Alfalfa sprouts have been implicated in several salmonellosis outbreaks in recent years. Electrolysis of a NaCl solution generates, in the anode side of a two-chamber electrolyzer, acidic electrolyzed oxidizing (EO) water which contains Cl₂, HOC1, dissolved oxygen, and it is characterized by a low pH and a high oxidation reduction potential (ORP). The disinfectant effects of EO water against Salmonella enterica both in an aqueous system and on artificially contaminated alfalfa seeds were determined. EO water (pH 2.54 to 2.38, and ORP +1083 to +1092 mV) exhibited strong potential for the inactivation of S. enterica in an aqueous system (producing a reduction of at least 6.6 log CFU/ml). Treatment of artificially contaminated alfalfa seeds with EO water at a seed-to-EO water ratio of 1:100 for 15 and 60 min significantly reduced Salmonella populations by 2.04 and 1.96 log CFU/g, respectively (P < 0.05), while a Butterfield's buffer wash decreased Salmonella populations by 0.18 and 0.23 log CFU/g, respectively. Germination of seeds was not significantly affected (P > 0.05) by a treatment for up to 60 min in electrolyzed water. Further studies were undertaken to investigate whether •OH radical species are present in EO water or are formed when EO water reacts with iron ions. Electron spin resonance spectroscopy (ESR) coupled with the spin trapping technique was used for the detection of free radicals. The DMPO-OH spin adduct, characteristic to •OH radicals, was not observed. In the presence of DMPO (5,5-dimethyl-l-pyrroline-N-oxide), the reactions of EO water with Fe³⁺ and Fe²⁺ yielded the spin adduct DMPO-OH. However, the addition of •OH radical scavengers (ethanol, methanol, and mannitol) to the reaction mixture, did not reduce the DMPO-OH signal nor generated the characteristic DMPO-alkyl spin adducts. This indicated that the DMPO-OH signal was due to a nucleophilic addition of water to DMPO and not to trapping of OH radicals. Addition of DMPO to EO water, generated an ESR spectrum identical with that of 5,5-dimethyl-2-pyrrolidone- N-oxyl (DMPOX). Hypochlorous acid, generated in the anode chamber, is proposed to oxidize the spin trap DMPO with the formation of DMPOX. / Graduation date: 2004

Thesis (M. Ag. Sc.)--University of Adelaide, Dept. of Horticulture, Viticulture and Oenology, 1997. / Includes bibliographical references (leaves 184-195).

A survey of the quality of brassica seed used for commercial transplant production revealed that overall germination was high but that there was a wide range of seed vigour, as assessed by the controlled deterioration test. These vigour differences were reflected in the performance of seed in modules, the use of low quality seed resulting in problems of a reduced rate of emergence and a lack of seedling uniformity. The improvement of seed quality was considered using three approaches. First, the use of seed soakwater conductivity as a means of predicting germination was investigated for individual seeds. This proved not to be a reliable means of sorting seeds. However, the combination of controlled deterioration followed by assessment of conductivity on bulks of seeds gave a good indication of vigour and was developed as a possible rapid vigour test. Secondly, the relation between seed size and seed performance was examined. No clear relationship was found between seed size and seed quality, although, seed size had a marked effect on seedling growth, the larger seeds consistently producing taller seedlings. Thirdly, a physiological means of seed improvement was considered. Aerated hydration in water columns was successful as an invigoration treatment resulting in increased germination rate, root length of seedlings and seed vigour leading to enhanced performance in modules. These improvements were maintained after drying and storage. The optimum short-term treatment was 8 hours hydration at 25oC. The improvement could be partly attributed to germination advancement and in addition the effect of temperature, aeration and the greater improvement of aged seeds suggested that repair processes were activated during aerated hydration. Also, prolonged hydration for up to 32 hours gave an improvement such that the performance of aged seed was not significantly different to unaged seed. Finally, one possible mechanism by which aerated hydration resulted in such improvements in seed quality was investigated. DNA synthesis began approximately 24 hours earlier in unaged than in aged seed. Hydration reduced this lag phase, indicating that the onset of DNA replication was accelerated. The use of hydroxyurea, an inhibitor of DNA replication, indicated that the repair of accumulated DNA damage may have been occurring between the 16th and 32nd hours of germination.

Thesis (Ing. Agrónomo)--Facultad de Ciencias Agrícolas Pecuarias y Veterinarias, Universidad Técnica de Oruro. / Reproduced from copy at BYU's Benson Institute. Includes bibliographical references (leaves 121-123).

