Através de Testes de Diluição em Sistema de Tubos Múltiplos determinou-se a Intensidade de Atividade de Inibição Bacteriana (IINIB/bacteriostasia) e a Intensidade de Atividade de Inativação Bacteriana (IINAB/bactericidia) de soluções contendo extratos hidroetanólicos e hídricos (decocto e infuso) de três acessos de Bixa orellana L. (urucum) a saber: Arroio do Meio/RS, Eldorado do Sul/RS e Maringá/PR, sobre inóculos padronizados de Salmonella Enteritidis (ATCC 11076), Escherichia coli (ATCC 11229), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 19433) e Listeria monocytogenes (ATCC 19114). Determinou-se, paralelamente, o teor de bixina presente nas sementes. Os extratos hídricos apresentaram baixa atividade de inibição e/ou inativação sobre os inóculos bacterianos, enquanto que a forma de extração hidroetanólica apresentou atividade antibacteriana seletiva e significativamente mais intensa (inibição/inativação) entre as cinco bactérias testadas. Independente da forma de extração, as bactérias Enterococcus faecalis e Listeria monocytogenes foram as mais sensíveis à atividade antibacteriana, enquanto que Escherichia coli apresentou a menor sensibilidade. Houve diferença significativa entre os teores de bixina dos três acessos, e, consequentemente, a atividade antibacteriana determinada mostrou-se diretamente proporcional a estes teores. / Through of Dilution Tests in Multiple Tubes System it was determined the intensity of bacterial inhibition activity (IINIB/bacteriostasy) and the intensity of bacterial inactivation activity (IINAB/bactericidie) from solutions containing hidroetanolic and hydric (decoction and infusion) extracts of tree accesses from Bixa orellana L. (annatto) at know: Arroio do Meio/RS, Eldorado do Sul/RS and Maringá/PR, on standardized inocula of Salmonella Enteritidis (ATCC 11076), Escherichia coli (ATCC 11229), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC 19433) and Listeria monocytogenes (ATCC 19114). It was determined, in parallel, the content of bixin present in the three different accesses of the seeds. The forms of hydric extraction showed low inhibition and/or inactivation activity of the bacterial inocula, and the hidroetanolic extract form showed selective antibacterial activity and significantly pronounced inhibition/inactivation against the five bacteria tested. Independent of the extraction forms, Enterococcus faecalis and Listeria monocytogenes were the more sensitive agents to the antibacterial activity. Escherichia coli had the lowest sensitivity to all forms of extraction. The bixin contents were significantly different between the accesses and, consequently, the antibacterial activity was directly proportional to this contents.

Introduction : La résistance aux antibiotiques est une menace de santé, il est donc urgent de développer de nouveaux antibactériens. CIN-102, est un nouvel antibactérien développé par une industrie pharmaceutique. Il possède un large-spectre d’action et aucune résistance n’a été développée jusqu’à présent. Parmi les possibles applications thérapeutiques du CIN-102, notre recherche s’est focalisée sur les Maladies Inflammatoires Chroniques de l’Intestin (MICIs). Plusieurs facteurs contribuent à l’étiologie des MICIs. Les bactéries intestinales jouent un rôle important dans ces maladies et une augmentation de la charge bactérienne est observée pendant l’inflammation. Les objectifs de ce travail ont été : la caractérisation de l’activité antibactérienne du CIN-102, l’analyse de l’activité antibactérienne des agents anti-inflammatoires et antibiotiques utilisés en cas de MICI et la fabrication des formulations à ciblage colique pour le CIN-102. Le but est de diminuer la charge bactérienne colique par moyen du CIN-102 et améliorer, de cette façon, l’état de l’inflammation.Méthodes: La Concentration Minimale Inhibitrice (CMI), l’Effet Post-Antibiotique (EPA) et le temps de réduction logarithmique du CIN-102 ont été déterminés pour des bactéries aérobies et anaérobies. Les interactions entre le CIN-102 et des antibiotiques sur le marché ont été évaluées. La CMI de l’acide 5-aminosalicylique (5-ASA), GED-0507-34 et antibiotiques ont été déterminées pour des souches anaérobiques. Par rapport aux formulations à