with key informants from the Cape Peninsula and surroundings, personal observations and field visits were used to gather relevant data for this investigation. Accordingly, about 170 medicinal plant species were found to be actively traded or used in the study area. These species were mostly traded/used for their underground parts; shoot, barks and in many cases the whole plant is uprooted. The bulk of traded/used species were from the wild populations, harvested on monthly basis and the Western and Eastern Cape provinces acted as the main source regions. Some of the traded/used species are rare, vulnerable, endangered, critically endangered and are declining from the wild. Nonetheless, there are subtitutes for some of these medicinal plant species. Traders and collectors were mainly men in the Cape Peninsula. Cultural considerations, economic conditions and the burden imposed by the number of dependents were the factors influencing local communities to engage in medicinal plants related-activities. Despite the fact that the majority of the informants acknowledged the decline of medicinal plants from wild stocks, an overwhelming number of them expected an upsurge in the future demand for natural remedy due to its popularity among South Africans. Similarly, the majority of the respondents were aware of the conservation status of the plants that they were using, but this did not prevent them from trading/using some protected species. Encouragingly, an overwhelming number of the informants were willing to use cultivated species and cultivate some of the most used medicinal plant species if seeds and land were freely provided. It is noteworthy that these results were influenced by the gender, age, category and time of involvement in medicinal plants, ethnicity and residence status of the respondents as well as the source of supply of medicinal plants. It is recommended that species that have been identified of concern should be prevented from further commercial harvesting. Competent conservation organizations like CapeNature should focus on practical skills development of people who have expressed willingness to cultivate medicinal plants or are already doing so, especially in plant propagation and basic gardening techniques.

Thesis (PhD)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Plant secondary compounds have been exploited as medicines, flavourants, incense and are widely used for different culinary purposes. Efficacy of herbal remedies is determined by the phytochemical profile which is dependent on the species, locality and growth conditions. Salvia stenophylla (Burch. ex Benth.) is a local aromatic sage growing in almost all South African biomes, with the volatile compound accumulation varying depending on season, locality and genotype. Salvia stenophylla extracts have proven anti-inflammatory, anti-plasmodial, anti-oxidant and anti-anxiety properties due to the presence of essential oils, flavonoids and caffeic acid derived phytochemicals. As a result, S. stenophylla extracts are increasingly being used for the formulation of pharmaceutical and cosmeceutical products. However, these industries largely depend on the wild populations for raw materials, and increased commercialization of sage-based natural products and medicine exerts harvesting pressure. This might reduce the sustainability of herbal medicines since there is no formal cultivation of S. stenophylla in South Africa. Medicinal plant biotechnology provides options for propagation and manipulation of herbal plant to increase synthesis of secondary compounds. The aims of this study were to develop tissue culture propagation system so as to provide an alternative to wild-harvesting, to improve essential oil quality and accumulation in S. stenophylla by applying third generation biotechnological tools to alter terpene biosynthesis via heterologous gene expression aided by Agrobacterium-gene transfer and to charaterize the resultant metabolite profiles using thin layer chromatography, gas chromatography mass spectrometry and liquid chromatography mass spectrometry. Initially, seeds were germinated in-vitro on one-tenth Murashige and Skoog medium (MS) (1962). Seeds were decontaminated by washing them in 3.5% (w/v) hypochloride and were either scarified using 70% (v/v) sulphuric acid, placed on media with smoke solution or both. Both