Development of a bacteriophage-based biopesticide for fire blight

Fire blight is an economically important disease of apples and pears that is caused by the bacterium Erwinia amylovora. Control of the disease depends on limiting primaly blosson1 infection in the spring, and rapidly removing infected tissue. The possibility of using phages to control E.amylovora populations has been suggested, but previous studies have. failed to show high treatment efficacies. This work describes the development of a phage-based biopesticide that controls E. amylovora populations under field conditions, and significantly reduces the incidence of fire blight. This work reports the first use ofPantoea agglomerans, a non-pathogenic relative ofE. amylovora, as a carrier for E. amylovora.phages. Its role is to support a replicating population of these phages on blossom surfaces during the period when the flowers are most susceptible to infection. Seven phages and one carrier isolate were selected for field trials from existing collections of 56 E. amylovora phages and 249 epiphytic orchard bacteria. Selection of the . /' phages and carrier was based on characteristics relevant to the production and field perfonnance of a biopesticide: host range, genetic diversity, growth under the conditions of large-scale production, and the ability to prevent E. amylovora from infecting pear blossoms. In planta assays showed that both the phages and the carrier make significant contributions to reducirig the development of fire blight symptoms in pear blossoms. Field-scale phage production and purification methods were developed based on the growth characteristics of the phages and bacteria in liquid culture, and on the survival of phages in various liquid media. Six of twelve phage-carrier biopesticide treatments caused statistically signiflcant reductions in disease incidence during orchard trials. Multiplex real-time PCR was used to simultaneously monitor the phage, carrier, and pathogen populations over the course of selected treatments. In all cases. the observed population dynamics of the biocontrol agents and the pathogen were consistent with the success or failure of each treatment to control disease incidence. In treatments exhibiting a significantly reduced incidel1ce of fire blight, the average blossom population ofE.amylovora had been reduced to pre-experiment epiphytic levels. In successful treatments the phages grew on the P. agglomerans carrier for 2 to 3 d after treatment application. The phages then grew preferentially on the pathogen, once it was introduced into this blossom ecosystem. The efficacy of the successful phage-based treatnlents was statistically similar to that of streptomycin, which is the most effective bactericide currently available for fire blight prevention. The in planta behaviour ofE. amylovora was compared to that ofErwinia pyrifoliae, a closely related species that causes fire blight-like synlptoms on pears in southeast Asia. Duplex real-time PCR was used to monitor the population dynamics of both species on single blossonls. E. amylovora exhibited a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae. The genome ofErwinia phage Ea21-4 was sequenced and annotated. Most of the 8-4.7 kB genome is substantially different from previously described sequences, though some regions are notably similar to Salmonella phage Felix 01 . Putative functions were assigned to approximately 30% of the predicted open reading frames based on amino acid sequence comparisons and N-terminal sequencing of structural proteins.

Structural studies in the Southern Province, South of Sudbury, Ontario

Rocks correlated with the Hough Lake and Quirke Lake Groups of the Huronian Supergroup form part of a northeasterly trending corridor that separates 1750 Ma granitic intrusive rocks of the Chief Lake batholith from the 1850 Ma mafic intrusive rocks of the Sudbury Igneous Complex. This corridor is dissected by two major structural features; the Murray Fault Zone (MFZ) and the Long Lake Fault (LLF). Detailed structural mapping and microstructural analysis indicates that the LLF, which has juxtaposed Huronian rocks of different deformation style and metamorphism grade, was a more significant plane of dislocation than the MFZ. The sense of displacement along the LLF is high angle reverse in which rocks to the southeast have been raised relative to those in the northwest. South of the LLF Huronian rocks underwent ductile defonnation at amphibolite facies conditions. The strain was constrictional, defined by a triaxial strain ellipsoid in which X > Y > z. Calculations of a regional k value were approximately 1.3. Penetrative ductile defonnation resulted in the development of a preferred crystallographic orientation in quartz as well as the elongation of quartz grains to fonn a regional southeast-northwest trending, subvertical lineation. Similar lithologies north of the LLF underwent dominantly brittle deformation under greenschist facies conditions. Deformation north of the LLF is characterized by the thrusting of structural blocks to form angular discordances in bedding orientation which were previously interpreted as folds. Ductile deformation occurred between 1750 and 1238 Ma and is correlated with a regional period of south over north reverse faulting that effected much of the southern Sudbury region. Post dating the reverse faulting event was a period of sedimentation as a conglomerate unit was deposited on vertically bedded Huronian rocks. Rocks in the study area were intruded by both mafic and felsic dykes. The 1238 Ma mafic dykes appear to have been offset during a period of dextral strike slip displacement along the major fault'). Indirect evidence indicates that this event occurred after the thrusting at 950 to 1100 Ma associated with the Grenvillian Orogeny.

