Made available in DSpace on 2014-06-11T19:26:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-07-27Bitstream added on 2014-06-13T18:29:22Z : No. of bitstreams: 1 rodrigues_cm_me_botib.pdf: 878460 bytes, checksum: 50adcf0ae69bed0497b25ed84bbbb9d4 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Baseado nas informações geradas após o seqüenciamento do genoma, foi desenvolvido uma série de meios de cultura de composição definida (XDM1, XDM2, XDM3, XDM4 e XDM5). Portanto, um dos objetivos do presente trabalho foi estabelecer a curva de crescimento da X. fastidiosa no meio definido XDM2. Foi utilizada a estirpe 9a5c de X. fastidiosa mantida nesse mesmo meio de cultura. A medida da taxa de crescimento bacteriano foi realizada através de leituras de densidade óptica durante dezesseis dias com intervalos de 48 horas. A avaliação da viabilidade celular foi realizada através da contagem de unidade formadora de colônia (UFC) por diluição seriada. As duas avaliações apresentaram uma alta correlação (R2= 0,91) verificada através de regressão exponencial. O início da fase estacionária foi obtido após 10 dias de crescimento. De posse dos dados de UFC, foi possível calcular o tempo de geração da X. fastidiosa no meio XDM2, de aproximadamente 21 horas. Comparando o tempo de geração obtido na curva de crescimento em meio PW (11,37h), foi comprovado que a bactéria apresenta um crescimento mais rápido em meio PW do que no meio definido XDM2. Outro aspecto interessante revelado no estudo do genoma funcional da X. fastidiosa está relacionado à expressão de genes associados à patogenicidade. O principal mecanismo de patogenicidade é a formação de biofilme no xilema da planta hospedeira conduzindo ao bloqueio dos vasos do xilema. Análise da expressão diferencial de genes desse fitopatógeno em condição de virulência e durante a formação do biofilme revela padrões de genes associados à adaptação e competitividade no ambiente do hospedeiro. Esses genes possivelmente são ativados no biofilme maduro e são essenciais para sua manutenção na planta. Além disso, sabe-se que bactérias que formam biofilme apresentam resistência crescente a compostos... / Based on informations generated after the genome sequencing, a series of culture media of defined composition were developed (XDM1, XDM2, XDM3, XDM4 e XDM5). Therefore, one of the objectives of the present work was to establish the growth curve of X. fastidiosa in the defined medium XDM2. For this, we utilized the 9a5c strain of X. fastidiosa maintained in this medium. The measurement of the bacterial growth rate was done by optical density analysis during sixteen days with intervals of 48 hours. The evaluation of cellular viability was carried out counting the colony units forming (CFU) by serial dilution. Both evaluations presented a high correlation (R2 = 0,91) verified by exponential regression. The beginning of the stationary phase was observed after ten days of growth. With the data of CFU, it was possible to calculate the generation time of X. fastidiosa in XDM2 medium, estimated to be of approximated 21 hours. Comparing the generation time obtained for the growth curve in PW medium (11,37 hours), we proved that the bacteria grow faster in PW compared with the defined medium XDM2. Another interesting aspect reveled by the study of functional genome of X. fastidiosa is related to the expression of genes associated with pathogenicity. The main mechanism of its pathogenicity is the formation of a biofilm in the xylem of the host plant, causing the blockage of the xylem vessels. Analysis of the differential expression of genes of this phytophatogen in condition of virulence and during the biofilm formation reveals standards associated to adaptation and competition in the host environment. These genes are possibly activated in the mature biofilm and are essential for its maintenance in the plant. Besides, it is known that bacteria that form biofilm present elevated resistance to antimicrobial compounds while biofilm is structured, like antibiotics, heavy metals... (Complete abstract click electronic access below)