The focus of my research was on the secondary metabolites produced by crucifer plants under stress and their biological activity against fungi. Both cultivated and wild plants were investigated to isolate phytoalexins and phytoanticipins, and determine their metabolite profiles.<p>The first chapter of this thesis describes cruciferous plants and their most important pathogenic fungi. These plants are divided into three groups: oilseeds, vegetables and wild species. The metabolites isolated from these plants and their biosynthetic studies are reviewed. In addition economically important necrotrophic fungi such as <i>Leptosphaeria maculans</i>, <i>Alternaria brassicae</i>, <i>Sclerotinia sclerotiorum</i> and <i>Rhizoctonia solani</i> are also reviewed along with their phytotoxins. <p>The second chapter of this thesis describes the detection, isolation, structure determination, syntheses of stress metabolites and biological activity of these metabolites against <i>L. maculans</i>, <i>S. sclerotiorum</i> and <i>R. solani</i>. The investigation of cauliflower led to the isolation of seven phytoalexins: 1-methoxybrassitin (55), spirobrassinin (71), isalexin (64), brassicanal C (60), caulilexins A (106), B (107), and C (105). The phytoalexins caulilexins A (106), B (107) and C (105) were reported for the first time. Caulilexin A (106), having a disulfide bridge, showed the highest activity against S. sclerotiorum and R. solani among the known phytoalexins. Similarly four phytoalexins: 1-methoxybrassitin, brussalexins A (121), B (117) and C (118) along with four metabolites: ascorbigen (51), diindolylmethane (50), 1-methoxy-3,3-diindolylmethane (119) and di-(1-methoxy-3-indolyl)methane (120) were isolated from Brussels sprouts. The phytoalexins brussalexins A (121), B (117) and C (118) are new metabolites. Brussalexin A (121) is the only cruciferous phytoalexins having an allyl thiolcarbamate functional group. The metabolite 1-methoxy-3,3-diindolylmethane (119) is reported for the first time.<p>The investigation of brown mustard for polar metabolites led to the isolation of indole-3-acetonitrile (76) and spirobrassinin (71) along with isorhamnetin-3,7-diglucoside (134). Investigation of wild species such as Asian mustard, sand rocket, wallrocket, hedge mustard and Abyssinian mustard for production of stress metabolites led to the isolation of indole-3-acetonitrile (76), arvelexin (84), 1,4-dimethoxyindole-3-acetonitrile (137), rapalexins A (138) and B (142), methyl-1-methoxyindole-3-carboxylate (59) and metabolites bis(4-isothiocyanotobutyl)-disulfide (139), 5-(3-isothiocyanato-propylsulfanyl)-pentylisothiocyanate (136) and 3-(methylsulfinyl)-propylisothiocyanate (135). <p>Two metabolites were also isolated from Brussels sprouts and brown mustard; however, these structures are not yet determined. The metabolites 1,4-dimethoxyindole-3-acetonitrile (137) and 5-(3-isothiocyanato-propylsulfanyl)-pentylisothiocyanate (136) are reported for the first time.

The focus of my research was on the secondary metabolites produced by crucifer plants under stress and their biological activity against fungi. Both cultivated and wild plants were investigated to isolate phytoalexins and phytoanticipins, and determine their metabolite profiles.<p>The first chapter of this thesis describes cruciferous plants and their most important pathogenic fungi. These plants are divided into three groups: oilseeds, vegetables and wild species. The metabolites isolated from these plants and their biosynthetic studies are reviewed. In addition economically important necrotrophic fungi such as <i>Leptosphaeria maculans</i>, <i>Alternaria brassicae</i>, <i>Sclerotinia sclerotiorum</i> and <i>Rhizoctonia solani</i> are also reviewed along with their phytotoxins. <p>The second chapter of this thesis describes the detection, isolation, structure determination, syntheses of stress metabolites and biological activity of these metabolites against <i>L. maculans</i>, <i>S. sclerotiorum</i> and <i>R. solani</i>. The investigation of cauliflower led to the isolation of seven phytoalexins: 1-methoxybrassitin (55), spirobrassinin (71), isalexin (64), brassicanal C (60), caulilexins A (106), B (107), and C (105). The phytoalexins caulilexins A (106), B (107) and C (105) were reported for the first time. Caulilexin A (106), having a disulfide bridge, showed the highest activity against S. sclerotiorum and R. solani among the known phytoalexins. Similarly four phytoalexins: 1-methoxybrassitin, brussalexins A (121), B (117) and C (118) along with four metabolites: ascorbigen (51), diindolylmethane (50), 1-methoxy-3,3-diindolylmethane (119) and di-(1-methoxy-3-indolyl)methane (120) were isolated from Brussels sprouts. The phytoalexins brussalexins A (121), B (117) and C (118) are new metabolites. Brussalexin A (121) is the only cruciferous phytoalexins having an allyl thiolcarbamate functional group. The metabolite 1-methoxy-3,3-diindolylmethane (119) is reported for the first time.<p>The investigation of brown mustard for polar metabolites led to the isolation of indole-3-acetonitrile (76) and spirobrassinin (71) along with isorhamnetin-3,7-diglucoside (134). Investigation of wild species such as Asian mustard, sand rocket, wallrocket, hedge mustard and Abyssinian mustard for production of stress metabolites led to the isolation of indole-3-acetonitrile (76), arvelexin (84), 1,4-dimethoxyindole-3-acetonitrile (137), rapalexins A (138) and B (142), methyl-1-methoxyindole-3-carboxylate (59) and metabolites bis(4-isothiocyanotobutyl)-disulfide (139), 5-(3-isothiocyanato-propylsulfanyl)-pentylisothiocyanate (136) and 3-(methylsulfinyl)-propylisothiocyanate (135). <p>Two metabolites were also isolated from Brussels sprouts and brown mustard; however, these structures are not yet determined. The metabolites 1,4-dimethoxyindole-3-acetonitrile (137) and 5-(3-isothiocyanato-propylsulfanyl)-pentylisothiocyanate (136) are reported for the first time.