scarification and smoke supplementation induced germination, but seeds subjected to both treatments, as a combination, showed very poor germination. Seedlings produced were used to establish an efficient tissue culture system for S. stenophylla. Shoot tips, nodal and basal explants were placed on MS media with different plant growth regulator (PGR) combinations and concentrations. The best plantlet regeneration and shoot elongation were observed on plants on medium supplemented with 5.7 μM indole acetic acid (IAA) and 8.9 μM N-6-benzyladenine (BA), producing 4-6 shoots per explant with 6.67 cm mean length. Treatment with 4.5 μM 2,4 dichlorophenoxyacetic acid (2,4 D) did not produce shoots but had the highest biomass production, which was all in the form of callus. The resultant shoots from the micropropation system had similar metabolites as wild type plant and also they had some compounds that were not detected in their wild type counterparts. The effects of nitrogen, potassium, water stress, and phytohormones on metabolite accumulation were also studied in a S. stenophylla microplant system. Plant growth regulator free half strength MS medium encouraged the most prolific growth. Increasing potassium and nitrogen concentrations correlated to the accumulation of (-)-α-bisabolol whilst changes in other major compounds like ß-bisabolene, α-muurolene, α-patchoulene, and D-limonene were insignificant. To investigate the effects of water stress on metabolite accumulation and profile, water stress was induced using sorbitol and polyethyl glycol. Reduced water availability only negatively affected rooting in-vitro whilst the chemical profile was not affected. The study also focused on terpene production in S. stenophylla by over-expressing the heterologous geranyl diphosphate synthase gene (AgGPPS2) from Abies grandis, using Agrobacterium tumefaciens (EHA105). The AgGPPS2 gene was ligated to a pCAMBIA1301 vector which was cloned in EHA105 and this in turn was used to transform S. stenophylla. Resultant transgenic plantlets exhibited normal growth characteristics but showed variation in the metabolite accumulation. GC-MS analysis showed a 6% increase in (-)-α-bisabolol accumulation whilst 3-δ-carene, α-pinene and camphor were lowered. Solvent extracts were analysed using LC-MS and these had rosmarinic acid in greater abundance in transgenic than in the wild type plants. However, the chemical profiles of the wild type and transgenic plants showed some similarities suggesting AgGPPS2 expression only alters the abundance of some secondary compounds, whilst the overall integrity of the metabolome is maintained. Unlike most metabolite engineering studies elsewhere, this is the first attempt a local South African sage has been genetically engineered to enhance terpene biosynthesis. This is crucial in an economy that is increasing becoming plant based for pharmaceutical, industrial and food needs. Further studies are required to elucidate the efficacy of transgenic and in-vitro plant-derived extracts. / AFRIKAANSE OPSOMMING: Plant sekondêre produkte word gebruik as medisinale-, geur-en reukmiddels en word ook as algemene bestandeel gebruik in kookkuns. Die doeltreffendheid van kruie middels word bepaal deur die fitochemiese profiel. Hierdie profiel is afhanklik van die spesie, ligging en groei kondisies. Salvia stenophylla (Burch. ex Benth.) is ‘n plaaslike aromatiese salie wat voorkom in byna al die biome van Suid-Afrika , waar die versameling van vlugtige verbindings varieer op grond van die seisoen, ligging en genotipe. Salvia stenophylla ekstraksies is bewys om anti-inflamatories, antiplasmodiaal, anti-oksidant en anti-angs eienskappe te besit as gevolg van die noodsaaklike olies, flavonoïede en afgeleide kaffieksuur fitochemikalieë wat die plant besit. As gevolg word S. stenophylla ekstraksies meer en meer aangewend in die ontwikkeling van farmaseutiese en kosmetiese produkte. Hierdie industrieë benodig rou plant material vanuit die natuurlike populasies wat druk op die biome sit as gevolg van die verhoogde kommersialisering van salie-gebaseerde natuurlike produkte en medisyne. Gevolglik is die volhoubaarheid van medisyne afkomstig vanaf kruie soos salie in bedwang omdat geen formele kultivasie van S. stenophylla in Suid-Afrika al in plek gestel is nie. Medisinale plant biotegnologie bied moontlikhede vir kultiveering, voortplanting