The presence of wide spectrum antimicrobial agents triclosan (TCS) and triclocarban (TCC) in the environment has become a concern due to the adverse impact on the environment. Montmorillonite, a widely distributed clay mineral in the natural environment, has been used as an obstacle in landfills to avoid contamination of soil and water by contaminants in leachates due to its high surface area, cation exchange capacity, and abundance. The research reported here focuses on understanding the abiotic oxidative transformation of TCS and TCC by Fe(III)-modified montmorillonite. The overall objectives of this work were: 1) to investigate TCS and TCC oxidative transformation kinetics and transformation products in different environmental conditions, and 2) to elucidate their reaction pathways. TCS was reacted with Fe(III)-modified montmorillonite under the following experimental conditions: 1) at 40% relative humidity and room temperature for up to 100 d with and without UV light exposure; and 2) in aqueous environment with different initial TCS concentrations, light exposure, pH levels, and in the presence of natural organic matters. Reaction in the presence of Na- montmorillonite was conducted for comparison with results from TCS reaction in the presence of Fe(III)-modified montmorillonite. In addition, transformation of TCS in the presence of other types of minerals was also investigated. Transformation of TCC on Fe(III) and Na-montmorillonite in an aqueous environment with and without exposure to light was also studied at different initial TCC concentrations. TCS and TCC transformation products were a) characterized using LC/MS, GC/MS, and computational modeling, and b) quantified using HPLC/UV and GC/MS. The main TCS transformation products were 2,4-dichlorophenol, 2,4-dichlorophenol dimer, chlorophenoxy phenols and, TCS dimers and trimers. 2,8-dichlorodibenzo-p-dioxin was identified under UV light and the sun simulator experiments. Formation of 4-chloroaniline and 3,4-dichloroaniline were confirmed as transformation products of TCC. To the best of our knowledge, this is the first time that 4-chloroaniline and 3,4-dichloroaniline were confirmed as abiotic transformation of TCC. This research has generated a better understanding of the abiotic environmental fate of TCS and TCC and demonstrates the feasibility of utilizing Fe(III)-modified montmorillonite as remediation material for TCS, TCC and other related pharmaceutical and personal care products (PPCPs).

Master of Science / Department of Diagnostic Medicine/Pathobiology / Sanjeev K. Narayanan / Antibiotics are substances produced by bacteria or fungi that are inhibitory to other bacteria and fungi. Antimicrobial compounds include substances that are naturally produced, chemically modified or completely synthetic (chemically designed or synthesized). The chemical modification of naturally produced antibiotic generally results in increase stability, solubility, increased spectrum of activity, or efficacy. Antimicrobial compounds are used in animals to treat and control infectious diseases, and also for growth promotion. Bacteria may gain resistance to antibacterial agents via a variety of mechanisms. There is growing evidence that antimicrobial resistance has significant public health consequences. Rationale use of antimicrobial drugs using appropriate medication at the proper dosage and for duration is one of the important means to reduce selective pressure that helps reduce life of resistant organism. It is also vital to reduce the spread of multi drug resistant organisms in the environment especially in health care facilities. Bacteria evolve rapidly not only by mutation, but also by horizontal gene transfer through the transformation, transduction, and conjugation. Conjugation involves a close contact between two bacteria and transfer of the plasmid that carry many genetic elements. The pathogenic bacteria have the ability to sense as well as respond to the stress in the recipient. The epinephrine and norepinephrine play a key role in stress situations in animals. A previous study showed that norepinephrine (NE), a catecholamine at physiological concentrations promoted the conjugation efficiencies of a conjugative plasmid from a clinical strain of Salmonella typhimurium to an E. coli recipient in vitro. The objective of this study was to determine the effect of norepinephrine on conjugation of two E. coli strains. Both filter mating and liquid mating assays were used. The results revealed that there was no significance difference between the presence and the absence of norepinephrine on conjugative transfer of RP4 plasmid between E. coli strains (FS1290 and C600N) either in filter mating or liquid mating. Further studies are needed to determine whether higher concentration of (more than 20 mM) has any effects on conjugation in E. coli.

