Thesis (M.S.)--Ball State University, 2009. / Title from PDF t.p. (viewed on May 07, 2010). Includes bibliographical references (p. 30-38).

Thesis (M.A.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / Serotonin (5-hydroxytryptamine), one of the biologically active amines, has been found in rather large quantities in the dorsal skin of Rana pipiens. Quantitative determination of the endogenous levels of the amine in the whole dorsal skin homogenate extracts were made on a Farrand spectroflourometer, and ranges of 120-200 micrograms per gram of fresh tissue were established. Comparison of whole skin homogenates and homogenates of just the dorsal cutaneous ridges, or dermal plicae, indicated that the amine is localized in these ridges. Acid treatment of the denser more fibrous portions of the homogenate prior to extraction more then noublcc the yields of serotonin. This result showed that the amine is found primarily in the deeper portions of the skin, very likely in the poison glands which are concentrated in the dermal plicae. Skin homogenates were also studied to determine if serotonin is actually synthesized in the skin. A precursor of the amine, 5-hydroxytryptophan, decarboxylates in tne presence of 5-hydroxytryrtophan decarooxylase to produce the amine. 5-hydroxytryrtophan was incubated with the homogenate, under nitrogen, for two hours to determine enzymic activity. Under the conditions of the experiment, no enzymic activity could be shown, indicating that serotonin is not produced in the skin of the frog in detectable amounts. The fact that serotonin levels are so high, and the lack of enzymic activity indicates that once synthesized serotonin is stored in an inactive form or is synthesized elsewhere and transnorted to the skin. / 2031-01-01

No description available.

Culex pipiens, in the family Culicidae, has emerged as one of the biggest vectors for West Nile virus. It has two bioforms, pipiens and molestus, which differ from each other regarding habitat, diapause, and prey. Pipiens prefers to bite birds, and molestus prefers to bite humans. There is to some extent hybridization between the two, which creates a bridge-vector between birds and humans. One way to study the relationships and spreading of the mosquitos is using the intracellular bacteria Wolbachia pipientis which is present in at least 99% of al Culex mosquitoes. The bacteria have two fast evolving genes, pk1 and ank2 which are suitable for this. Not only are the bacteria suitable for genetics, but it is also manipulating the reproductive system of the mosquitoes through Cytoplasmic Incompatibility, which changes structures of populations and allows for the bacteria to spread fast and efficient. We wanted to investigate levels of Wolbachia in different populations, as well as if the two bioforms prefer a prey, together with mapping the relationships between populations using the two genes. We found that Wolbachia was present in all tested mosquitoes, with higher levels of it in the abdomen than in the thorax. We also found that the theory of a preferred prey was false within the tested populations, since both bioforms preferred birds. Lastly, we could identify five different strains of Wolbachia pipientis concentrated to different locations. This study has shown that Wolbachia is present in all tested mosquitoes and is a useful tool to determine relationships within and between populations. This is important to be able to gain understanding of the spread of West Nile virus and other vector borne diseases spread by Culex pipiens mosquitoes.

Access to abstract permanently restricted to Ball State community only / Access to thesis permanently restricted to Ball State community only / Department of Biology

This study was undertaken to determine the effects of beryllium nitrate on regeneration in the tail of Rana Pipiens larvae. The tails were amputated approximately 3 rom from the tips. Some were then treated with different concentrations of beryllium nitrate (0.5 N, 0.7 N, and 1.0 N). Normal regeneration of untreated amputated tails was fulfilled in an orderly manner. In the animals treated with beryllium nitrate, general tissue regeneration was inhibited. Considerable degeneration was observed in all tissues of the tail stumps. Holfever, the elastic lamella of the notochordal sheath was stimulated to stretch, oftentimes breaking the continuity of the fibrous lamella. The elastic layer appeared highly ravelled within the degenerated tissues of the tail, and, in some cases, extended for 3 to 4 inches from the tail stump due to a break in the covering epithelium.