libération prolongée : des mini-granules contenant le CIN-102 ont été fabriqués par extrusion-sphéronisation puis pelliculés avec des mélanges de polymères insolubles et polysaccharides. Parallèlement, des mini-comprimés de CIN-102 ont été fabriqués par compression directe. La libération du CIN-102 in vitro, a été mesurée dans des milieux simulant l’estomac et l’intestin grêle. L’efficacité des systèmes à libération prolongée a été évaluée dans un modèle de colite chez la souris. Des prélèvements de selles et tissus coliques ont été soumis à des études bactériologiques. L’expression des cytokines a été mesurée à partir des tissus coliques.Résultats et discussion : Le large-spectre d’action du CIN-102 a été confirmé. Toutes les souches ont été inhibées par le CIN-102. CIN-102 présente un EPA et un temps de réduction logarithmique court. Il présente des interactions synergiques avec plusieurs antibiotiques, notamment la colistine et les aminoglycosides, en les rendant actifs contre des bactéries multirésistantes. Ces résultats in vitro doivent être poursuivis par des études chez l’animal. Des agents anti-inflammatoires utilisés contre les MICIs ne possèdent pas d’activité antibactérienne. Par ailleurs, les antibiotiques testés n’ont pas un large-spectre d’action contre des bactéries anaérobies généralement retrouvées dans l’intestin. Cela confirme le besoin d’un antibiotique à large spectre capable de réduire des charges bactériennes en cas d’inflammation. Dans ce but, des formulations capables de délivrer CIN-102 au niveau du colon ont été étudiées. La libération du CIN-102 des mini-granules pelliculés et mini-comprimés a été réduite dans des milieux simulant l’estomac et l’intestin grêle. Des souris atteintes de colite et traitées avec les formulations du CIN-102 ont eu une diminution des diarrhées et du sang dans les selles. Les concentrations d’entérobactéries adhérentes à la muqueuse colique et dans les selles ont été significativement réduites chez les souris traitées avec le CIN-102. Ces résultats montrent que ces formulations peuvent délivrer CIN-102 dans le tractus gastro-intestinal inferieur, et que la diminution d’entérobactéries semble réduire les symptômes de la colite.Conclusion : CIN-102 est un nouvel antibactérien a large-spectre et des formulations à libération prolongée peuvent délivrer cet agent dans le colon, diminuant la charge d’entérobactéries qui pourrait influencer l’état de l’inflammation. / Introduction: Antibiotic resistance is a major threat to public health and new antimicrobials are urgently needed. CIN-102, a new antibacterial agent which resembles cinnamon essential oils composition, was developed by a pharmaceutical company. CIN-102 had a broad-spectrum of action and resistance was not developed until now. Between all the possible therapeutic applications for CIN-102, a future utilization against Inflammatory Bowel Diseases (IBD) is aimed. IBD are chronic pathologies with a multifactorial etiology. In this context, enteric bacteria are well-known to have an important role, and higher bacterial concentrations are found in the intestine under inflammatory conditions. The objectives of this work were: to characterize the bacteriological activity of CIN-102, to analyze the bacteriological activity of anti-inflammatory agents and antibiotics used in IBD and to fabricate multiparticulate CIN-102 pharmaceutical forms for colonic targeted drug release. The idea is to use CIN-102 to reduce colonic bacterial loads and improve the state of intestinal inflammation. Methodology: The Minimal Inhibitory Concentration (MIC), the Post-Antibiotic Effect (PAE) and the logarithmic reduction time of CIN-102 were determined against several aerobic and anaerobic bacterial isolates. The interactions between CIN-102 and commercialized antibiotics were evaluated. The MIC of 5-aminosalicilyc acid, GED-0507-34 and antibiotics were determined for anaerobic bacterial isolates. Concerning sustained released formulations: CIN-102 pellet cores were fabricated by extrusion-spheronization and subsequently coated with blends of insoluble polymers and natural biodegradable polysaccharides. CIN-102 mini-tablets were fabricated by direct compression. In vitro drug released was measured in