Dissertations / Theses on the topic 'Antibacterial Screening'

Plants from Northern Nigeria with a history of use in both human and veterinary traditional medicine have been investigated in vitro, for antiviral and antibacterial activity and their cytotoxicity against human cells in culture determined. Aqueous extracts were tested against poliovirus, astrovirus, human and Equine herpes simplex virus and canine and bovine parvovirus, using the microtitre plate inhibition test. Most of the extracts were non toxic to cells in culture and had activity against more than one virus at a dose rate of between 100 and 400 g/100. the most effective of the plant extracts were those of Bauhania thonningi, Anacardium occidentale, Boswelia dalzeilii, Dichrostacyls glomerata and Sterculia setigera, which were found to be active against all the viruses tested. In addition, most of the extracts were active against those Gram positive bacteria tested. Two of the plants, Anogeissus schimperi and Anacardium occidentale, had good antibacterial activity against the Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa. Attempts were made to isolate and identify the active components in these extracts with activity against both viruses and bacteria using the gel filtration, ion-exchange chromatography and thin layer chromatography. Activities were associated with fractions identified as flavonoids, alkaloids, terpenoids and polyphenols and were found, either individually or in combination, to have antiviral action. A field trial of the active plant extracts was carried out using cattle with clinical diarrhoea and dysentery and comparing recovery patterns with similar administering extracts to cattle with clinically diagnosed diarrhoea animals treated with a broad spectrum gut-acting antibiotic, neomycin-sulphate. No significant difference (P=0.778) was seen between the extract and the antibiotic in efficacy of treatment.

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.<br>Master of Science

Natural products and their derivatives have and are still used by humans for various health ailments due to their rich sources of drug discovery. New biologically active compounds from natural products play a key role in drug development. Marine sponges and their associated microbes contain a lot of bioactive compounds that are potential for drug development. These compounds produce chemical compounds with useful pharmaceutical properties such as antitumor, anti-infective, anti-inflammatory, and antibacterial properties. The main focus of this project was on the antibacterial activity of six different sponge specimens. The aim was to screen the antibacterial activity of the sponge specimen’s extracts. In order to do so, a Minimum Inhibitory Concentration assay was performed to screen the sponge's antibacterial activity against E. coli and S. aureus. Analytical HPLC was used for separation and Solid Phase Extraction (SPE) was used for determining the effect of salts towards the inhibition of anti-bacterial activity for two selected extracts. Ethanolic extract of Stylissa massa showed antibacterial activity against S. aureus. SPE would be a rapid purification step to remove the salts present in sponges at a high concentration but it has not shown a significant effect on the inhibition of antibacterial activity. However, further separation and purification need to be done to be able to completely screen for all the six different sponge specimens.

Drug-resistant pathogenic and opportunistic bacteria are increasing in occurrence and prevalence, and pose a dangerous threat to human health. In the search for novel antibiotics with which to combat this threat, plants, specifically those used in traditional medicine with ascribed antibacterial properties, offer a promising and potentially vast source of such therapeutic compounds. The purpose of this study was therefore to screen chemical extracts created from various plant species for antibacterial properties versus pathogenic bacterial species. In the course of these antibacterial assays, we successfully identified a methanol extract derived from Artemisia tridentata tridentata plant material as capable of inhibiting the growth of the opportunistic pathogen Staphylococcus aureus. Three sub-fractions were created using hexane, ethyl acetate and water solvents. Each of these extracts displayed significant antibacterial activity versus a wild-type strain over a period of six hours, at concentrations as low as 62.5 µg/ml. The extracts also demonstrated an enhancement of antibiotic effects when combined with ampicillin, G418 sulfate or amikacin, for a period of up to twelve hours. Though the efficacy of the extracts was lessened when tested against an ampicillin-resistant strain, significant enhancement of the efficacy of this antibiotic was still observed. Gas chromatography-mass spectrometry analysis of these three extracts revealed the sesquiterpene lactone achillin as present in each. Column chromatography of the hexane extract resulted in a fraction retaining its antibacterial activity, and still containing this compound, further implicating it as responsible for the antibacterial activity of this plant. The results of serial dilution and plating of extract-treated samples, along with those of ethidium bromide assays and transmission electron microscopy analysis, indicated a bacteriostatic mechanism of action involving disruption of the bacterial membrane, which is in agreement with the literature on the antibacterial properties of this plant, and those of sesquiterpene lactones, respectively. We therefore conclude that achillin, likely produced as a secondary metabolite by Artemisia tridentata tridentata, possesses growth inhibitory properties versus Staphylococcus aureus, and should be isolated and studied further for the purposes of evaluating its potential use, either as a stand-alone antibiotic, or as an adjunctive therapeutic, in the treatment of drug-resistant bacterial pathogens.