en manipuleering van kruie om sodoende die produksie van sekondêre verbindings in kruie te verhoog. Die doel van hierdie studie was om ‘n weefselkultuur voortplantingssisteem te ontwikkel om sodoende ‘n alternatief te bied vir wild oesting. Die studie het gefokus op die verbetering van noodsaaklike olie kwaliteit en akkumulasie in S. stenophylla deur die gebruik van derde generasie biotegnologiese gereedskap om terpeen biosintese te modifiseer. Hierdie is bereik deur weefselkultuur en heterologiese geen uitdrukking wat aangehelp is deur Agrobacterium geen oordrag. Die gevolglike metaboliet profiele was gekarakteriseer deur gebruik te maak van dun laag chromatogragie, gas chromatografie massa spektrometrie en vloeistof chromatografie massa spektrometrie. Aanvanklik is die sade ontkiem in-vitro op een-tiende Murashige en Skoog medium (MS) (1962). Dekontaminasie van die sade is gedoen deur die sade te was in 3.5% (w/v) hipochloried waarna die sade óf geskarifideer is deur te was met 70% (v/v) swawelsuur óf op medium geplaas is wat ‘n rook oplossing bevat óf albei. Beide skarifikasie en rook aanvulling het ontkieming aangehelp, maar sade wat aan albei behandelings blootgestel is, het baie swak ontkieming getoon. Saailinge was gebruik om ‘n effektiewe weefselkultuursisteem vir S. stenophylla te produseer. Stingel punte, nodale en basale eksplante was op MS medium geplaas met verskillende kombinasies en konsentrasies van verskeie plant groei reguleerders (PGR). Die beste plant regenerasie en stingel verlenging was geobserveer met plante op medium aangevul met 5.7 μM indool asynsuur (IAS) en 8.9 μM N-6-bensieladenien (BA), waar dit 4 tot 6 stingels per plant met ‘n gemiddelde lengte van 6.67 cm gegroei het. Behandeling met 2,4 Dichlorophenoksiasynsuur (2,4 D) het nie stingels produseer nie, maar het die hoogste biomassa produksie getoon in die vorm van kallus. Die gevolglike stingels van die mikrovoortplantingsisteem het soortgelyke metaboliete as die wilde tipe plante, maar verbindings wat nie in die wilde tipe plante voorkom nie was ook ontdek. Die effek van stikstof, kalium, water stres en fitohormone op metaboliet akkumulasie was ook bestudeer in S. stenophylla mikroplantsisteem. Plant groei reguleerder vrye half sterkte MS medium het die produktiefste groei aangemoedig. Verhoging van kalium en stikstof konsentrasies het gekorreleer met die versameling van (-)-α-bisabolol en terwyl veranderinge in ander belangrike verbindings soos ß-bisaboleen, α-muuroleen, α -patchouleen en D-limoneen onbeduidend was. Om die effek van water stres op metaboliet- akkumulasie en -profiel te ondersoek was water stres geinduseer deur sorbitol en poli-etielglikol te gebruik. Die verlaging van water beskikbaarheid het slegs ‘n negatiewe invloed op in-vitro wortelvorming gehad terwyl die chemiese profiel onveranderd gebly het. Die studie het ook gefokus op die terpeen produksie in S. stenophylla deur die ooruitdrukking van die heterologiese geranieldifosfaatsintase geen (AgGPPS2) vanaf Abies grandis met behulp van Agrobacterium tumefaciens (EHA105). Die AgGPPS2 geen was geligeer in ‘n pCAMVIA1301 vektor wat geklooneer was in EHA105 en dit op sy beurt was getransformeer in S. stenophylla. Die gevolglike transgeniese plante het normale groei ten toongestel, maar het variase in metaboliet akkumulasie getoon. GC-MS analises het gewys dat ‘n 6% toename in (-)-α-bisabolol versameling terwyl 3-δ-kareen, α-pineen en kamfor verlaag het. Opgelosde ekstraksies was geanaliseer deur VC-MS en het gewys dat roosmariensuur in groter hoeveelhede in transgeniese plante as wilde tipe plante akkumuleer. Nogtans het die chemiese profiele van die twee genotipes sommige ooreenkomste getoon, wat voorstel dat AgGPPS2 uitdrukking slegs sekere sekondêre verbindings menigte male vermeerder terwyl die algehele integriteit van die metaboloom gehoue bly. In teenstelling met die meederheid studies oor metaboliet ingenieuring wat elders plaasvind, is hierdie die eerste poging om ‘n plaalike Suid-Afrikaanse salie genieties te ingenieur met die oog om die terpeen biosintese te verbeter. Hiedie is krities in ‘n ekonomie wat voortdurend vorder om plant gebaseerde produkte te maak vir farmaseuties, industrieël en voedsel behoeftes. Verdere studies is nodig om helderheid te gee oor die effektiewiteit van transgeniese en in-vitro plant afkomstige ekstraksies.