simulated gastric and intestinal fluid. The efficacy of best sustained release formulations was assessed in a murine model of colitis. Samples of luminal contents and sections of the colon were taken to perform a bacteriological analysis. Expression of cytokines was analyzed from colonic tissues.Results and discussion: The broad-spectrum activity of CIN-102 was confirmed. All aerobic and anaerobic strains were susceptible to CIN-102. Furthermore, CIN-102 had an important PAE and exerted a fast logarithmic reduction of bacterial inoculum. It interacts synergistically with several antibiotics, mostly with colistin and aminoglycosides, restoring the antibiotic activity against multi-resistant bacteria. The promising in vitro activity of CIN-102 has to be further confirmed by animal studies. Anti-inflammatory agents used against IBD were not provided of antibacterial activity and neither of the antibiotics tested possessed a broad-spectrum of action against anaerobic isolates commonly found in the intestine. These results confirm the need of a broad-spectrum antibiotic capable of reduced increased bacterial loads during inflammation. Following this aim, oral dosage forms able to deliver CIN-102 into the colon were studied. Concerning the sustained release forms, in vitro CIN-102 release from coated-pellets and mini-tablets was reduced in simulated gastric and intestinal fluids. Colitic mice treated with CIN-102 controlled release formulations had less diarrhea and bloody stools. Furthermore, the concentrations of enterobacteria in colonic tissue and stool were significantly reduced in CIN-102 treated mice. These results show that sustained release formulations can effectively deliver CIN-102 in the lower part of the gastrointestinal tract, where the reduction of enterobacteria seems to ameliorate the course of colitis.Conclusion: CIN-102 is novel broad-spectrum antibacterial and sustained release formulations can effectively deliver this agent into the colon, reducing bacterial loads which might influence the state of intestinal inflammation.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-06-26T17:25:10Z No. of bitstreams: 1 LediaWanderley.pdf: 1766339 bytes, checksum: 1ff559d6faf9483c7037afc6b631b271 (MD5) / Made available in DSpace on 2017-06-26T17:25:10Z (GMT). No. of bitstreams: 1 LediaWanderley.pdf: 1766339 bytes, checksum: 1ff559d6faf9483c7037afc6b631b271 (MD5) Previous issue date: 2016-12-21 / Fundação de Amparo à Pesquisa e ao Desenvolvimento Científico e Tecnológico do Maranhão (FAPEMA) / Helminthes and bacterial infections are a major challenge for livestock production because they cause loss of productivity in herds. To reduce losses, producers control parasites and bacteria mainly using synthetic chemicals, but the resistance to the active principles of these products calls for the continuous search for biologically active molecules. This research was conducted with the objective of obtaining protein fractions and protease from the latex of Ficus benjamina, in addition to evaluating the potential use of these protein samples in the control of the parasite nematode of small ruminants, Haemonchus contortus, as well as on the pathogenic bacteria Salmonella enterica, Staphylococcus aureus and Enterobacter aerogenes. Latex was obtained from an incision made at the apex of the plant limb and collected in sodium phosphate buffer (75 mM, pH 7.0). The latex was centrifuged (12,000 x g at 4 ° C for 15 min), dialysed against water (14 kDa cut-off), again centrifuged under the same conditions as before and then referred to as EPL (Latex Protein Extract). The EPL was fractionated with ammonium sulphate at 30-60% and 60-90% saturation to obtain the fractions denominated F30-60 and F60-90, respectively. F60-90 was fractionated by ion exchange chromatography on CM-Cellulose matrix and the fraction with the highest proteolytic activity obtained was denominated FbP (Ficus benjamina Protease). With these fractions, protein analysis, proteolytic activity and FbP identification were carried out. In addition, inhibition of larval development of the nematode H. contortus and inhibition of the growth of S. enterica, S. aureus and E. aerogenes in a liquid medium were carried out. The total