>Magister Scientiae - MSc<br>Nanotechnology has emerged as an elementary division of modern science and stemmed directly from green chemistry twelve basic concepts, it receives global attention due to its unique character and ample applications. It also has great potential to mitigate the challenges they face in various fields, especially medical sector. Nanodrugs are increasingly considered as a potential candidate to carry therapeutic agents safely into a targeted compartment in an organ, particular tissue or cell. In this study, twenty (20) Libyan plants were selected and evaluated for their potential to synthesis gold and silver nanoparticles. The screening of the different plant extracts was performed using 96 well plate method at 25 °C and 70 °C. The NPs formation was confirmed and characterized using UV- Vis, DLS, HR-TEM and EDX. A well-defined NPs were obtained at high temperature (70 °C). The Au NPs had an average diameter of 92 nm at 25 °C and 66 nm at 70 °C. The zeta potential values were observed to be negative (-14 to -24) and indicate the stability of the Au NPs. The HR-TEM showed polydispersity, which decreased at higher temperature (70 °C). The stability of Au NPs in nutrient broth prior was conducted as well. All the Au NPs under study showed stability, only minimal changes in the UV-Vis spectra can be observed. Two plant extract viz Pistacia atlantica, Junipers phoenicea showed consistent results and forming stable and smaller NPs compared to others, both of the plant extracts and the corresponding NPs were tested against Streptococcus mutans and showed MIC value ~ 49 g/mL. In case of silver NPs, two plant extracts viz J. phoenicea, Rosmarinus officinalis, showed superior results than the others; both plants produced stable and small Ag NPs. The antibacterial activity against S. mutans demonstrated MIC valus ~ 50 g/mL. The synthesised NPs showed a promising bioactivity for developments of new antibacterial agents against S. mutans strains. Dose-dependent activity was observed for the tested NPs.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-07-07T12:56:32Z No. of bitstreams: 1 arquivototal.pdf: 3524208 bytes, checksum: 1dfa6b7ebfd1711a48733025c4052ee9 (MD5)<br>Made available in DSpace on 2017-07-07T12:56:32Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3524208 bytes, checksum: 1dfa6b7ebfd1711a48733025c4052ee9 (MD5) Previous issue date: 2017-02-13<br>Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq<br>The human body has a large bacterial flora, but when these bacteria, the principle of commensal character, become part of site other than it's natural, they can cause some severe diseases. Researches that are based either on the search of new drugs from plants or on the improvement of phytotherapics are in prominence and continue to play an important role nowadays. In this perspective, the aim was to carry out a phytochemical study of Solanum capsicoides All. fruits to isolate and characterize the chemical substances of this species and to use in silico studies to carry out investigations of new molecules potentially active for methicillin resistant Staphylococcus aureus (MRSA) and Escherichia coli using a database created with secondary metabolites isolated from Solanum genus. The phytochemical study of Solanum capsicoides All. fruits resulted in the isolation of three substances: carpesterol, acetylated glucose and 4-hydroxybenzaldehyde. A review of the literature led to the creation of a database with 421 different secondary metabolites isolated from the Solanum genus. Two databases from CHEMBL were selected. The first one with activity against MRSA and another against E. coli. The compounds were classified according to the pIC50 values to generate and validate the model using "Random Forest"(RF). The structure of six new target proteins against S. aureus obtained from the PDB (Protein database) were used for virtual screening of the based on the receptor structure using docking studies by the Molegro Virtual Docker, reaching to select Solanum database molecules