Exploitation of botanical resources has resulted in significant decreases in the sizes of some plant populations, especially for species that have a high commercial value and are important to the lives and livelihoods of rural communities. Medicinal plant resources are used and traded commercially in both rural areas and urban centres, and over-exploitation has become a deterministic factor in the extinction risks to certain species. The main aim of the study was to design a risk assessment model to determine the impact of the medicinal plant trade on the Witwatersrand (centred around Johannesburg) on indigenous plant resources. The goal was to incorporate trade variables correlated with harvesting risks together with biological characteristics of the harvested species to predict which species are most threatened by the trade and are thus high on the list for conservation priority. The study required semi-quantitative surveys of the medicinal plants sold by traders in the Witwatersrand to be conducted. In 1994 and 2001, the plants sold in 50 muti shops and by 100 vendors in the Faraday Street market respectively were inventoried. Quantitative trade data were also captured, including volume, pricing structures and plant size (e.g. bark thickness and bulb diameter). A scientific sampling strategy was adhered to throughout the study to add statistical validity to the results. In a novel approach to analysing ethnobotanical data, the frequency of plant occurrences in the markets was analysed using measures (analysed by EstimateS) of species diversity traditionally used in ecology. The measures allowed for sampling strategies and sizes to be compared between data sets and for the number of species likely to be sold in the region to be estimated. Furthermore, data sets could be compared in terms of species richness, diversity, evenness and complimentarity. Another novel approach taken in the thesis was to estimate the number of individual plants harvested annually by gatherers, specifically the number of trees that are debarked and the number of whole bulbs that are removed. In order to estimate the number of trees debarked, a study was conducted to determine the relationship between bark thickness and stem diameter for six species. The results made it possible to estimate the condition of the resource in the wild from market records (i.e. bark thicknesses) and to see how the availability of larger trees has declined for species such as Warburgia salutaris between 1994 and 2001. Results for bulbs showed that there has been a significant decrease in the diameter of Eucomis autumnalis bulbs present in the markets in the same period, suggesting significant levels of resource depletion. The thesis explored the use of a multivariate methodology for assessing the extinction risks of species and assigning species harvested for the medicinal plant trade to various hierarchies of risk and conservation priority. Hierarchical and non-hierarchical cluster analysis (Ward’s and K-means respectively) methods were found to be effective in assigning species to clusters of similar risk and conservation priority. From a combined list of 392 ethnospecies recorded in the muti shops and Faraday market, a short-list of 119 higher risk species was identified using four to five trade variables. This list was further reduced to 87 species to ascertain conservation priorities based on the additional inclusion of seven biological variables in the assessment. From this list, approximately 31 species were identified as having higher conservation priority and would be candidates for further research, management and protection within the ambit of conservation and sustainable utilisation programmes. These species would further benefit from Orange Listing or having their IUCN Red List status re-evaluated. The methods developed in this study are recommended for other ethnobotanical studies. Furthermore, the risk assessment method could be applied to the assessment of species similarly traded in other medicinal plant markets or applied to the assessment of species under threat from other stressors at a regional, provincial and/or national level using the appropriate variables.