Resumo: Baseado nas informações geradas após o seqüenciamento do genoma, foi desenvolvido uma série de meios de cultura de composição definida (XDM1, XDM2, XDM3, XDM4 e XDM5). Portanto, um dos objetivos do presente trabalho foi estabelecer a curva de crescimento da X. fastidiosa no meio definido XDM2. Foi utilizada a estirpe 9a5c de X. fastidiosa mantida nesse mesmo meio de cultura. A medida da taxa de crescimento bacteriano foi realizada através de leituras de densidade óptica durante dezesseis dias com intervalos de 48 horas. A avaliação da viabilidade celular foi realizada através da contagem de unidade formadora de colônia (UFC) por diluição seriada. As duas avaliações apresentaram uma alta correlação (R2= 0,91) verificada através de regressão exponencial. O início da fase estacionária foi obtido após 10 dias de crescimento. De posse dos dados de UFC, foi possível calcular o tempo de geração da X. fastidiosa no meio XDM2, de aproximadamente 21 horas. Comparando o tempo de geração obtido na curva de crescimento em meio PW (11,37h), foi comprovado que a bactéria apresenta um crescimento mais rápido em meio PW do que no meio definido XDM2. Outro aspecto interessante revelado no estudo do genoma funcional da X. fastidiosa está relacionado à expressão de genes associados à patogenicidade. O principal mecanismo de patogenicidade é a formação de biofilme no xilema da planta hospedeira conduzindo ao bloqueio dos vasos do xilema. Análise da expressão diferencial de genes desse fitopatógeno em condição de virulência e durante a formação do biofilme revela padrões de genes associados à adaptação e competitividade no ambiente do hospedeiro. Esses genes possivelmente são ativados no biofilme maduro e são essenciais para sua manutenção na planta. Além disso, sabe-se que bactérias que formam biofilme apresentam resistência crescente a compostos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Based on informations generated after the genome sequencing, a series of culture media of defined composition were developed (XDM1, XDM2, XDM3, XDM4 e XDM5). Therefore, one of the objectives of the present work was to establish the growth curve of X. fastidiosa in the defined medium XDM2. For this, we utilized the 9a5c strain of X. fastidiosa maintained in this medium. The measurement of the bacterial growth rate was done by optical density analysis during sixteen days with intervals of 48 hours. The evaluation of cellular viability was carried out counting the colony units forming (CFU) by serial dilution. Both evaluations presented a high correlation (R2 = 0,91) verified by exponential regression. The beginning of the stationary phase was observed after ten days of growth. With the data of CFU, it was possible to calculate the generation time of X. fastidiosa in XDM2 medium, estimated to be of approximated 21 hours. Comparing the generation time obtained for the growth curve in PW medium (11,37 hours), we proved that the bacteria grow faster in PW compared with the defined medium XDM2. Another interesting aspect reveled by the study of functional genome of X. fastidiosa is related to the expression of genes associated with pathogenicity. The main mechanism of its pathogenicity is the formation of a biofilm in the xylem of the host plant, causing the blockage of the xylem vessels. Analysis of the differential expression of genes of this phytophatogen in condition of virulence and during the biofilm formation reveals standards associated to adaptation and competition in the host environment. These genes are possibly activated in the mature biofilm and are essential for its maintenance in the plant. Besides, it is known that bacteria that form biofilm present elevated resistance to antimicrobial compounds while biofilm is structured, like antibiotics, heavy metals... (Complete abstract click electronic access below) / Orientador: Marcos Antonio Machado / Coorientador: Alessandra Alves de Souza / Banca: Ivan Maia de Godoy / Banca: Henrique Ferreira / Mestre

No description available.