capable of interacting in the binding sites of proteins. The RF prediction model for MRSA obtained a percent accuracy of 81%, area under the Receiver Operating Characteristic (ROC) curve of 0.885, selecting 8 molecules with an active potential above 60%. The prediction model for Escherichia coli obtained an accuracy rate of 88%, area under ROC curve of 0.932, selecting 4 molecules with potential probability above 84%. The study of the coupling of six target enzymes to S. aureus selected an average of 50 molecules from the bank of 421 isolated molecules of the genus Solanum with an ability to interact with on active site of each enzyme. In addition, it was possible to obtain 1 molecule with active potential and interaction capacity with 5 enzymes studied, 7 molecules interacting with 3 enzymes and 6 with 2 enzymes of S. aureus. The rutin, a molecule potentially active in the in silico study for S. aureus and E. coli, together with carpesterol, were tested in vitro against these bacteria. Microbiological tests have shown that carpesterol has no antimicrobial activity for the studied strains, and that the rutin has activity only for E. coli. An interaction study with strains of S. aureus ATCC 25923, a standard strain sensitive to all antibiotics, and SAM-01, a multidrug resistant strain, was designed. There was interaction only between rutin and oxacillin, one of the three antibiotics studied in the interaction, for a strain SAM- 01, reducing the resistance of this strain.<br>O ser humano possui uma vasta flora bacteriana que é comensal, mas quando essas bactérias, a princípio de caráter comensal, passam a fazer parte de outro sítio que não o seu natural, podem causar graves patogenias. Pesquisas que se fundamentam na busca por novos medicamentos a partir de plantas ou no melhoramento de fitoterápicos já existentes vem se destacando e continuam a desempenhar um papel importante nos dias de hoje. Nessa perspectiva objetivou-se realizar um estudo fitoquímico dos frutos de Solanum capsicoides All. para isolar e caracterizar substâncias químicas desta espécie e, utilizando estudos in silico, realizar investigações de novas moléculas potencialmente ativas para Staphylococcus aureus resistente a meticilina (MRSA) e Escherichia coli, utilizando um banco de dados criado com metabólitos secundários isolados do gênero Solanum. O estudo fitoquímico dos frutos de Solanum capsicoides All. resultou no isolamento de três substancias: carpesterol, glicose acetilada e 4-hidroxi-benzaldeído. A revisão de literatura levou à criação de um banco de dados com 421 diferentes metabólitos secundários isolados do gênero Solanum. Foram selecionados dois bancos de dados obtidos a partir do CHEMBL. O primeiro com atividade contra S. aureus multirresistente (MRSA) e o outro contra E. coli. Os compostos foram classificados de acordo com valores de pIC50 para gerar e validar o modelo utilizando “Random Forest”(RF). A estrutura de seis novas proteínas alvo contra S. aureus obtidas do Protein Data Bank (PDB) foram utilizadas para triagem virtual baseada na estrutura do receptor utilizando estudos de “docking” com o software Molegro Virtual Docker, a fim de selecionar moléculas do banco de dados de Solanum com potencial capacidade de interagir nos sítios de ligação dessas proteínas. O modelo RF de predição para S. aureus multirresistente obteve uma porcentagem de acerto de 81%, área sob a curva Receiver Operating Characteristic (ROC) de 0,885, selecionando 8 moléculas com potencial ativo superior a 60%. O modelo de predição para Escherichia coli obteve taxa de acerto de 88%, área sob curva ROC de 0,932, selecionando 4 moléculas com probabilidade de potencial ativo superior a 84%. O estudo do docking das seis enzimas alvo para S. aureus selecionou uma média de 50 moléculas do banco de 421 moléculas isoladas do gênero Solanum com a capacidade de interagir no sitio ativo de cada enzima. Analisando moléculas multitarget, foi possível obter 1 molécula com potencial ativo e capacidade de interação com 5 das 6 enzimas estudadas, 7 moléculas