A calêndula é uma planta medicinal de grande importância para indústria cosmética e farmacêutica. Existem poucos estudos sobre as necessidades hídricas e nutricionais das plantas medicinais. O controle do potencial matricial de água no solo pode ser uma alternativa de manejo de irrigação que favoreça a qualidade e produção de metabólitos secundários. O objetivo do experimento foi avaliar a resposta fisiológica e bioquímica da Calendula officinalis L. submetida a vários níveis de potencial matricial de água no solo para manejo da irrigação em três diferentes fases de desenvolvimento. O experimento foi conduzido em ambiente protegido sem controle de temperatura e umidade na Escola Superior de Agricultura \"Luiz de Queiroz\", Piracicaba, São Paulo. O delineamento experimental foi em blocos ao acaso com 6 tratamentos e 4 repetições, totalizando 24 unidades experimentais. Foram utilizadas sementes de calêndula bonina sortida para produção de mudas. As mudas foram transplantadas para vasos de 8 L para condução do experimento. Os potenciais matriciais adotados foram: -10, -15, -30, -40, -50 e -60 kPa. Foram realizados três ensaios distintos definidos de acordo com a fase de desenvolvimento: ciclo completo, fase vegetativa e fase reprodutiva/florescimento. O controle do potencial matricial de água no solo foi realizado utilizando tensiômetros de punção previamente instalados na profundidade de 20 cm. As leituras foram realizadas uma vez por semana com auxílio de tensímetro, desde o transplantio até a senescência das plantas. As irrigações foram realizadas por sistema de irrigação por gotejamento com gotejadores autocompensantes com vazão média de 3,92 L h-1. Os parâmetros analisados foram: taxa de crescimento relativo, massa seca de parte aérea, comprimento de raiz, volume de raiz, massa seca de raiz, produção de capítulos florais, massa seca de capítulos florais, relação raiz/parte aérea, produtividade de água, teor e rendimento de flavonoides totais e concentração de prolina. Os dados obtidos foram submetidos ao teste F, ao teste de normalidade de Komogorov-Smirnov e ao teste de Scott- Knott para comparação de médias. Os parâmetros que apresentaram diferenças entre os tratamentos foram submetidos à regressão. Foi utilizado o pacote estatístico Assistat 7.7 Beta. A taxa de crescimento relativo, a massa seca de parte aérea, o volume e massa seca de raiz, e a produção de capítulos florais apresentaram diferenças significativas apenas para o ensaio com estresse hídrico durante todo ciclo de desenvolvimento da calêndula. Para essas variáveis a diminuição da disponibilidade de água no solo apresentou resultados negativos para a planta. A massa seca de capítulos florais foi superior para o potencial matricial de -10 kPa durante a fase reprodutiva e ciclo completo. A produtividade de água foi superior para os potenciais matriciais de -30 e -40 kPa no ciclo completo e fase reprodutiva. O rendimento de flavonoides e a concentração de prolina apresentaram resultados inversamente proporcionais. A redução do rendimento de flavonoides com o decréscimo da disponibilidade de água no solo favoreceu o aumento da prolina. / Calendula is a medicinal plant of great importance in cosmetic and pharmaceutical industry. There are few studies on hydric and nutritional requirement of medicinal plants. The control of water with soil matric potential can be an alternative to the irrigation management that improve the quality and production of secondary metabolites. The objective of the experiment was to evaluate the physiological and biochemical response of Calendula officinalis L. subjected to various degree of water stress soil water for irrigation management in three different stages of development. The experiment was conducted in a greenhouse without temperature and humidity control in the Escola Superior de Agricultura \"Luiz de Queiroz\", Piracicaba, São Paulo. The experimental design was a randomized block design with 6 treatments and 4 repetitions, totaling 24 experimental units. The calendula bonina sortida seed were used to produce seedlings. The seedlings were transplanted to 8 L vessels to conduct the experiment. The potential matrices adopted were: -10, -15, -30, -40, -50 and -60 kPa. Three different tests were performed according to defined stages of development: full cycle, vegetative phase and reproductive/flowering stage. The control of the matric potential of soil water was performed using punch tensiometers previously installed at a depth of 20 cm. The measurements were performed with a tensiometer once a week, beginning the transplanting until senescence of plants. The irrigation was drip irrigation system with drip with flow of 4 L h-1. The parameters analyzed were: relative growth rate, dry weight of shoot, root length, root volume, root dry weight, production of floral chapters, dry mass of flower heads, root/shoot, water productivity, content and yield of total flavonoids and concentration of proline. The data were submitted to the test F, the normality test Komogorov -Smirnov and the Scott -Knott test for comparison of means. The parameters that show differences between treatments were submitted to regression. We used the statistical package Assistat 7.7 Beta . The relative growth rate, shoot dry weight, volume and root dry weight, and the production of floral chapter showed significant differences only for test water stress throughout the marigold development cycle. For these variables, decreased water availability in the soil was negative for the plant. The dry mass of floral chapter was superior to the matric potential of - 10 kPa during the reproductive phase and full cycle. The productivity of water was higher for the matric potentials of -30 and -40 kPa in the complete cycle and reproductive phase. The yield of flavonoids and proline concentration showed inversely proportional results. The reduction in the yield of flavonoids with decreased water availability occurred an increase of proline.