Structural analysis of the Paint Lake Deformation Zone, Northern Ontario /

The Paint Lake Deformation Zone (PLDZ), located within the Superior Province of Canada, demarcates a major structural and lithological break between the Onaman-Tashota Terrane to the north and the Beardmore-Geraldton Belt to the south. The PLDZ is an east-west trending lineament, approximately 50 km in length and up to 1 km in width, comprised of an early ductile component termed the Paint Lake Shear Zone and a late brittle component known as the Paint Lake Fault. Structures associated with PLDZ development including S-, C- and C'-fabrics, stretching lineations, slickensides, C-C' intersection lineations, Z-folds and kinkbands indicate that simple shear deformation dominated during a NW-SE compressional event. Movement along the PLDZ was in a dextral sense consisting of an early differential motion with southside- down and a later strike-slip motion. Although the locus of the PLDZ may in part be lithologically controlled, mylonitization which accompanied shear zone development is not dependent on the lithological type. Conglomerate, intermediate and mafic volcanic units exhibit similar mesoscopic and microscopic structures where transected by the PLDZ. Field mapping, supported by thin section analysis, defines five strain domains increasing in intensity of deformation from shear zone boundary to centre. A change in the dominant microstructural deformation mechanism from dislocation creep to diffusion creep is observed with increasing strain during mylonitization. C'-fabric development is temporally associated with this change. A decrease in the angular relationship between C- and C'-fabrics is observed upon attaining maximum strain intensity. Strain profiling of the PLDZ demonstrates the presence of an outer primary strain gradient which exhibits a simple profile and an inner secondary strain gradient which exhibits a more complex profile. Regionally metamorphosed lithologies of lower greenschist facies outside the PLDZ were subjected to retrograde metamorphism during deformation within the PLDZ.

Strategies for economic development in the Commonwealth Caribbean: a comparative study of the foreign economic policy strategies adopted by the Manley government in Jamaica and the Williams government in Trinidad and Tobago

This thesis compares the foreign economic poUcy dimension of the development strategies adopted by the governments of two Commonwealth caribbean countries: The Hardey government In Jamaica, and the· Williams government in Trlnidad and T ooago, The foreign economic policIes adopted by these governments appeared, on the surface~ to be markedly dissimilar. The Jamakan strategv on the one hand, emphasised self-reliance and national autonomy; and featured the espousal of radical oonaHgnment together with attempts to re-deftne the terms of the Islands externaa economIc relaUoos. The Trinidadian strategy 00 the other hand, featured Uberal externaUy-oriented growth poUctes, and close relatjoos with Western governments and financial institutions. Th1s study attempts to identify the explanatory factors that account for the apparent dlssimUarUy 1n the foreign economic policies of these two govemnents. The study is based on a comparison of how the structural bases of an underdeveloped ecooomYg and the foreign penetration and vulnerabUUy to external pressures asSOCiated wUh dependence, shape and influence foreign economic poUcy strategy. The framework views fore1gn ecooom1c strategy as an adaptive response on the part of the decision makers of a state to the coostralnts and opportunities provided by a particular situation. The · situat i 00' in this case being the events, conditions, structures and processes, associated wUh dependente and underdevelopment. The results indicate that the similarities and dissimHarities in the foreign economic policies of the governments of Jamaica and Trinidad were a reflecUon of the simHarities and dissimilarities in their respective situations. The conclusion derived suggests that If the foreign pol1cy field as an arena of choice, Is indeed one of opportunities and constraints for each and every state, then poHcy makers of smaU, weak, hlghW penetrated and vulnerable states enter thlS arena with constraints outweighing opportunities. This places effective limits 00 their decisional latitude and the range of policy options avaUable. Policy makers thus have to decide critical issues with few estabUshed precedents, in the face of domestic social and political cleavages, as wen as serious foreign pressures. This is a reflection not only of the trappings of dependence, but also of the Umned capabilities arising from the sman size of the state, and the Impact of the resource-gap In an underdeveloped economy. The Trinidadian strategy 1s UlustraUve of a development strategy made viable through a combination of a fortuitous circumstance, a confluence of the interests of influential groups» and accurate perception on the part of poUcy makers. These factors enabled policy makers to minimise some of the constraints of dependence. The faUure of Manlets strategy on the other hand, 15 iHustraUve of the problems involved tn the adoption of poUcles that work against the interest of internal and external political and economic forces. It is also tUustraUve of the consequences of the faUure 00 the part of policy makers to clarify goals, and to reconcile the values of rapid economic growth with increased self-reliance and national autonomy. These values tend to be mutuany Incompatible given the existing patterns of relations in the jnternational economy.