interagindo com 3 enzimas e 6 com 2 enzimas de S. aureus. A rutina, uma molécula potencialmente ativa no estudo in silico para S. aureus e E. coli, juntamente com o carpesterol, foram testadas in vitro contra essas bactérias. Os testes microbiológicos mostraram que o carpesterol não possui atividade antimicrobiana para as cepas estudadas, e que a rutina possui atividade apenas para a cepa de E. coli. Foi realizado ainda estudo de interação com as cepas de S. aureus ATCC 25923, uma cepa padrão sensível a todos os antibióticos, e SAM-01, uma cepa multirresistente. Houve interação apenas entre a rutina e a oxacilina, um dos três antibióticos estudados na interação, para a cepa SAM-01, diminuindo a resistência dessa cepa.

O gênero Salmonella constitui um problema para a saúde animal e humana. Com a finalidade de evitar a ocorrência da transmissão deste microrganismo e a interrupção da doença provocada por eles, deve-se utilizar desinfetantes e antibióticos. A exposição continuada a estes agentes pode resultar no surgimento de resistência microbiana. Tendo em vista este fenômeno, a pesquisa por novos agentes antimicrobianos a partir de extratos vegetais se faz necessária, pois existe a possibilidade de se encontrar substâncias eficazes contra a resistência de micro-organismos já disseminados no ambiente. Os objetivos deste estudo foram: monitorar a resistência das 134 amostras de Salmonella isoladas em produtos de origem animal frente a antibióticos comumente utilizados na rotina veterinária; testar a atividade dos desinfetantes frente às Salmonellas resistentes; testar a hipótese de resistência cruzada entre grupos antibióticos e desinfetantes; avaliar a atividade bactericida de extratos de plantas medicinais nativas no Rio Grande do Sul sobre salmonelas padrão; avaliar a atividade antibacteriana do macerado hidroalcoólico de Achyrocline satureioides frente às salmonelas resistentes; fazer prospecção fitoquímica do macerado hidroalcoólico e do decocto de Achyrocline satureioides, Realizou-se o teste de sensibilidade a antibióticos frente a 134 amostras de Salmonella spp. isoladas em produtos de origem animal. Cinquenta e uma (51) salmonelas foram resistentes a pelo menos um antibiótico. Após, utilizou-se o teste de suspensão da avaliação quantitativa da atividade bactericida de desinfetantes. Foram testados os desinfetantes cloreto de cetil trimetilamônio (amônia quaternária), digluconato de clorexidina (clorexidina), hipoclorito de sódio e iodóforo, em quatro concentrações frente salmonelas resistentes, nos tempos 5, 15 e 60 minutos, densidade populacional 107 UFC/ mL. O iodóforo e o hipoclorito de sódio inativaram os isolados na maior parte dos experimentos. Porém, cloreto de cetil trimetilamônio (amônia quaternária) e digluconato de clorexidina (clorexidina) mesmo nas maiores concentrações do desinfetante, houveram micro-organismos resistentes. Pode-se perceber que não houveram relações concomitantes de resistência entre antibióticos e desinfetantes, levando-se em consideração todas as variáveis testadas. Na triagem inicial da atividade antibacteriana das plantas medicinais, utilizou-se a proporção de 10 g de planta para 100 mL do líquido extrator, em 8 e 24 horas, densidade populacional de Salmonella Choleraesuis (ATCC 10.708) 107, 106 e 105 UFC/ mL e formas decocção e maceração hidroalcoólica das plantas Achyrocline satureioides, Bidens pilosa, Conyza bonariensis, Jacaranda micrantha, Myrciaria cuspidata e Smilax cognata. A. satureioides, como macerado hidroalcoólico apresentou inativação nos tempos 8 e 24 horas, mostrando maior redução logarítmica. A. satureioides no macerado hidroalcoólico foi testada frente a 51 salmonelas resistentes. Observou-se que quanto maior o tempo de contato, nas menores densidades populacionais, maior foi o número de inativações. A atividade antibactericida do macerado hidroalcoólico de A. satureioides sugere o seu uso como ingrediente antisséptico em pomadas para uso animal ou humano, ou ser empregado na imersão de utensílios e partes removíveis de maquinários das indústrias alimentícias, frigoríficos, tendo como objetivo promover a desinfecção destes insumos. A prospecção fitoquímica realizada com o decocto e com o macerado hidroalcoólico de A. satureioides detectou a presença de compostos fenólicos, taninos hidrolisáveis e condensados, flavonóis e saponinas. O decocto de A. satureioides, diferentemente do macerado hidroalcoólico, não apresentou taninos hidrolisáveis e saponinas.