As infecções causadas por patógenos intestinais gram-negativos são importantes fontes de prejuízos na criação de animais domésticos como bovinos, ovinos, suínos e aves. Em muitos casos, opta-se pela administração de antibióticos de amplo espectro para prevenção destas infecções e atenuação das perdas. No entanto, esta prática apresenta risco para a saúde humana, bem como contribui para seleção de cepas resistentes. Nos últimos anos, muito tem se discutido a respeito de novas alternativas para atenuar os prejuízos mencionados sem apresentar o mesmo risco. Uma destas alternativas é a utilização de micro-organismos que apresentam propriedades probióticas que sintetizam substâncias inibitórias sobre espécies de patógenos intestinais. Esta abordagem praticamente elimina o risco de desenvolvimento de resistência aos antibióticos de uso clínico, além de evitar a presença de resíduos nos produtos de origem animal. O termo \"probiótico\" é utilizado atualmente para definir micro-organismos que, administrados em doses e frequências adequadas, conferem benefícios à saúde do hospedeiro. Nos últimos anos, diversos estudos têm sido desenvolvidos envolvendo quatro cepas de Lactobacillus (L. acidophilus ATCC 4356, L. casei ATCC 7469, L. fermentum ATCC 9338 e L. plantarum ATCC 8014) com resultados promissores no tocante às suas características probióticas. Desta forma, a proposta deste estudo foi avaliar a capacidade destas cepas em relação à produção de substâncias que apresentam atividade inibitória sobre espécies patogênicas, intestinais gram-negativas, especificamente Escherichia coli 0112, Escherichia coli 0124, Escherichia coli 0127, Shigella sonnei ATCC 25931, Shigella dysenteriae ATCC 13313, Salmonella enteritidis ATCC 13076. Para tanto, foi avaliada a atividade inibitória de substâncias presentes no sobrenadante livre de células de cada cepa. Adicionalmente, foi realizada a caracterização presuntiva das substâncias responsáveis pela inibição do crescimento microbiano quando os respectivos sobrenadantes foram submetidos a diferentes tratamentos (catalase, enzimas proteolíticas, tratamento térmico e neutralização do pH). A estratégia adotada consistiu na avaliação do crescimento, determinado por turbidimetria, das cepas patogênicas na presença do sobrenadante in natura e tratado. Os valores de absorbância foram analisados estatisticamente pelos testes de ANOVA e Tukey e os resultados mostraram que os sobrenadantes in natura de L. acidophilus ATCC 4356 L. casei ATCC 7469 e L. plantarum ATCC 8014 foram capazes de inibir 5 das 6 cepas dos patógenos intestinais estudadas em níveis de inibição variando de 23% a 53%, sendo que a cepa S. dysenteriae ATCC 13313 não teve seu crescimento inibido pelas cepas de Lactobacillus avaliadas. Demonstrou-se também que apenas o sobrenadante in natura de L. fermentumATCC 9338 apresentou atividade inibitória sobre esta cepa, cujo percentual de inibição variou entre 20% e 35% e entre 36% e 65% para as demais cepas patogênicas. A avaliação dos resultados relativos ao crescimento das cepas patogênicas na presença dos sobrenadantes in natura e tratados revelou que o efeito de inibição observado foi devido a presença de ácidos orgânicos que provocou o abaixamento do pH. Demonstrou-se ainda a ausência de substâncias peptídicas e termolábeis ativas contra as cepas patogênicas avaliadas, bem como peróxido de hidrogênio nos respectivos sobrenadantes. / Infectious diseases caused by gram-negative pathogens are important sources of losses in livestock, especially bovine, ovine and poultry. In many cases, subclinical administration of broad-spectrum antibiotics is chosen as an approach for the prevention of these infections, consequently decreasing these losses. However, this practice presents an important risk to human health, as well as it contributes to the selection of bacterial strains which are resistant to antibiotics usually employed in clinical practice. Therefore, special attention has been given to finding alternative procedures to decrease those losses while eliminating that risk. One of these alternatives consists of using microorganism species which present probiotic properties such as synthesis of inhibitory compounds that act on intestinal pathogen species. This alternative virtually eliminates the risk of developing resistance to broad-spectrum antibiotics, as well as avoids the presence of antibiotic residues in animal products. The term \"probiotic\" is