This thesis was designed to address three seldom studied aspects of the relationship between herbivores and vegetational diversity. 1. Interactions between vegetational diversity and herbivore mortality due to predation were assessed by experimentally manipulating both the species diversity of plants and the densities of a common generalist predator, the spider Enoplagnatha ovata. 2. The importance of plant size to herbivore densities was examined by quantifying plant size (measured as plant height, width, leaf area and growth rates) and adjusting for it through covariate analysis. 3. Differences in population responses of several species of herbivores to both vegetational diversity and a predator, were compared by concurrently studying four lepidopterans. The main experiment used a two factor design, with two planting treatments and two predator treatments. The planting treatments consisted of plots planted with monocultures of brussels sprouts (Brassica oleraceae) and dicultures of brussels sprouts intercropped with peppermint (Mentha piperita). The natural enemy treatments involved augmentations of E ovata in some plots and untreated controls. Two of the herbivores studied, Plutella xylostella and Pieris rapae are monophagous lepidopterans, specializing on crucifers, while the others, Autographa californica and Mamestra configurata are polyphagous. For two species, P. xylostella and M. configurata responses to augmentations of the spider, E. ovata. were different in monocultures and dicultures. Reduced densities of these two species were found in monoculture plots with added spiders; in dicultures increased densities were found in plots with added spiders. This interaction effect points out that generalist predators can be effective in monocultures. I suggest that the importance of natural enemies in monocultures is often overlooked because only the initial colonization phase is being studied. By augmenting predator populations I was able to simulate densities equivalent to those in more established cropping systems. The increased herbivore densities in dicultures with added spiders might be explained by possible predation by E. ovata on other natural enemies of P. xylostella and M. configurata in dicultures but not in monocultures. Supporting evidence for this interpretation lies in the fact that percent parasitism of P. xylostella by the ichneumonid, Diadegma insulare was lower in plots with added spiders than in control plots. Furthermore, parasitism of P. xylostella by D. insulare increased with host density in diculture plots, but not in monoculture plots. Mamestra configurata was not subject to parasitism in this study, precluding assessment of a similar relationship. No A. californica larvae were found in plots with additional spiders. In contrast, P. rapae larvae were not affected by the experimental treatments. Plant size was a crucial determinant of both herbivore populations and percent parasitism of those herbivores. Most importantly, had plant size not been accounted for, the importance of vegetational diversity to both herbivore densities and percent parasitism would have been overestimated. For example, the incorrect conclusion, that vegetational diversity alone was important in determining the abundance of both of the generalist feeders would have been reached. The greater densities of A. californica in monocultures and M. configurata in dicultures were accounted for by plant size. Without plant size adjustments, percent parasitism of P. xylostella by D. insulare would have been misinterpeted as being greater in monocultures than dicultures. With plant size adjustments, the importance of E. ovata augmentations on lowering percent parasitismwas unmasked. All important interaction effects were discovered only after adjustments for plant size had been made. Despite the low densities of all herbivore species, significant responses to experimental treatments were found in three of the four species studied. Only P.l rapae was unaffected by any of the treatments. However, conclusions based on the feeding ng habits of the herbivores could not be made. The polyphagous feeders were affected by generalist predation as much as the crucifer specialist. Parasitism was found in only two of the species, P. xylostella and A. californica. Of these two species parasitism of the specialist, P. xylostella was affected by both vegetational diversity and generalist predation, whereas parasitism of A. californica was not. My study emphasizes multifaceted interactions between the size and diversity of a primary resource and several trophic levels of consumers. Multifactor models, involving several aspects of a cropping system, are required to uncover the important mechanisms behind variable herbivore responses to vegetational diversity. / Land and Food Systems, Faculty of / Graduate

Thesis (M.S.)--University of Delaware, 2006. / Principal faculty advisor: Wallace G. Pill, Dept. of Plant & Soil Sciences. Includes bibliographical references.