<br>The genus Salmonella constitutes a problem for animal and human health. In order to prevent the transmission of this organism and the interruption of the disease caused by them, you should use disinfectants and antibiotics. Continued exposure to these agents can result in the emergence of microbial resistance. In view of this phenomenon, the search for new antimicrobials from plant extracts is necessary, because there is a possibility to find effective substances against the resistance of microorganisms already disseminated in the environment. The objectives of this study were: to monitor the resistance of 134 samples of Salmonella isolated in products of animal origin in front of antibiotics commonly used in veterinary routine; test the activity of disinfectants against the salmonellas resistant; test the hypothesis of cross-resistance between antibiotics and disinfectants; evaluate the bactericidal activity of extracts of medicinal plants native to the Rio Grande do Sul on Salmonella standard; evaluate the antibacterial activity of hydroalcoholic macerate to Achyrocline satureioides to Salmonella resistant; do the hydroalcoholic macerate phytochemical prospecting and to Achyrocline satureioides vegetable water, antibiotic sensitivity testing in front of 134 samples of Salmonella spp. isolated in products of animal origin. Fifty-one (51) Salmonella were resistant to at least one antibiotic. After, we used the suspension test of quantitative evaluation of bactericidal activity of disinfectants. Have been tested the disinfectants cetyl trimetilamônio chloride (quaternary ammonia), chlorhexidine digluconate (chlorhexidine), sodium hypochlorite, iodophor in four concentrations Salmonella resistant front, 5 times, 15 and 60 minutes, population density 107 CFU/ mL. The iodophor and sodium hypochlorite inactivated isolates in most experiments. However, cetyl trimetilamônio chloride (quaternary ammonia) and chlorhexidine digluconate (chlorhexidine) even in the largest concentrations of disinfectant, there were resistant microorganisms. One can notice that there were no concurrent relationships of resistance between antibiotics and disinfectants, taking into account all the variables tested. In the initial screening of antibacterial activity of medicinal plants, the proportion of 10 g of plant for 100 mL of liquid Extractor, in 8 and 12:00 hour, population density of Salmonella Choleraesuis (ATCC 10,708) 107, 106 and 105 CFU/ mL decoction and hydroalcoholic maceration and the plants Achyrocline satureioides, Bidens pilosa, Conyza bonariensis, Jacaranda micrantha, Myrciaria cuspidata and Smilax cognate. A. satureioides hydroalcoholic macerate as presented 8 and 12 hour in inactivation, showing greater logarithmic reduction. A. satureioides in hydroalcoholic macerate has been tested in front of Salmonella resistant 51. It was observed that the longer the time of contact, the smallest population densities, higher number of inactivation. The antibacterial activity of hydroalcoholic macerate of A. satureioides suggests its use as an antiseptic ingredient in ointments for human or animal use, or be employed in the immersion of utensils and removable parts of machinery of food industries, refrigerators, aiming to promote the disinfection of these inputs. The phytochemical held prospecting with the vegetable water and hydroalcoholic macerate of A. satureioides detected the presence of phenolic compounds, hydrolysable and condensed tannins, flavonols and saponins. The vegetable water of A. satureioides, unlike the hydroalcoholic macerate, hydrolysable tannins and failed to provide saponins.