The effects of computer assisted interactive fiction on the development of reading comprehension in grade five students

This study was undertaken in order to determine the effects of playing computer based text adventure games on the reading comprehension gains of students. Forty-five grade five students from one elementary school were randomly assigned to experimental and control groups, and were tested with regard to ability, achievement and reading skills. An experimental treatment, consisting of playing computer based interactive fiction games of the student's choice for fifteen minutes each day over an eight-week period, was administered. A comparison treatment engaged the control group in sustained silent reading of materials of the student's choice for an equal period of time. Following the experimental period all students were post-tested with an alternate form of the pre-test in reading skills, and gain scores were analysed. It was found that there were no significant differences in the gain scores of the experimental and control groups for overall reading comprehenSion, but the experimental group showed greater gains than the control group in the structural analysis reading sub-skill. Extreme variance in the data made generalization very difficult, but the findings indicated a potential for computer based interactive fiction as a useful tool for developing reading sl

Iridium(I) complexes of phenanthroline-derived benzimidazolylidenes : synthetic, structural and catalytic studies

N-heterocyclic carbenes (NHCs) have undergone rapid development in recent years. Due to their strong a-electron donation and structural variability properties, NHCs are becoming a major class of ligands in organometallic chemistry. Compared with the other two types of NHCs (imidazolylidenes and imidazolinylidenes), benzimidazolylidenes have not been well represented. Limited synthetic approaches may impede the development ofbenzimidazolylidenes. This thesis is focused on the synthesis of phenanthroline-derived benzimidazolylidene ligands and their metal complexes. A series of benzimidazolylidene-iridium complexes were synthesized and characterized spectroscopically and crystallographic ally. All of the new complexes showed varying degrees of catalytic activity and enantioselectivity toward transfer hydrogenation and asymmetric hydrogenation. The best results were achieved in hydrogenation of methyl-2-acetamidoacrylate, which afforded (-)-(R)-methyl-2-acetamidopropanoate in 97% yield and 81 % ee.

Effect of Polymer Architecture on the Structural and Biophysical Properties of PEG-PLA Nanoparticles

Polymers made of poly(ethylene glycol) chains grafted to poly(lactic acid) chains (PEG-g-PLA) were used to produce stealth drug nanocarriers. A library of comb-like PEG-g-PLA polymers with different PEG grafting densities was prepared in order to obtain nanocarriers with dense PEG brushes at their surface, stability in suspension, and resistance to protein adsorption. The structural properties of nanoparticles (NPs) produced from these polymers by a surfactant-free method were assessed by DLS, zeta potential, and TEM and were found to be controlled by the amount of PEG present in the polymers. A critical transition from a solid NP structure to a soft particle with either a “micelle-like” or “polymer nano-aggregate” structure was observed when the PEG content was between 15 to 25% w/w. This structural transition was found to have a profound impact on the size of the NPs, their surface charge, their stability in suspension in presence of salts as well as on the binding of proteins to the surface of the NPs. The arrangement of the PEG-g-PLA chains at the surface of the NPs was investigated by 1H NMR and X-ray photoelectron spectroscopy (XPS). NMR results confirmed that the PEG chains were mostly segregated at the NP surface. Moreover, XPS and quantitative NMR allowed quantifying the PEG chain coverage density at the surface of the solid NPs. Concordance of the results between the two methods was found to be remarkable. Physical-chemical properties of the NPs such as resistance to aggregation in saline environment as well as anti-fouling efficacy were related to the PEG surface density and ultimately to polymer architecture. Resistance to protein adsorption was assessed by isothermal titration calorimetry (ITC) using lysozyme. The results indicate a correlation between PEG surface coverage and level of protein interactions. The results obtained lead us to propose such PEG-g-PLA polymers for nanomedecine development as an alternative to the predominant polyester-PEG diblock polymers.

Synthesis, Spectral Characterization, Structural Studies and Biological Investigations of Transition Metal Complexes of Some Acid Hydrazones

This study concentrates the chemical properties of hydrazones due to its chelating capability and their pharmacological applications. Studies cover the preparation of different acid hydrazones and their structural studies and studies on their antimicrobial activity, synthesis and spectral characterization of different complexes of copper oxovanadium, manganese, nickel etc. Effect of incorporation of heterocyclic bases to the coordination sphere, change in the biological activity of acid hydrazones upon coordination, development of X-ray quality single crystals and its X-ray diffraction studies, studies on the redox behavior of the coordinated metal ions and correlation between the stereochemistry and biological activities.