protein levels obtained were, respectively, 121.5, 21.2, 75.7 mg for EPL, F30-60 and F60-90. The proteolytic activity of the protein samples was 43,471, 13,245 and 57,857 UA/mL/mgP for EPL, F30-60 and F60-90, respectively. The FbP presented proteolytic activity of 36,393.8 AU/mL/mgP. Gel analysis showed FbP with a protein band with mass between 20 and 30 kDa and by mass spectrometry the FbP was identified as cysteine protease, with a molecular weight of 23.97 kDa. FbP maintained stable proteolytic activity in the pH range between 6 and 10 and over a wide temperature range, with optimum activity at 60 °C. In tests for inhibition of larval development, EPL, F30-60, F60-90 and FbP presented IC50 of 0.22, 1.38, 0.42 and 0.26 mg/mL, respectively. It was verified that EPL, F30-60 and F60-90 did not exert an inhibitory action on the growth of bacteria, but FbP inhibited the growth of S. aureus and S. enterica with minimum inhibitory concentration (MIC) of 3.12 μg/mL. It is concluded that FbP obtained from latex of F. benjamina has promising biological properties for the development of products that can be used in the control of the gastrointestinal nematode, H. contortus, and the pathogenic bacteria S. aureus and S. enterica. / Helmintoses e infecções bacterianas são um grande desafio para a produção pecuária porque provocam perda de produtividade nos rebanhos. Para reduzir prejuízos, os produtores realizam o controle de parasitos e bactérias principalmente utilizando produtos químicos sintéticos, mas a resistência aos princípios ativos desses produtos insta a busca contínua por moléculas biologicamente ativas. Essa pesquisa foi conduzida com o objetivo de obter frações proteicas e protease a partir do látex de Ficus benjamina, além de avaliar uso potencial dessas amostras proteicas no controle do nematóide parasito de pequenos ruminantes, Haemonchus contortus, bem como sobre as bactérias patogênicas Salmonella enterica, Staphylococcus aureus e Enterobacter aerogenes. O látex foi obtido a partir de uma incisão feita no ápice do galho das plantas e coletado em tampão fosfato de sódio (75 mM, pH 7,0). O látex foi centrifugado (12.000 x g a 4 °C por 15 min), dialisado contra água (cut-off 14 kDa), novamente centrifugado nas mesmas condições anteriores e, então, denominado EPL (Extrato Proteico do Látex). O EPL foi fracionado com sulfato de amônio a 30-60% e 60-90% de saturação para a obtenção das frações denominadas F30-60 e F60-90, respectivamente. A F60-90 foi fracionada por cromatografia de troca iônica em matriz de CM-Cellulose e a fração com maior atividade proteolítica obtida foi denominada FbP (Ficus benjamina Protease). De posse dessas frações, foram conduzidos ensaios de análise de proteínas, atividade proteolítica e identificação da FbP. Adicionalmente foram realizados ensaios de inibição do desenvolvimento larvar do nematódeo H. contortus e inibição do crescimento das bactérias S. enterica, S. aureus e E. aerogenes em meio líquido. Os teores de proteína totais obtidos foram, respectivamente, 121,6, 21,2, 75,7 mg para EPL, F30-60 e F60-90. A atividade proteolítica das amostras proteicas foi de 43.471, 13.245 e 57.857 UA/mL/mgP para EPL, F30-60 e F60-90, respectivamente. A FbP apresentou atividade proteolítica de 36.393,8 UA/mL/mgP. A análise em gel mostrou a FbP com uma banda proteica com massa entre 20 e 30 kDa e por espectrometria de massa identificou-se a FbP como protease cisteínica, com massa molecular de 23,97 kDa. FbP manteve atividade proteolítica estável na faixa de pH entre 6 e 10 e em uma ampla faixa de temperatura, com atividade ótima à 60 °C. Em testes de inibição do desenvolvimento larvar, EPL, F30-60, F60-90 e a FbP apresentaram IC50 de 0,22, 1,38, 0,42 e 0,26 mg/mL, respectivamente. Foi verificado que EPL, F30-60 e F60-90 não exerceram ação inibitória sobre o crescimento das bactérias, mas FbP inibiu o crescimento de S. aureus e S. enterica com mínima concentração inibitória (MIC) de 3,12 μg/mL. Conclui-se que a FbP obtida a partir do látex de F. benjamina possui propriedades biológicas promissoras para o desenvolvimento de produtos que possam ser utilizados no controle do nematóide gastrointestinal, H. contortus, e das bactérias patogênicas S. aureus e S. enterica.