currently used to define microorganism species which promote several benefits to the host, once they are administered frequently and in adequate amounts. In the last few years, several works have been carried out using four Lactobacillus strains (L.acidophilus ATCC 4356, L.casei ATCC 7469, L. fermentum ATCC 9338 and L. plantarum ATCC 8014), and the results have been satisfactory regarding to their probiotic characteristics. Therefore, the aim of this study was to evaluate the ability of these strains to produce inhibitory compounds which are active on gram-negative intestinal pathogen species, specifically Escherichia coli 0112, Escherichia coli 0124, Escherichia coli 0127, Shigella sonnei ATCC 25931, Shigella dysenteriae ATCC 13313 and Salmonella enteritidis ATCC 13076. So, antimicrobial activity of the cell-free supernatant of each strain was evaluated. Additionally, presumptive characterization of these compounds was undertaken by submitting the supernatants to different treatments (catalase, proteolytic enzymes, thermic treatments, pH neutralizing). The strategy consisted of evaluating the growth, estimated by turbidimetry, of the mentioned pathogenic strains in the presence of the original supernatants, as well as in the presence of treated supernatants. Aborbance values were statistically analyzed by means of ANOVA and Tukey\'s test. The results showed that the original supernatants of L. acidophilus ATCC 4356 L. casei ATCC 7469 and L. plantarum ATCC 8014 were capable of inhibiting five of six strains of enteric pathogens at levels varying from 23% to 53%. S. dysenteriae ATCC 13313 was not inhibited by the Lactobacillus strains evaluated. It was also demonstrated that only the original supernatant of L. fermentum ATCC 9338 showed inhibitory activity upon this strain varing from 15% to 32%, and between 36% and 65% regarding to the other strains. Growth evaluation of the pathogenic strains in the presence of the treated and original supernatants revealed that the inhibition effect observed occurred due to the presence of organic acids, which lowered the pH of the supernatants. It was also demonstrated the absence of hydrogen peroxide, peptidic and thermolabile compounds in the supernatants.

As infecções causadas por patógenos intestinais gram-negativos são importantes fontes de prejuízos na criação de animais domésticos como bovinos, ovinos, suínos e aves. Em muitos casos, opta-se pela administração de antibióticos de amplo espectro para prevenção destas infecções e atenuação das perdas. No entanto, esta prática apresenta risco para a saúde humana, bem como contribui para seleção de cepas resistentes. Nos últimos anos, muito tem se discutido a respeito de novas alternativas para atenuar os prejuízos mencionados sem apresentar o mesmo risco. Uma destas alternativas é a utilização de micro-organismos que apresentam propriedades probióticas que sintetizam substâncias inibitórias sobre espécies de patógenos intestinais. Esta abordagem praticamente elimina o risco de desenvolvimento de resistência aos antibióticos de uso clínico, além de evitar a presença de resíduos nos produtos de origem animal. O termo \"probiótico\" é utilizado atualmente para definir micro-organismos que, administrados em doses e frequências adequadas, conferem benefícios à saúde do hospedeiro. Nos últimos anos, diversos estudos têm sido desenvolvidos envolvendo quatro cepas de Lactobacillus (L. acidophilus ATCC 4356, L. casei ATCC 7469, L. fermentum ATCC 9338 e L. plantarum ATCC 8014) com resultados promissores no tocante às suas características probióticas. Desta forma, a proposta deste estudo foi avaliar a capacidade destas cepas em relação à produção de substâncias que apresentam atividade inibitória sobre espécies patogênicas, intestinais gram-negativas, especificamente Escherichia coli 0112, Escherichia coli 0124, Escherichia coli 0127, Shigella sonnei ATCC 25931, Shigella dysenteriae ATCC 13313, Salmonella enteritidis ATCC 13076. Para tanto, foi avaliada a atividade inibitória de substâncias presentes no sobrenadante livre de células de cada cepa. Adicionalmente, foi realizada a caracterização presuntiva das substâncias responsáveis pela inibição do crescimento microbiano quando os respectivos sobrenadantes foram submetidos a diferentes tratamentos (catalase, enzimas proteolíticas, tratamento térmico e neutralização do pH). A estratégia adotada consistiu na avaliação do crescimento, determinado por