Host-pathogen investigations have conceptually evolved during the last two decades, from a basic and descriptive approach to a current hypothesis-driven and a more theoretical discipline shaped by evolutionary biology. Our deeper understanding of the elements influencing the mutual selective pressures that the host and the pathogens exert on each other, together with recent conceptual advances, currently position this field of research at the frontier between ecology and evolution. Recent theoretical considerations define hostpathogens systems as an evo-eco mosaic comprised of evolutionary and ecological attributes in turn underlying the context-dependent nature of the system dynamic. Therefore, investigations of host-pathogen interactions should integrate the diversity of the systems drivers by using an integrative approach in order to elucidate both coevolutionary trajectory and epidemiological dynamic of the system. In this thesis, such a framework is used to investigate Amphibian/ranavirus interactions. Ranaviruses are emerging pathogens known to have caused amphibian die-offs on five continents with the greatest number of reported mortality events documented in North America and Europe. Despite an increasing understanding of ranaviral disease properties, ranavirus disease dynamics in the environment remain poorly understood. For instance, the influence of potential abiotic and biotic mechanisms including temperature, local landscape features, larval developmental stages, host density and genetic variability as well as genotypic interactions between the host and the pathogen has on the prevalence and virulence of the virus remains to be elucidated. In order to improve our knowledge regarding these specific determinants of ranaviral disease, I designed a combination of manipulative laboratory experiments and a field mensurative survey using the ranid amphibian Lithobates (Rana) pipiens as the host model for this system. I observed that populations of amphibian hosts inhabiting urbanized landscapes suffered from significant decline in genetic diversity in turn promoting the accrued infection by the ranavirus (manuscript 1). Complementary analysis using two amphibian host species, L.pipiens and L.sylvaticus, and three ranavirus strains revealed significant variation among hosts for their susceptibility to ranavirus, and significant variation among ranavirus strains for infectivity. I also showed that specific amphibian/ranavirus interactions might have a tighter coevolutionary history than other combinations, resulting in sharper mutual coadaptations and the potential for frequency-dependent selection to operate in this system. However, the coevolutionary trajectories in this host-pathogen system are dependent on the temperature conditions in which the interaction takes place. Amphibian/ranavirus interactions outcomes iv are therefore temperature, host, and pathogen genotype-dependent suggesting that the range of infection outcomes in this system is potentially large (manuscript 2). Further, I observed that increasing animal holding density is detrimental for host fitness as mortality rate is higher, day of death earlier, development longer, and growth rate significantly lower when tadpoles are experimentally exposed to ranavirus in high holding density situations. These results paralleled a linear increase of detrimental effects when ranavirus doses increased in low density conditions, with control tadpoles having a significantly higher overall relative fitness. However, this pattern was not observed in high density conditions, where the effects of increasing ranavirus dose were limited, revealing non-trivial density-dependence of virulence expression (manuscript 3). Finally, ranavirus infection rate varied with the host developmental stage as the host immune system clears the infection over the course of individual host development. However the intensity of the clearing depends on both the timing and number of ranavirus exposures (manuscript 4). Overall the results described in my thesis suggest that ranavirus virulence depends on a diversity of ecological, epidemiological, and evolutionary determinants. The underlying complexity of ranavirus