There has been an increase in number of human infections by mycobacteria and opportunistic pathogens of the closely related nocardioform bacteria. Frequent multiple drug resistance in these organisms makes it desirable to identify novel targets for antimicrobial agents. Bacteriophages offer one way to do this as analysis of their DNA reveals great diversity in their genetic makeup, suggesting variety in the way they interfere with host cells. Four novel nocardiophages were therefore isolated from soil and characterized. Libraries of their nucleic acid were constructed and screened for clones inhibitory to a nocardioform of the genus Rhodococcus. Nine clones were characterized, and minimum necessary DNA for inhibitory activity sequenced. Of 18 ORFs predicted on these DNAs, 13 could not be assigned a function. Genes similar to ones in databases apparently interfered with DNA metabolism, protein synthesis, or integrity of plasma membrane. This genetic approach may be an efficient and effective way to discover novel targets for antibiotics.

碩士<br>長庚大學<br>中醫學系天然藥物<br>99<br>Abstract 1 Life finds the way to survive! After the widely clinical applications of the first antibiotics during the end of world war II bacteria from clinical and non-clinical settings have developed resistence soonly and are becoming increasingly multi-resistant to conventional antibiotics. Ten years ago, concerns were focused mainly on Gram-positive bacteria, particularly methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus sp. Recently, more and more clinical microbiologists agree that multi-resistant Gram-negative bacteria pose the greatest risks to public health. Consequently there aries an imperious need to explore and develop new chemotherapeutic antibacterials agents. Thus, we collected, isolated, identified and cultivated 17 cyanobacterial strains (especially terrestrial species) from different habitats in Taiwan. In the preliminary experiment, the cyanobacterial strains will firstly be cultivated in small scale (2 L). The antimicrobial activities of cyanobacterial crude extracts (ethyl acetate- and methanol-soluble fractions) were then assessed using a variety of algal, fungal, and bacterial strains including pathogenic species. The crude extracts from 17 cyanobacterial strains were able to apparently inhibit the growth of Staphylococcus aureus, Candida albicans, Chlorella sp., and Anabaena sp. The significant antibacterial activity of cyanobacteral extrects against Staphylococcus aureus is of special interest. This study then conducted the large scale (50 L) cultivation of cyanobacteral strain YRCC3 which showed the apparent growth inhibitory activity against Staphylococcus aureus and the ability to be incubated easily in the lab with high productivity (14 ± 5 g/L). Cyanobacterial stain YRCC3 belongs to the genus of Anabaena sp. It is reported that a few antimicrobial compounds have been isolated from this genus. In this study, the active compounds in the methnol-soluble extract of strain YRCC3 was separated sequentially by liquid-liquid partition, silica gel open column, gel permeation and HPLC. Abstract 2 Traditional Chinese medicine is one of the most frequently applied alternative therapies in China and Taiwan. Among them, Qing Dai is utilized by traditional Chinese medicine to treat fever, mumps, sore-throat, hepatitis and trauma. In Taiwan, Strobilanthes formosanus (Acanthaceae) is the main source of Qing Dai. However, there were few chemically and pharmacologically investigations to support these traditional claims. Recently, some reports described that the major ingredients of Qing Dai including indigo, indirubin and trptanthrin were manifested about its anti-inflammatory activity. However, the antimicrobial activity of Qing Dai still remained to be studied. In this study, we prepared the ethyl acetate and methanol crude extracts from Qing Dai. The antimicrobial activities of Qing Dai crude extracts were then tested against six strains of fungi and three strains of bacteria. The significant antimicrobial activity of Qing Dai against Staphylococcus aureus is of special interest. So we used Staphylococcus aureus to lead the purification process and obtained seven compounds. Their structures were elucidated using UV, IR, NMR spectroscopy and mass spectrometry. Trptanthrin, one of the major ingredients of Qing Dai, may be the most important compound which is responsible for the growth inhibitive activity to against Staphylococcus aureus.