Antimicrobials are unique drugs in that they target "infectious" or "transferable" diseases. There is considerable evidence linking increasing antimicrobial use withincreasing resistance. Resistant bacteria do not know the boundaries, either between countries or within a society between hospital and primary care. Inappropriate prescribing of antimicrobials in hospitals therefore has consequences for whole communities and problems may spread both nationally and internationally. The gathering of reliable measurements of antibiotic use in hospitals employing standardised methods is essential to building an evidence base and highlighting inconsistencies at national and international levels. In this study, after data processing, validating and record linkage, a method forelectronic conversion of drug supply data to the ATC/DDD classification and forlongitudinal analysis was established for Tayside and then for a set of Europeanhospitals. Time series analysis and interrupted time series analysis were described and used for longitudinal surveillance and interventional study of antimicrobial use. This thesis explores issues concerning the evolution and management of hospital antimicrobial use using a wide range of methods. A series of drug utilisation research studies were implemented as the basis of research methods that, in combination of previously described methods, provided novel studies. No single measure can currently capture all of the aspects of hospital antibiotic use. However, a combination of detailed, point prevalence data from individual patients with longitudinal analysis of total consumption can provide meaningful data for comparison between hospitals and for analysis of the relationship between use and outcome. Additionally, there is a need to apply standard processes and novel methods to produce more meaningful surveillances. Longitudinal and point prevalence surveillances together with an explanation ofvariations in hospital characteristics are used to produce a set of coherent measurements of hospital antimicrobial use. Administrative data for longitudinal surveys requires continuous quality control.Whereas drug utilisation researchers and clinicians should target a set of indicators for interventional studies, large studies at national or international level need central data processing by country to identify targets for evaluation and for interventional studies. Support from experts in other fields is needed to address any shortcomings that may be experienced during continuous antibiotic drug utilisation monitoring at national and international levels.

The bacterium Rhodococcus is a potential source for novel antimicrobial metabolites. Recently, the Rhodococcus strain MTM3W5.2 was isolated from a soil sample collected from Morristown, East Tennessee and was found to produce an inhibitor molecule that is active against similar Rhodococcus species. The aim of this research is to extract, purify, and characterize the active compound. The compound was obtained from both agar and broth cultures of strain MTM3W5.2 and purified by primary fractionation of crude extract on a Sephadex LH-20 column, followed by semi-preparative reversed phase column chromatography. Final purification was achieved using multiple rounds of an analytical C18 HPLC column. Based on the results obtained from UV-Vis, FT-IR, and HR-MS, the molecule is a polyketide with a molecular formula of C52H78O13 and an exact mass of 911.5490 amu. The partial structure of this compound has been determined using 1D and 2D NMR spectroscopy.

Ces travaux ont pour objet l’élaboration de nouvelles structures poreuses non tissées antibactériennes. Différentes stratégies ont été développées : l’une a consisté à élaborer des mats poreux par electrospinning en utilisant un polymère biosourcé et biocompatible et l’autre voie consistait à modifier un support fibreux provenant d’un masque de protection respiratoire commercial. La méthode des assemblages par interactions ioniques en superposant de façon alternative les couches de polymères cationiques et les polymères anioniques à la surface du filtre médian en polypropylène (PP) a permis d’élaborer de nouvelles structures ayant de bonnes propriétés antioxydantes et antibactériennes. Le polymère anionique, dérivé du polymère de cyclodextrine présente l’avantage de pouvoir encapsuler un agent antimicrobien biosourcé, le carvacrol. Une autre approche a consisté à modifier des supports en PP avec de l'acide tannique, un polyphénol d'origine naturelle. Dans cette étude, deux stratégies ont été mises en place afin de fonctionnaliser le PP avec de l’acide tannique (AT). La première est l’extrusion réactive du PP avec l’AT en présence (ou non) de peroxyde de dicumyle (DCP) pour greffer directement l’acide tannique sur le PP. La deuxième stratégie consiste à polymériser l’AT au travers d’une couche poreuse de PP extraite d’un masque de protection commercial, afin de permettre l’immobilisation physique de l’AT à la surface du mat fibreux en PP. Le greffage en surface via un procédé “grafting from” a également été étudié. Ces matériaux ont montré de bonnes propriétés antiradicalaires. / The purpose of this work is to develop new antibacterial non-woven porous structures. Different strategies were developed: one was to develop porous structures by electrospinning using a biosourced and biocompatible polymer, the other was to modify a fibrous support from a commercial respiratory protection mask. Assembling materials using ionic interactions by alternatively superposing cationic polymer layers and anionic polymers on the surface of the polypropylene (PP) median filter allowed to develop new structures with good antioxidant and antibacterial properties. The anionic polymer, derived from the cyclodextrin polymer, has the advantage of being able to encapsulate a bio-based antimicrobial agent, carvacrol. Another approach was to modify PP filters with tannic acid, a naturally occurring polyphenol. In this study, two strategies were implemented to functionalize PP with tannic acid (TA). The first is the reactive extrusion of PP with TA in the presence (or not) of dicumyl peroxide (DCP) to directly graft tannic acid onto PP. The second strategy consists in polymerizing the TA through a porous layer of PP extracted from a commercial mask, in order to allow the physical immobilization of the TA on the surface of the PP fibrous mat. Surface grafting using a "grafting from" process was also studied. These materials have shown good anti-free radical properties.