Les Wolbachia sont des alpha-protéobactéries intracellulaires transmises verticalement de la mère aux descendants via les ovocytes. Du fait de ce mode de transmission, des stratégies de manipulation de la reproduction favorisant leur propagation ont été sélectionnées chez ces bactéries. La plus communément utilisée par Wolbachia s’appelle l’incompatibilité cytoplasmique (IC). L’IC a lieu lorsque des mâles infectés copulent avec des femelles non infectées ou infectées par des Wolbachia incompatibles et se traduit par la mort des descendants avant l’éclosion. L’IC est généralement conceptualisée comme un modèle mod/resc ou toxine/antidote dans lequel les Wolbachia présentes chez les mâles introduiraient dans les spermatozoïdes une toxine (fonction mod) qui, après la fécondation, entrainerait la mort des embryons, sauf si les Wolbachia présentes dans l’œuf produisent un antidote (fonction resc). C’est chez l’espèce de moustique Culex pipiens que la plus grande diversité de phénotypes d’IC a été décrite. Cette diversité repose uniquement sur la diversité des souches de Wolbachia hébergées par C. pipiens. Dans cette thèse, nous avons mené la première étude des mécanismes cytologiques responsables de la mort des embryons dans les croisements incompatibles chez C. pipiens. Nous avons montré que des défauts de condensation et de ségrégation de la chromatine paternelle lors de la première division embryonnaire entraînent la mort des embryons dans tous les croisements incompatibles étudiés. Ces défauts cellulaires sont les seuls observés, ce qui indique que la diversité de relations d’IC décrite chez C. pipiens ne repose pas sur une diversité de défauts cellulaires. L’étude, chez plusieurs souches de wPips, de l’opéron cidA/cidB dont l’implication fonctionnelle dans l’IC a récemment été mise en évidence chez la drosophile, nous a permis de montrer que cet opéron est amplifié et polymorphe dans tous les génomes de wPips séquencés. L’exploration des variants de cet opéron dans les génomes de Wolbachia infectant des populations naturelles de C. pipiens, à l’aide de plus de 250 lignées isofemelles, a permis d’associer de manière robuste une variation de cidB avec un changement dans le phénotype mod de certains mâles. En outre, la présence d’un variant ubiquitaire de cidA, supporte le rôle de ce gène dans la fonction resc. Ainsi, chez C. pipiens, l’opéron cidA/cidB grâce à son amplification et sa diversification est impliqué dans la diversité des phénotypes d’IC et fonctionnerait comme un système toxine-mod /antidote-resc : cidB étant impliqué dans la fonction mod et cidA dans la fonction resc. / Wolbachia are intracellular alpha-proteobacteria vertically transmitted from mothers to their offspring through oocytes. As a consequence of this transmission mode, reproductive manipulation strategies that promote bacteria spread in host populations have been selected. The most common manipulation used by Wolbachia is called cytoplasmic incompatibility (CI). CI occurs when infected males mate with uninfected or incompatible Wolbachia-infected females and results in the death of offspring before hatching. CI is generally conceptualized as a mod/resc or toxin/antidote model in which paternal Wolbachia would introduce a toxin (mod function) in sperms which would, after fertilization, induce embryonic death unless an antidote produced by maternal Wolbachia in the egg counteracts its effect (resc function). A to date unique diversity of CI phenotypes has been described in the mosquito species Culex pipiens. This diversity is based solely on the diversity of Wolbachia strains hosted by C. pipiens. In this PhD, we conducted, in C. pipiens, the first study of the cytological mechanism behind embryonic mortality in CI crosses. We showed that paternal chromatin condensation and segregation defects during the first embryonic division were responsible for embryonic death in all CI crosses. These CI defects were the only ones observed indicating that the diversity of CI phenotypes in C. pipiens is not based on a diversity of cellular mechanisms. We then studied the cidA/cidB operon in several wPip strains as the functional involvement of this operon in CI was recently demonstrated in Drosophila. We showed that this operon is amplified and polymorphic in all genomes of sequenced wPip. Investigation of cidA/cidB variants in Wolbachia genomes infecting natural populations of C. pipiens, using more than 250 isofemale lines, enabled us to reveal a robust association between cidB variations and change in mod phenotype. In addition, the presence of an ubiquitous cidA variant supports the role of this gene in the resc function. In C. pipiens, the cidA/cidB operon, through its amplification and diversification, is involved in the CI phenotypes diversity and would operate as a toxin-mod / antidote-resc system: cidB being involved in the mod function and cidA in the resc function.