A Uncaria tomentosa (U. tomentosa), popularmente conhecida como \"Unha-de-gato\", é uma planta medicinal nativa das Américas, mundialmente empregada devido às suas atividades anti-inflamatórias e imunomodulatórias. O consumo desta planta ocorre não apenas na forma in natura, mas principalmente como fitoterápico, sendo muitas vezes utilizada de forma indiscriminada pela população. Apesar de vários estudos revelarem as propriedades terapêuticas da U. tomentosa, poucos são os trabalhos que empregam protocolos estabelecidos por agências regulamentadoras internacionais, para a avaliação dos possíveis efeitos tóxicos e imunotóxicos deste fitoterápico. Assim, o propósito do presente estudo foi verificar se a administração de um extrato seco de U. tomentosa, comercialmente disponível no mercado, poderia ocasionar efeitos tóxicos e imunotóxicos em ratos após 90 dias de tratamento. Para isso, 40 ratos Wistars machos foram tratados oralmente com as doses de 15, 75 ou 150 mg/kg de extrato seco de U. tomentosa comercialmente disponível no mercado, contendo teores de alcaloides de acordo com aqueles valores preconizados em literatura. No final do período experimental, os animais foram submetidos à eutanásia para realização de avaliações bioquímicas, hematológicas, histopatológicas, análise de órgãos linfoides e não-linfoides, avaliação das respostas imunes inata, inflamatória e humoral, bem como teste para determinação de reação de hipersensibilidade do tipo IV. Os resultados revelaram aumento nos níveis de ALT dos animais tratados com a dose de 75 mg/kg, e redução nos índices glicêmicos de ratos tratados com 75 e 150 mg/kg de U. tomentosa. Entretanto, somente os ratos tratados com a maior dose exibiram discreta vacuolização centro-lobular hepática; assim, somente os dados de ALT não são sugestivos de efeitos hepáticos adversos da U. tomentosa após um longo período de tratamento. A redução nos índices sanguíneos de glicose dos ratos, após tratamento com a U. tomentosa, podem representar importante risco para seres humanos diabéticos, susceptíveis ao desenvolvimento de hipoglicemia e que fazem uso da U. tomentosa para outros propósitos. Em conclusão, estes estudos demonstraram que, apesar de a U. tomentosa não promover efeitos tóxicos e imunotóxicos, o uso prolongado da mesma, a altas doses, pode promover redução dos índices glicêmicos. / Uncaria tomentosa (U. tomentosa), commonly known as \"Cat\'s claw\", is a native medicinal plant from America, it is employed worldwide for its anti-inflammatory and immunomodulatory activities. The consumption of this plant occurs not only in natura, but mainly as a phytotherapic, used indiscriminately by the population. Although many researchers revealed the therapeutic properties of U. tomentosa, few studies employing established protocols by international regulatory agencies for the evaluation of the possible toxic and immunotoxic effects of this herbal medicine. Thus, the purpose of the present study was to verify if the dry extract of U. tomentosa could promote toxic and/or immunotoxic effects in rats following 90 days of treatment. For this, forty male rats were orally treated with 15, 75 or 150mg/kg of dry extract of U. tomentosa, commercially available, containing levels of alkaloids according to those values recommended in the literature. At the end of experimental period, the rats were killed for the evaluation of the biochemistry, haematology, histopathology, status of the lymphoid and non-lymphoid organs, evaluation of innate, inflammatory and humoral immune responses, as well as a test to determine the delayed type hypersensitivity. The results revealed an increase in the levels of ALT in the animals treated with 75mg/kg and a reduction in the glycaemic levels of rats treated with 75 and 150mg/kg of U. tomentosa. However, only rats treated with the higher dose showed a slight centrilobular hepatic vacuolation; thus, ALT data alone are not suggestive of a hepatic adverse effect of U. tomentosa following long-term treatment. The reduction in blood glucose levels of the rats, could represent an important risk for diabetic humans, who are susceptible to the development of hypoglycaemia and who might use U. tomentosa for purposes other than anti-diabetes. In conclusion, these studies demonstrated that, while U. tomentosa has no immunotoxic effect, long-term U. tomentosa treatment at high doses can promote reduction in glycemic levels.