Objetivo: Evaluar el límite de inhibición bacteriana del Hidróxido de Calcio en polvo sobre cepas de Enterococcus faecalis ATCC® 29212™. Materiales y Métodos: Este estudio experimental in vitro utilizó 40 placas petri con agar BHI solidificado. Se preparó la solución de Hidróxido de Calcio [Ca(OH)2] en polvo de la marca Biodinámica y Maquira a la concentración de 1g/ml. La Concentración Mínima Inhibitoria (CMI) y la Concentración Mínima Bacteriostática (CMB) fueron evaluadas mediante diferentes concentraciones de Ca(OH)2 (0.02 g/ml - 1g/ml). La CMI fue evaluada mediante la técnica de Kirby Bauer y la CMB mediante la técnica de Recuento de Unidades Formadoras de Colonia (UFC). Resultados: Al evaluar la actividad antibacteriana de Ca(OH)2 contra la cepa de E. faecalis, se observó las siguientes medias del halo de inhibición: Biodinámica 22.66 ± 0.38 mm; Maquira 23.58 ± 0.47 mm y Clorhexidina al 12% (control positivo) 23.93 ± 0.73 mm. La CMI del Ca(OH)2 en polvo de ambas marcas fue de 0.063 g/ml como límite inferior, confirmando la inhibición del crecimiento de la cepa de Enterococcus faecalis ATCC® 29212™, siendo el límite superior de 1g/ml. Por otro lado, el CMB del Ca(OH)2 en polvo muestra un valor de 0.031 g/ml para la marca Biodinámica y 0.063 g/ml. para la marca Maquira. Conclusión: El límite inferior y superior de inhibición bacteriana del Hidróxido de Calcio, sobre cepas de Enterococcus faecalis ATCC® 29212™ fue de 0.063 g/ml y de 1gr/ml, respectivamente. / Objective: To evaluate the bacterial inhibition limit of Calcium Hydroxide powder on strains of Enterococcus faecalis ATCC® 29212™. Materials and Methods: This in vitro experimental study used 40 petri dishes with solidified BHI agar. The solution of Calcium Hydroxide [Ca(OH)2] powder of the brand Biodinámica and Maquira was prepared at the concentration of 1 g / ml. The Minimum Inhibitory Concentration (MIC) and the Minimum Bacteriostatic Concentration (MBC) were evaluated using different concentrations of Ca(OH)2 (0.02 g /ml - 1 g/ml). The CMI was evaluated using the Kirby Bauer technique and the CMB using the Colonies Formation Units (CFU) technique. Results: The antibacterial activity of Ca(OH)2 against the E. faecalis strain, presented the following means of inhibition halo were observed: Biodinámica 22.66 ± 0.38 mm; Maquira 23.58 ± 0.47 mm and 12% Chlorhexidine (positive control) 23.93 ± 0.73 mm. The MIC of the Ca(OH)2 powder of both brands was 0.063 g / ml as the lower limit, confirming the inhibition of the growth of Enterococcus faecalis ATCC ® 29212 ™ strain with the upper limit of 1 g / ml. On the other hand, CMB of the Ca(OH)2 powder shows a value of 0.031 g / ml for Biodinámica and 0.063 g / ml. for Maquira. Conclusion: The lower and upper limit of bacterial inhibition of Calcium Hydroxide, on strains of Enterococcus faecalis ATCC ® 29212™ was 0.063 g / ml and 1 g / ml, respectively. / Tesis

Thesis (M.Sc. (Microbiology)) --University of Limpopo, 2011 / The leaves of fifteen plant species were collected from the Lowveld Botanical Garden in Nelspruit, Mpumalanga Province, South Africa. The collection was based on a list of plants and their ethnopharmacological information provided by the Phytomedicine Programme at the University of Pretoria. The dried leaves of the plants were powdered and extracted using hexane, dichloromethane, acetone and methanol. The extracts were screened for antibacterial activity against Mycobacterium smegmatis and Rhodococcus erythropolis. The acetone extract of Milletia stulhimannii was the most active, showing activity against Mycobacterium smegmatis and Rhodococcus erythropolis with MIC values 0.13 and 0.08 mg/ml, respectively. Acetone extracts for