<p>Meteorological&nbsp; conditions&nbsp; have&nbsp; a&nbsp; significant influence on the time of occurrence, abundance and activity of the mosquito vector. In the current context of climate change, it is of great importance to assess the&nbsp; impact&nbsp; of&nbsp; shifts&nbsp; in&nbsp; climatic&nbsp; conditions&nbsp; on&nbsp; the suitability for the establishment and annual activity of&nbsp; the&nbsp; vector&nbsp; species.&nbsp; Moreover,&nbsp; changes&nbsp; in&nbsp; the variability&nbsp; of&nbsp; meteorological&nbsp; elements&nbsp; and&nbsp; their extremes&nbsp; can&nbsp; generate&nbsp; unexpected&nbsp; changes&nbsp; in&nbsp; the mosquito&nbsp; vector&nbsp; population&nbsp; which&nbsp; in&nbsp; turn&nbsp; have&nbsp; an&nbsp; important effect on human health. One of the ways to put&nbsp; these&nbsp; causes&nbsp; and&nbsp; effects&nbsp; into&nbsp; perspective&nbsp; is&nbsp; to simulate the activity of the vector within a processbased framework which allows for the analysis of the contribution of individual factors on the different life stages of the vector. Such analysis is presented by use of&nbsp; sophisticated&nbsp; dynamical&nbsp; models&nbsp; simulating&nbsp; the characteristics of the biological population, forced by observed&nbsp; meteorological&nbsp; data,&nbsp; capturing&nbsp; the&nbsp; localmicro-environment&nbsp; of&nbsp; the&nbsp; vector&nbsp; habitat,&nbsp; and validated by the observed entomology.Numerical&nbsp; models&nbsp; are&nbsp; being&nbsp; developed&nbsp; to&nbsp; model vector&nbsp; population&nbsp; dynamics&nbsp; and&nbsp; the&nbsp; expected circulation of the virus within a closed system. Two modelling&nbsp; approaches&nbsp; are&nbsp; standardly&nbsp; applied&nbsp; to modelling vector population dynamics: Mechanistic and&nbsp; Stochastic.&nbsp; The&nbsp; advantage&nbsp; of&nbsp; mechanistic&nbsp; over<br />statistical&nbsp; models&nbsp; is&nbsp; that&nbsp; they&nbsp; can&nbsp; provide&nbsp; a deterministic&nbsp; framework&nbsp; allowing&nbsp; for&nbsp; the&nbsp; isolated evaluation of each input parameter and their effect on the modelled system. Mechanistic dynamical models are used to describe the biophysical processes or part<br />of&nbsp; the&nbsp; process&nbsp; as&nbsp; a&nbsp; response&nbsp; to&nbsp; changes&nbsp; in&nbsp; the meteorological conditions.<br />The&nbsp; work&nbsp; carried&nbsp; out&nbsp; in&nbsp; this&nbsp; thesis&nbsp; can&nbsp; be summarized as follows: (i) Analysis of the association between&nbsp; the&nbsp; most&nbsp; important&nbsp; abiotic&nbsp; drivers influencing the population dynamics, annual activity and&nbsp; time&nbsp; of&nbsp; occurrence&nbsp; of Culex&nbsp; pipiens and Aedes aegypti;&nbsp; (ii)&nbsp; Identifying&nbsp; the&nbsp; most&nbsp; important&nbsp; climatic factors and model settings as a function of climatic characteristics of the study region; (iii) Modelling the vector population dynamics and stability analysis of the&nbsp; dynamical&nbsp; system&nbsp; (iv)&nbsp; Analysis&nbsp; of&nbsp; different verification techniques and implications in terms of model&nbsp; application;&nbsp; (v)&nbsp; Feasibility&nbsp; analysis&nbsp; of<br />improving&nbsp; the&nbsp; model&nbsp; with&nbsp; a&nbsp; Land-Surface Parametrization scheme and short-range forecasting of pest population dynamics.