Submitted by Michele Mologni (mologni@unoeste.br) on 2019-01-24T15:51:50Z No. of bitstreams: 1 Luís Gustavo Torquato Feba.pdf: 2538384 bytes, checksum: e99480491242ecf61572f38231c04aa7 (MD5) / Made available in DSpace on 2019-01-24T15:51:50Z (GMT). No. of bitstreams: 1 Luís Gustavo Torquato Feba.pdf: 2538384 bytes, checksum: e99480491242ecf61572f38231c04aa7 (MD5) Previous issue date: 2018-11-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The objective of this research was to evaluate the production of passion fruit (Passiflora incarnata L.) in the edaphoclimatic conditions of Presidente Prudente-SP, as well as its development in sandy soil under the effect of organic fertilization. A randomized block design with six treatments and four replicates was used. Were tested 3 kinds of fertilizers (fertilizer without control, cattle manure - 7.6 t ha-1 and chicken manure - 4.2 t ha-1) and two cutting times (at 90 and 150 days after planting) . At each cutting season the dry biomass production (aerial part) was evaluated and total phenol content, total flavonoids and antioxidant activity in the ethanolic extract of the plants were analyzed. Soil chemical analyzes were performed for comparison with the initial condition and foliar analyzes for the macronutrient content for each cutting season. Dry matter production was not affected by the factors "fertilization" and "cutting season". The content of total phenolic compounds in leaves was higher in plants fertilized with cattle and chicken manure, which also resulted in higher antioxidant activity. Factors "fertilization" and "time" cutting exerted isolated effect on soil chemical properties at the end of the experiment, and the poultry manure provided increases in K, Mg and the sum of the ground bases and the second cut were observed lower contents of organic matter, S and K compared to the first cut. The average yield obtained in the treatments (1 t.ha-1) is within the expected for the crop. the wild passion fruit cultivation The conclusion that can be given to the state western region of São Paulo and the organic fertilizer via use of cattle and chicken waste has resulted in higher quality of the raw material due to the higher phenolic concentration in the leaves and higher antioxidant activity. It is recommended that the quantities of manure applied be greater than those recommended to result in biomass gains, depending on the sandy soil condition of the site, in which the decomposition of manures is faster. / O objetivo deste trabalho de pesquisa foi avaliar a produção do maracujá silvestre (Passiflora incarnata L.) nas condições edafoclimáticas de Presidente Prudente- SP, bem como o seu desenvolvimento em solo arenoso sob o efeito da adubação orgânica. Utilizou-se o delineamento de blocos ao acaso, com seis tratamentos e quatro repetições. Foram testados 3 tipos de adubação (controle sem adubação, esterco de gado – 7,6 toneladas ha-1 e esterco de galinha – 4,2 toneladas ha-1) e 2 épocas de corte (aos 90 e 150 dias após o plantio). Em cada época de corte foi avaliada a produção de biomassa seca (parte aérea) e foram realizadas análises quanto ao teor de fenóis totais, flavonoides totais e atividade antioxidante no extrato etanoico das plantas. Realizaram-se também análises químicas do solo, para comparação com a condição inicial e análises foliares quanto ao teor de macronutrientes para cada época de corte. A produção de massa seca não foi afetada pelos fatores “adubação” e “época de corte”. O teor de compostos fenólicos totais nas folhas foi maior nas plantas adubadas com estercos de gado e galinha, o que resultou também em maior atividade antioxidante. Os fatores “adubação” e “época” de corte exerceram efeito isolado sobre os atributos químicos do solo ao final do experimento, sendo que o esterco avícola proporcionou aumentos nos teores de K, Mg e na soma de bases do solo e no segundo corte foram observados menores teores de matéria orgânica, S e K em comparação ao primeiro corte. A média de produtividade obtida nos tratamentos (1 t.ha-1) encontra-se dentro do esperado para a cultura. Concluiu-se que o cultivo de maracujá silvestre pode ser indicado para a região Oeste do Estado de São Paulo e que a adubação orgânica via utilização de resíduos avícola e bovino resultou em maior qualidade da matéria prima, devido à maior concentração fenólica nas folhas e maior atividade antioxidante. Recomenda-se que as quantidades de esterco aplicadas sejam maiores que as recomendadas para resultar em ganhos de biomassa, em função da condição de solo arenoso do local, no qual a decomposição dos estercos é mais rápida.