Thesis (M.Sc. (Microbiology)) --University of Limpopo, 2011<br>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.<br>Department of Water Affairs, and University of Limpopo Research Development and Administration Office

While the ‘war’ on infectious diseases has been considered won, antibiotic-resistant bacteria are currently responsible for 25,000 death’s yearly in Europe, bearing a predicted cost of €1.5 billion. Incidence of health-care associated infections in the United States of America is estimated at 1.7 million per year. No new broad-spectrum antibiotic has been introduced since the 1960s and as of 2011 only 2 agents are in Phase III clinical trials. As the antibiotic pipeline is clearly exhausted, new antibiotics are urgently needed. The present work evaluates Fourier Transform InfraRed Spectroscopy (FTIRS) as the basis of a novel Antibiotic Drug Discovery (ADD) platform. Initially the historical development of antibiotics was presented to contextualize the reader with the state-of-the-art. Resistance was discussed to bridge its upsurge and exhausted ADD platforms. Followed an objective review of infrared spectroscopy’s core concepts, its equipment developments alongside various applications for microbiological studies, the pharmaceutical and food industries besides diagnosis of several diseases. A high-throughput macro-cultivation protocol was adapted to determine minimum inhibitory concentrations of 14 antibiotics on an Escherichia coli model organism, while simultaneously building a spectral library. In order to comprehensively analyze such library a graphic user interface was constructed and used throughout this research. This allowed fair comparison of different pre-processing strategies on two alternative multivariate analysis algorithms, from which an optimal model was created for each proposed objective, in the context of an ADD platform. While some models were short of their objective, overall results were rather positive and suggest, alongside reviewed literature, that FTIRS could, and should, potentiate a new generation of antibacterial drug discovery platform that outperforms current alternatives.<br>Apesar de a ‘guerra’ contra as Doenças Infecciosas já ter sido considerada ganha, bactérias resistentes a antibióticos continuam a causar 25,000 mortes por ano na Europa, com um custo estimado em €1.5 mil milhões. Nos Estados Unidos da América a incidência de infecções nosocomiais foi de 1.7 milhões por ano. A taxa de descoberta de antibióticos é quase nula, o último antibiótico de largo espectro descoberto foi em 1960 e desde 2011 apenas 2 compostos estão na fase III de ensaios clínicos, por isso são urgentemente necessários novos antibióticos. O presente trabalho avalia a espectroscopia de infravermelhos com transformada de Fourier (FTIRS) como base de uma nova plataforma de descoberta de antibióticos (ADD). Apresentou-se o desenvolvimento histórico dos antibióticos para contextualizar o leitor da evolução alcançada. Seguidamente discutiu-se o fenómeno da resistência, relacionando o seu aumento com plataformas de ADD exaustas. Uma revisão objectiva dos principais conceitos da espectroscopia de infravermelhos, o desenvolvimento do respectivo equipamento, as aplicações desta tecnologia em estudos microbiológicos, na indústria farmacêutica e alimentar, e no diagnóstico de diversas doenças. Foi adaptado um protocolo de macro-cultivação de alto-débito (HT) a partir do qual foram determinadas as concentrações minimias inibitórias de 14 antibióticos, num organismo modelo de Escherichia coli, enquanto simultaneamente construiu-se um biblioteca espectral. Por forma a analisar de forma compreensiva essa mesma biblioteca foi montada uma interface gráfica (GUI). Isto permitiu comparar o impacte de diferentes estratégias de pré-processamento em dois algoritmos de análise, a partir dos qual foi determinado um modelo óptimo para cada objectivo proposto, num contexto de ADD. Enquanto que alguns modelos ficaram aquém dos objectivos, os resultados são no geral positivos e sugerem, como a literatura revista, que a FTIRS pode, e deve, potenciar uma nova geração de plataformas de ADD que competem com as alternativas actuais.