Inflammatory diseases such as hypertension are associated with high levels of oxidative stress. Characteristic of oxidative stress is the inflammatory acute phase protein response. Oxidative stress and its accompanied inflammation can be reduced via phase 2 enzyme induction. Broccoli sprouts, a rich source of phase 2 enzyme inducers such as isothiocyanates, can be incorporated into the diet to increase phase 2 enzymes. <p> The hypothesis of this study is that, the dietary intake of dried broccoli sprouts, by inducing liver phase 2 enzymes, will decrease oxidative stress and the acute phase response in the blood of spontaneously hypertensive stroke-prone rats. <p> Spontaneously hypertensive stroke-prone rats (SHRsp) and Sprague Dawley (SD) rats were placed either on a control diet of modified AIN-93G or an experimental diet of modified AIN-93G supplemented with dried broccoli sprouts. The following parameters were examined: 1. Isothiocyanate absorption (an increased level of dithiocarbamates is reflective of ITC absorption), 2. Oxidative stress (a reduction in oxidative stress is evidenced by an increase in plasma protein thiols and blood glutathione (GSH)), 3. Acute phase proteins (a decreased APR is reflected by an increase in plasma albumin and a decrease in ceruloplasmin), 4. Activity of phase 2 enzymes (increased phase 2 enzyme induction results in higher activities of liver quinone reductase (QR), glutathione-S-transferase (GST) and glutathione reductase (GR)). <p> My experimental results demonstrated that broccoli sprout feeding results in higher protein thiol levels in female SHRsp and higher blood GSH levels in males but no acute phase protein changes were observed in either male or female SHRsp. Broccoli sprout feeding caused higher QR and lower GST activities in female SHRsp but did not affect the activities of phase 2 enzymes in male SHRsp. The activities of GST and QR were higher in SD rats than in SHRsp. Levels of dithiocarbamates were higher in the broccoli fed group than in the control fed group. <p> The results from this study do not present a clear pattern to support the hypothesis that dietary intake of broccoli sprouts by inducing phase 2 enzymes will decrease parameters of oxidative stress and the acute phase response. <p> In conclusion, there is an interactive role played by animal gender and the induction of phase 2 enzymes by dried broccoli sprouts.

Inflammatory diseases such as hypertension are associated with high levels of oxidative stress. Characteristic of oxidative stress is the inflammatory acute phase protein response. Oxidative stress and its accompanied inflammation can be reduced via phase 2 enzyme induction. Broccoli sprouts, a rich source of phase 2 enzyme inducers such as isothiocyanates, can be incorporated into the diet to increase phase 2 enzymes. <p> The hypothesis of this study is that, the dietary intake of dried broccoli sprouts, by inducing liver phase 2 enzymes, will decrease oxidative stress and the acute phase response in the blood of spontaneously hypertensive stroke-prone rats. <p> Spontaneously hypertensive stroke-prone rats (SHRsp) and Sprague Dawley (SD) rats were placed either on a control diet of modified AIN-93G or an experimental diet of modified AIN-93G supplemented with dried broccoli sprouts. The following parameters were examined: 1. Isothiocyanate absorption (an increased level of dithiocarbamates is reflective of ITC absorption), 2. Oxidative stress (a reduction in oxidative stress is evidenced by an increase in plasma protein thiols and blood glutathione (GSH)), 3. Acute phase proteins (a decreased APR is reflected by an increase in plasma albumin and a decrease in ceruloplasmin), 4. Activity of phase 2 enzymes (increased phase 2 enzyme induction results in higher activities of liver quinone reductase (QR), glutathione-S-transferase (GST) and glutathione reductase (GR)). <p> My experimental results demonstrated that broccoli sprout feeding results in higher protein thiol levels in female SHRsp and higher blood GSH levels in males but no acute phase protein changes were observed in either male or female SHRsp. Broccoli sprout feeding caused higher QR and lower GST activities in female SHRsp but did not affect the activities of phase 2 enzymes in male SHRsp. The activities of GST and QR were higher in SD rats than in SHRsp. Levels of dithiocarbamates were higher in the broccoli fed group than in the control fed group. <p> The results from this study do not present a clear pattern to support the hypothesis that dietary intake of broccoli sprouts by inducing phase 2 enzymes will decrease parameters of oxidative stress and the acute phase response. <p> In conclusion, there is an interactive role played by animal gender and the induction of phase 2 enzymes by dried broccoli sprouts.