all plants had the lowest MIC values ranging between 0.11-1.25 mg/ml and 0.08-1.25 mg/ml for M. smegmatis and R. erythropolis, respectively. Milletia stulhimannii, Albizia gummifera, Xanthocercis zambesiaca and Barringtonia racemosa extracts have shown the greatest potential for anti-tubercolosis agents. These were all active against M. smegmatis with an average MIC value of acetone extracts of 0.13 mg/ml. Apodytes dimidiata was selected for the isolation of active compounds since its activity on qualitative antibacterial activity assays was highly prominent on TLC plates in comparison to the other plant extracts. Two compounds were isolated from A. dimidiata but after purification, their MICs were above 2.5 mg/ml indicating a possible loss of activity during purification. The preliminary NMR spectra analysis suggested that the compounds were a long fatty acid and a triterpene. Future work is required to elucidate the chemical structures of the latter compounds and to test the activity of these compounds against Mycobacterium tuberculosis. / Department of Water Affairs, and University of Limpopo Research Development and Administration Office

Thesis (M.Sc. (Agriculture)) --University of Limpopo, 2006 / Refer to document / National Research Foundation (NRF) and Cannon Collins Educational Trust Fund of Southern Africa (CCETSA)

Thesis (M.Sc. (Microbiology)) --University of Limpopo, 2015 / Five plant species: Ziziphus mucronata, Senna italica, Lantana camara, Ricinus communis and Lippia javanica, were selected for this study based on their use in traditional medicine. In preliminary screening, crude extracts were prepared using hexane, dichloromethane (dcm), acetone and methanol. Phytochemical profiles on Thin Layer Chromatography plates of the extracts were obtained by developing the plates in mobile phases of varying polarity. Tests for compounds such as tannins, flavonoids, alkaloids, phlobatannins, terpenes, steroids, cardiac glycosides and saponins were carried out. Antibacterial activity of the extracts was carried out using microdilution assay for Minimum Inhibitory Concentration and bioautography against Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis. Antioxidant activity of the extracts was performed using the 2, 2, diphenyl-1-picrylhydrazyl (DPPH) assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay and Phagoburst test were used to investigate the toxic effects and anti-inflammatory activity of the extracts on mouse Raw 264.7 macrophage cells, respectively. The presence of phytochemicals was observed on the chromatograms after the plates were sprayed with vanillin sulphuric acid reagent. The dcm extracts of the plants showed antibacterial activity against the selected bacterial species on the bioautograms. Senna italica and Z. mucronata showed the most activity bands on the bioautograms. Lippia javanica had the lowest MIC average of 0.56 mg/ml. Antioxidant activity was observed in the extracts of L. javanica and R. communis. The extracts promoted proliferation of the mouse macrophage cells Raw 264.7 at concentrations ranging from 0.31 mg/ml and 0.08 mg/ml. Senna italica leaves were selected for isolation of antibacterial compounds. The isolated compound was analysed on 1H and 13C nuclear magnetic resonance (NMR) and Mass Spectrometry (MS) for structural analysis. The structure could not be elucidated due to impurities in the compound but the tentative structure is a branched chain alkane with at least one ether linkage per repeating unit. Therefore the study shows that there are plant components with biological activities against wound infecting bacteria and a single lead compound was identified. / the National Research Foundation