turbidimetria, das cepas patogênicas na presença do sobrenadante in natura e tratado. Os valores de absorbância foram analisados estatisticamente pelos testes de ANOVA e Tukey e os resultados mostraram que os sobrenadantes in natura de L. acidophilus ATCC 4356 L. casei ATCC 7469 e L. plantarum ATCC 8014 foram capazes de inibir 5 das 6 cepas dos patógenos intestinais estudadas em níveis de inibição variando de 23% a 53%, sendo que a cepa S. dysenteriae ATCC 13313 não teve seu crescimento inibido pelas cepas de Lactobacillus avaliadas. Demonstrou-se também que apenas o sobrenadante in natura de L. fermentumATCC 9338 apresentou atividade inibitória sobre esta cepa, cujo percentual de inibição variou entre 20% e 35% e entre 36% e 65% para as demais cepas patogênicas. A avaliação dos resultados relativos ao crescimento das cepas patogênicas na presença dos sobrenadantes in natura e tratados revelou que o efeito de inibição observado foi devido a presença de ácidos orgânicos que provocou o abaixamento do pH. Demonstrou-se ainda a ausência de substâncias peptídicas e termolábeis ativas contra as cepas patogênicas avaliadas, bem como peróxido de hidrogênio nos respectivos sobrenadantes. / Infectious diseases caused by gram-negative pathogens are important sources of losses in livestock, especially bovine, ovine and poultry. In many cases, subclinical administration of broad-spectrum antibiotics is chosen as an approach for the prevention of these infections, consequently decreasing these losses. However, this practice presents an important risk to human health, as well as it contributes to the selection of bacterial strains which are resistant to antibiotics usually employed in clinical practice. Therefore, special attention has been given to finding alternative procedures to decrease those losses while eliminating that risk. One of these alternatives consists of using microorganism species which present probiotic properties such as synthesis of inhibitory compounds that act on intestinal pathogen species. This alternative virtually eliminates the risk of developing resistance to broad-spectrum antibiotics, as well as avoids the presence of antibiotic residues in animal products. The term \"probiotic\" is currently used to define microorganism species which promote several benefits to the host, once they are administered frequently and in adequate amounts. In the last few years, several works have been carried out using four Lactobacillus strains (L.acidophilus ATCC 4356, L.casei ATCC 7469, L. fermentum ATCC 9338 and L. plantarum ATCC 8014), and the results have been satisfactory regarding to their probiotic characteristics. Therefore, the aim of this study was to evaluate the ability of these strains to produce inhibitory compounds which are active on gram-negative intestinal pathogen species, specifically Escherichia coli 0112, Escherichia coli 0124, Escherichia coli 0127, Shigella sonnei ATCC 25931, Shigella dysenteriae ATCC 13313 and Salmonella enteritidis ATCC 13076. So, antimicrobial activity of the cell-free supernatant of each strain was evaluated. Additionally, presumptive characterization of these compounds was undertaken by submitting the supernatants to different treatments (catalase, proteolytic enzymes, thermic treatments, pH neutralizing). The strategy consisted of evaluating the growth, estimated by turbidimetry, of the mentioned pathogenic strains in the presence of the original supernatants, as well as in the presence of treated supernatants. Aborbance values were statistically analyzed by means of ANOVA and Tukey\'s test. The results showed that the original supernatants of L. acidophilus ATCC 4356 L. casei ATCC 7469 and L. plantarum ATCC 8014 were capable of inhibiting five of six strains of enteric pathogens at levels varying from 23% to 53%. S. dysenteriae ATCC 13313 was not inhibited by the Lactobacillus strains evaluated. It was also demonstrated that only the original supernatant of L. fermentum ATCC 9338 showed inhibitory activity upon this strain varing from 15% to 32%, and between 36% and 65% regarding to the other strains. Growth evaluation of the pathogenic strains in the presence of the treated and original supernatants revealed that the inhibition effect observed occurred due to the presence of organic acids, which lowered the pH of the supernatants. It was also demonstrated the absence of hydrogen peroxide, peptidic and thermolabile compounds in the supernatants.