Structural and Spectral Investigations of Transition Metal Complexes Of Di-2-Pyridyl Ketone N (4), N (4)-Disubstituted Thiosemicarbazones

The study deals with structural and spectral investigations of transition metal complexes of di-2-pyridyl ketone N(4),N(4)-disubstituted thiosemicarbazones. The main objective and scope of the work deals with di-2-pyridyl ketone N(4),N(4)-disubstituted thiosemicarbazones are quardridentate NNNS donor ligands. To chosen this ligand for study because, the ligands are prepared and characterized for the first time, since there are two pyridyl nitorgens, dimmers and polymers of complexes may result leading to interesting structural aspects. The work includes the preparation of the thiosemicarbzones and their structural and spectral studies, synthesis and spectral characterization of complexes of copper(II),,nickel(II),manganese(II), dioxovanadium(V),cobalt(III),zinc(II),cadmium(II) of the ligand HL, synthesis and spectral characterization of complexes of copper(II),manganese(II), of the ligand HL and the development of X-ray quality crystals and its X-ray diffraction studies. The structural characterization techniques are elemental analysis, conductivity measurements, magnetic measurements, electronic spectroscopy, H NMR spectroscopy, Infrared spectroscopy and X-ray crystallography.

A STUDY ON SOFTWARE DEVELOPMENT PROJECT RISK, RISK MANAGEMENT, PROJECT OUTCOMES AND THEIR INTER-RELATIONSHIP

This research was undertaken with an objective of studying software development project risk, risk management, project outcomes and their inter-relationship in the Indian context. Validated instruments were used to measure risk, risk management and project outcome in software development projects undertaken in India. A second order factor model was developed for risk with five first order factors. Risk management was also identified as a second order construct with four first order factors. These structures were validated using confirmatory factor analysis. Variation in risk across categories of select organization / project characteristics was studied through a series of one way ANOVA tests. Regression model was developed for each of the risk factors by linking it to risk management factors and project /organization characteristics. Similarly regression models were developed for the project outcome measures linking them to risk factors. Integrated models linking risk factors, risk management factors and project outcome measures were tested through structural equation modeling. Quality of the software developed was seen to have a positive relationship with risk management and negative relationship with risk. The other outcome variables, namely time overrun and cost over run, had strong positive relationship with risk. Risk management did not have direct effect on overrun variables. Risk was seen to be acting as an intervening variable between risk management and overrun variables.

“Design and Development of Coplanar Strip Fed Planar Antennas”

With the recent progress and rapid increase in the field of communication, the designs of antennas for small mobile terminals with enhanced radiation characteristics are acquiring great importance. Compactness, efficiency, high data rate capacity etc. are the major criteria for the new generation antennas. The challenging task of the microwave scientists and engineers is to design a compact printed radiating structure having broadband behavior along with good efficiency and enhanced gain. Printed antenna technology has received popularity among antenna scientists after the introduction of planar transmission lines in mid-seventies. When we view the antenna through a transmission line concept, the mechanism behind any electromagnetic radiator is quite simple and interesting. Any electromagnetic system with a discontinuity is radiating electromagnetic energy. The size, shape and orientation of the discontinuities control the radiation characteristics of the system such as radiation pattern, gain, polarization etc. It can be either resonant or non-resonant. This thesis deals with antennas that are developed from a class of transmission lines known as coplanar strip-CPS, a planar analogy of parallel pair transmission line. The specialty of CPS is its symmetric structure compared to other transmission lines, which makes the antenna structures developed from CPS quite simple for design and fabrication. The structural modifications on either metallic strip of CPS results in different antennas. The first part of the thesis discusses a single band and dual band design derived from open ended slot lines which are very much suitable for 2.4 and 5.2 GHz WLAN applications. The second section of the study is vectored into the development of enhanced gain dipoles. A single band dipole and a wide band enhanced gain dipole suitable for 5.2/5.8 GHZ band and imaging applications are developed and discussed. Last part of the thesis discusses the development of directional UWBs. Three different types of ultra-compact UWBs are developed and almost all the frequency domain and time domain analysis of the structures are discussed.