This thesis contains six chapters and the research presented within focuses on three parts. The first part (chapter one to three) was the isolation of antibacterial and antioxidant mushroom metabolites, and the synthesis towards analogues of discovered natural products. The following part (chapter four) was to assess antioxidant activities of synthetic compounds and investigate their SAR (structure-activity relationship), by use of a developed DPPH assay. The last part (chapter five to six) was the discovery of an abiotic halogenation reaction among a class of fungal metabolites, azaphilones. A series of their synthetic analogues were applied as model compounds to understand the mechanism of the discovered unusual and facile non-enzymatic nucleophilic halogenation reaction. Chapter one is a review of antioxidants in mushrooms. The components with antioxidant activities discovered from mushrooms were summarised by their chemical structures and representative samples were listed. In addition, the potential mechanisms of those antioxidants were described in this chapter. In chapter two, extracts of 20 mushrooms from PNG (Papua New Guinea) were screened against both Gram-positive and Gram-negative bacteria, six of them were chosen as chemical survey candidates for their strong antibacterial activities. Extracts of 37 PNG mushrooms were screened for their antioxidant using a modified DPPH assay, 14 of them were chosen as chemical survey candidates based on their antioxidant capacities. In chemical examination guided by bioactivity, five natural products including two novel furan fatty acids (2.1-2.2) and three known compounds, i.e. grifolin (2.3), grifolic acid (2.4) and grifolic acid methyl ether (2.5), were isolated from four selected mushroom samples. Chapter three described the synthetic efforts towards analogues of three isolated antioxidant fungal metabolites (2.3-2.5). Inspired by 15 synthetic targets, a short and versatile general synthetic route was established to deliver 40 compounds possessing structures related to the natural products. Based on these structures, a compound library was established for antioxidant assessment in the following SAR research. The synthetic work resulted in the preparation of 33 novel structures. In chapter four, various antioxidant assays and their working mechanisms were reviewed, as well as their corresponding advantages and drawbacks. A developed DPPH assay was optimised to offer an accurate and reproducible antioxidant evaluation for SAR research in the achieved compound library. The results indicated that a structure with an aromatic ring to be allylated with a carbon chain in addition to the presence of phenols, is required to show an antioxidant activity, and the increased length of a carbon chain (more prenyl units) enhances the antioxidant activity. Chapter five focused on naturally occurring organohalogen compounds with biotic origins. The representative halogenated natural products with a variety of bioactivities were categorised following a chemical structural classification as well as a bioactive classification, to showcase both their structural diversity and bioactive variety. The biosynthetic mechanisms of halogenated natural products were reviewed in this chapter. Chapter six detailed the investigation of a novel halogenation reaction that was discovered to be occurring with some azaphilone fungal metabolites, for example the conversion of (+)-deschlorosclerotiorin (6.2) into (+)-sclerotiorin (6.1). Various non-halogenated model compounds with an azaphilone core structure were created to shed light on the authenticity of numerous halogenated azaphilones reported as natural products. The ensuing synthetic work resulted in an efficient general procedure producing the required targets. The following investigation of the halogenation revealed a novel, facile, non-enzymatic nucleophilic reaction and its proposed mechanism is discussed. The synthetic efforts resulted in the creation of 22 novel structures.