The objective of this study is to evaluate pseudo-substrates of pantothenate kinase (PanK) for the therapeutic treatment of multidrug resistant bacterial infections of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Pantothenate (Pan) analogs, including N- pentylpantothenamide (N5-Pan) and N-heptylpantothenamide (N7-Pan), hamper bacterial growth by utilizing the PanK enzymes, which normally catalyze the rate determining step of the Coenzyme A biosynthetic pathway. Here we report the structures of SaPanK, Human PanK3 and EcPanK complexed with N7-Pan or N5-Pan, all of which have provided the opportunity to investigate the structural differences of bacterial and human Pan binding sites. The MTT assay showed these analogs to exhibit no apparent cytotoxicity against Human A549 lung adenocarcinoma cells, human HepG2 hepatoma cells and human umbilical vein endothelial cells (HUVEC). The presented structural differences have the potential for aiding the development of species-specific antimicrobial compounds with minimal effects on human cells.

Fresh produce is increasingly implicated in outbreaks of foodborne illness. Internalization of bacterial pathogens into produce is of particular concern as internalized pathogens are unlikely to be removed by surface sanitizers. It is therefore necessary to develop treatments that will reduce their prevalence and numbers on fresh produce. Irradiation is a penetrating nonthermal treatment that effectively eliminates bacteria. Irradiated baby spinach leaves up to 1.0 kGy showed negligible (P>0.05) changes in color, texture, vitamin C, total carotenoids, and chlorophyll content compared to non-irradiated controls throughout storage (15 days at 4oC). This research also shows that irradiation effectively reduces viable Escherichia coli cells internalized in lettuce, and that decontamination is not influenced (P>0.05) by lettuce variety. Irradiation effectively reduced the population of internalized pathogens in a dose-dependent manner (3-4 log reduction at 1.0 kGy). Microscopy images suggest that the contamination sites of pathogens in leafy vegetables are mainly localized on crevices and in the stomata. A careful design of the treatment (understanding dose distribution) will effectively eliminate pathogens while maintaining produce quality. The use of modified atmosphere packaging increased (P<0.05) the sensitivity of pathogens (Salmonella spp. and Listeria spp.) to irradiation in baby spinach leaves (up to 25%). Increasing concentration of oxygen increased (P<0.05) sensitivity of both microorganisms. Radiosensitization could be affected (P<0.05) by production of ozone, which increases with increasing dose-rate and oxygen concentration, and reducing temperatures. Antimicrobial effectiveness of various active compounds was determined against Salmonella spp. and Listeria spp. Inclusion complexes were prepared with antimicrobial compounds and -cyclodextrin. The effectiveness of the microencapsulated compounds was tested by spraying them on the surface of baby spinach leaves inoculated with Salmonella spp. The increase in radiosensitivity (up to 40%) varied with the antimicrobial compound. Spherical poly (DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped eugenol and trans-cinnamaldehyde were synthesized for future antimicrobial delivery applications. All loaded nanoparticles proved to be efficient in inhibiting growth of Salmonella spp. and Listeria spp. The entrapment efficiency for eugenol and trans-cinnamaldehyde was 98% and 92%, respectively. Controlled release experiments (in vitro at 37oC for 72 hrs) showed an initial burst followed by a slower release rate of the antimicrobial entrapped inside the PLGA matrix.

The objective of this study is to evaluate pseudo-substrates of pantothenate kinase (PanK) for the therapeutic treatment of multidrug resistant bacterial infections of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). Pantothenate (Pan) analogs, including N- pentylpantothenamide (N5-Pan) and N-heptylpantothenamide (N7-Pan), hamper bacterial growth by utilizing the PanK enzymes, which normally catalyze the rate determining step of the Coenzyme A biosynthetic pathway. Here we report the structures of SaPanK, Human PanK3 and EcPanK complexed with N7-Pan or N5-Pan, all of which have provided the opportunity to investigate the structural differences of bacterial and human Pan binding sites. The MTT assay showed these analogs to exhibit no apparent cytotoxicity against Human A549 lung adenocarcinoma cells, human HepG2 hepatoma cells and human umbilical vein endothelial cells (HUVEC). The presented structural differences have the potential for aiding the development of species-specific antimicrobial compounds with minimal effects on human cells.