Structural characterization and distribution of zebrafish germ plasm during interphase and mitosis

In zebrafish, germ cells are responsible for transmitting the genetic information from one generation to the next. During the first cleavages of zebrafish embryonic development, a specialized part of the cytoplasm known as germ plasm, is responsible of committing four blastomeres to become the progenitors of all germ cells in the forming embryo. Much is known about how the germ plasm is spatially distributed in early stages of primordial germ cell development, a process described to be dependant on microtubules and actin. However, little is known about how the material is inherited after it reorganizes into a perinuclear location, or how is the symmetrical distribution regulated in order to ensure proper inheritance of the material by both daughter cells. It is also not clear whether there is a controlled mechanism that regulates the number of granules inherited by the daughter cells, or whether it is a random process. We describe the distribution of germ plasm material from 4hpf to 24hpf in zebrafish primordial germ cells using Vasa protein as marker. Vasa positive material appears to be conglomerate into 3 to 4 big spherical structures at 4hpf. While development progresses, these big structures become smaller perinuclear granules that reach a total number of approximately 30 at 24hpf. We investigated how this transformation occurs and how the minus-end microtubule dependent motor protein Dynein plays a role in this process. Additionally, we describe specific colocalization of microtubules and perinuclear granules during interphase and more interestingly, during all different stages of cell division. We show that distribution of granules follow what seems to be a regulated distribution: during cells division, daughter cells inherit an equal number of granules. We propose that due to the permanent colocalization of microtubular structures with germinal granules during interphase and cell division, a coordinated mechanism between these structures may ensure proper distribution of the material among daughter cells. Furthermore, we show that exposure to the microtubule-depolymerizing drug nocodazole leads to disassembly of the germ cell nuclear lamin matrix, chromatin condensation, and fusion of granules to a big conglomerate, revealing dependence of granular distribution on microtubules and proper nuclear structure.

Parameter identification of structural dynamic models by inverse statistical analysis

Auf dem Gebiet der Strukturdynamik sind computergestützte Modellvalidierungstechniken inzwischen weit verbreitet. Dabei werden experimentelle Modaldaten, um ein numerisches Modell für weitere Analysen zu korrigieren. Gleichwohl repräsentiert das validierte Modell nur das dynamische Verhalten der getesteten Struktur. In der Realität gibt es wiederum viele Faktoren, die zwangsläufig zu variierenden Ergebnissen von Modaltests führen werden: Sich verändernde Umgebungsbedingungen während eines Tests, leicht unterschiedliche Testaufbauten, ein Test an einer nominell gleichen aber anderen Struktur (z.B. aus der Serienfertigung), etc. Damit eine stochastische Simulation durchgeführt werden kann, muss eine Reihe von Annahmen für die verwendeten Zufallsvariablengetroffen werden. Folglich bedarf es einer inversen Methode, die es ermöglicht ein stochastisches Modell aus experimentellen Modaldaten zu identifizieren. Die Arbeit beschreibt die Entwicklung eines parameter-basierten Ansatzes, um stochastische Simulationsmodelle auf dem Gebiet der Strukturdynamik zu identifizieren. Die entwickelte Methode beruht auf Sensitivitäten erster Ordnung, mit denen Parametermittelwerte und Kovarianzen des numerischen Modells aus stochastischen experimentellen Modaldaten bestimmt werden können.

Family agriculture for bottom-up rural development: a case study of the indigenous Mayan population in the Mexican Peninsula

Since pre-colonial times the indigenous communities of Mayan origin in the state of Quintana Roo, Mexico, widely practice home gardens on a sustainable basis as the principal form of family agriculture. This study analyzes the structural complexity, functional diversity and management strategy of these indigenous home gardens in order to attempt to propose recommendations for improved family farming. The Mayan home gardens are structured into three or more vertical layers of multiple plant species of herbs, shrubs and trees, and horizontally into well-defined zones for production of both domestic and wild animals. The home gardens provide multiple services apart from food and nutrition security. For sustainable bottom-up rural development, we recommend the continuation of multifunctional home gardens.

Design and Development of 3-DOF Modular Micro Parallel Kinematic Manipulator

This paper presents the research and development of a 3-legged micro Parallel Kinematic Manipulator (PKM) for positioning in micro-machining and assembly operations. The structural characteristics associated with parallel manipulators are evaluated and the PKMs with translational and rotational movements are identified. Based on these identifications, a hybrid 3-UPU (Universal Joint-Prismatic Joint-Universal Joint) parallel manipulator is designed and fabricated. The principles of the operation and modeling of this micro PKM is largely similar to a normal size Stewart Platform (SP). A modular design methodology is introduced for the construction of this micro PKM. Calibration results of this hybrid 3-UPU PKM are discussed in this paper.