Made available in DSpace on 2016-08-10T10:54:48Z (GMT). No. of bitstreams: 1 Dwight Assis Chaves.pdf: 1963972 bytes, checksum: a15ea173ac93c1534367c22dfc6105bd (MD5) Previous issue date: 2015-02-05 / The barbatimão (Stryphnodendron adstringens) is a medicinal plant, rich in tannins, found in the cerrado, which has anti-inflammatory, analgesic activity and a protective activity of gastric mucosa. In this work we evaluated the angiogenic activity and result from antiangiogenic, genotoxic and non-genotoxic of the barbatimão bark aqueous solution. Data from recent studies of the angiogênise using the CAM of hens egg to the barbatimão were not found in the literature. The barbatimão shells for the preparation of ASB (aqueous solution of barbatimão) were acquired in the city of Goiânia. Were used 30 grams of crushed shells to one litre of water. Angiogenic activities and result from antiangiogenic were evaluated by the experimental method of the corioalantoide membrane (CAM) and as positive control was used the Regederm product, which features angiogenic activity known. Genotoxic activity was evaluated from the barbatimão from aqueous solution using experimental model the micronucleus test in bone marrow of mice, and non-genotoxic activity was evaluated by the simultaneous treatment of the ASB and the compound known to be genotoxic Mitomycin C (MMC). The presented ASB tests, was considered genotoxic, angiogenic and features not non-genotoxic on concentration tested compared with the controls. The ASB presented a high vascularization in MCA (50.4) compared with the positive control (52.9). The dose of the barbatimãos peel solution (30 mg.ml -1) analyzed presented difference (p<0,05) at the EPCMN frequency relative to the negative control group.Comparing the frequency of dose EPCMN SABMMC concurrent treatment with the positive control, there was no significant difference (p>0,05). The results were promising, cooperating with future research for developing new medicines. It is noticed that the genus presents promising active principles for the development of new herbal medicines, that becomes necessary more studies and research for consumer safety. / O barbatimão (Stryphnodendron adstringens) é uma planta medicinal, rica em taninos, encontrada no cerrado,a qual possui atividade anti-inflamatória, analgésica e uma atividade protetora da mucosa gástrica. Neste trabalho foram avaliadas as atividades angiogênica e antiangiogênica, genotóxica e antigenotóxica da solução aquosa da casca do Barbatimão.Dados dos estudos recentes da angiogênise utilizando a MCA de ovo de galinha para o barbatimão não foram encontrados na literatura.As cascas de barbatimão para o preparo da SAB(Solução aquosa de barbatimão) foram adquiridas na cidade de Goiânia. Foram utilizadas 30 gramas de cascas trituradas para um litro de água. As atividades angiogênica e antiangiogênica foram avaliadas pelo método experimental da membrana corioalantoide (MCA) e como controle positivo foi utilizado o produto Regederm, que apresenta atividade angiogênica conhecida. A atividade genotóxica foi avaliada a partir da solução aquosa do barbatimão utilizando como modelo experimental o teste do micronúcleo em medula óssea de camundongos, sendo que a atividade antigenotóxica foi avaliada pelo tratamento simultâneo da SAB e do composto sabidamente genotóxico Mitomicina C (MMC). A SAB pelos testes apresentados foi considerada angiogênica, genotóxica e não possui características antigenotóxicas na concentração testada comparada com os controles. A SAB apresentou uma elevada vascularização na MCA (50.4%) quando comparada com o controle positivo(52,9%). A dose da solução da casca de barbatimão (30 mg.ml-1) analisada apresentou diferença (p<0,05) na frequência de EPCMN em relação ao grupo controle negativo.Comparando a frequência de EPCMN da dose do tratamento de SAB+MMC simultâneo com o controle positivo, não houve diferença significativa (p>0,05). Os resultados foram promissores, cooperando com pesquisas futuras para desenvolvimento de novos medicamentos.Percebe-se que o barbatimão apresenta princípios ativos promissores para o desenvolvimento de novos medicamentos fitoterápicos, para isso tornam-se necessários mais estudos e pesquisas para segurança do consumidor final.