</p> / <p>Meteorolo&scaron;ki&nbsp; uslovi&nbsp; bitno&nbsp; utiču&nbsp; na&nbsp; vreme&nbsp; pojave, brojnost&nbsp; vektora&nbsp; i&nbsp; njihovu&nbsp; aktivnost.&nbsp; U&nbsp; uslovima evidentnih promene klime, od ogromne je važnosti sagledati&nbsp; uticaj&nbsp; očekivanih&nbsp; promena&nbsp; klime&nbsp; na pogodnost&nbsp; uslova&nbsp; na&nbsp; pojavu&nbsp; izabranih&nbsp; vektora.<br />Takodje, značajne promene kolebanja meteorolo&scaron;kih elemenata&nbsp; u&nbsp; odnosu&nbsp; na&nbsp; vi&scaron;egodi&scaron;nji&nbsp; prosek&nbsp; i&nbsp; sve če&scaron;će pojave nepovoljnih vremenskih prilika dovode do neočekivanog pona&scaron;anja populacije komarca &scaron;to značajno&nbsp; utiče&nbsp; na&nbsp; kvalitet&nbsp; života&nbsp; i&nbsp; zdravlje&nbsp; ljudi. Jedini&nbsp; način&nbsp; da&nbsp; se&nbsp; sagledaju&nbsp; uzroci&nbsp; i&nbsp; posledice navedenih pojava zasniva se na simulaciji aktivnosti i&nbsp; brojnosti&nbsp; vektora&nbsp; uz&nbsp; mogućnost&nbsp; testiranja&nbsp; uticaja svakog&nbsp; pojedinačnog&nbsp; faktora.&nbsp; Ovu&nbsp; mogućnost pružaju samo visoko sofistikovani dinamički modeli koju su pro&scaron;li proces kalibracije i validacije zasnovan<br />na izmerenim vrednostima meteorolo&scaron;kih elemenata i karakteristika biolo&scaron;ke populacije.<br />Sofistikovani&nbsp; modeli&nbsp; za&nbsp; simulaciju&nbsp; dinamike populacije vektora i očekivane cirkulacije vektorskih transmisivnih bolesti se koriste sa ciljem modeliranja potencijalnog rizika od zaraze i epidemije. Modeli zasimulaciju dinamike vektora mogu da se podele na dve&nbsp; glavne&nbsp; grupe:&nbsp; Mehanističke&nbsp; i&nbsp; Statističke. Prednost&nbsp; mehanističkih&nbsp; modela&nbsp; nad&nbsp; statističkim&nbsp; je &scaron;to&nbsp; mogu&nbsp; da&nbsp; se&nbsp; koriste&nbsp; za&nbsp; evaluaciju&nbsp; uticaja izolovanog&nbsp; faktora&nbsp; na&nbsp; dinamički&nbsp; sistem&nbsp; i odgovarajuće promene brojnosti unutar svake faze u razvoju&nbsp; vektora.&nbsp; Mehanistički&nbsp; dinamički&nbsp; sistemi&nbsp; se koriste&nbsp; kako&nbsp; bi&nbsp; se&nbsp; opisao&nbsp; mehanizam&nbsp; biofizičkog procesa&nbsp; ili&nbsp; dela&nbsp; procesa&nbsp; u&nbsp; zavisnosti&nbsp; od&nbsp; forsirajuće veličine.<br />Predmet&nbsp; istraživanja&nbsp; u&nbsp; ovom&nbsp; radu&nbsp; jeste identifikovanje&nbsp; najznačajnijih&nbsp; biolo&scaron;kih&nbsp; i&nbsp; fizičkih procesa&nbsp; kao&nbsp; i&nbsp; odgovarajućih&nbsp; faktora&nbsp; koji&nbsp; utiču&nbsp; na brojnost i aktivnost vektora roda Aedes i Culex. Ciljevi istraživanja mogu da se sumiraju na sledeći način: (i)<br />analiza najznačajnijih meteorolo&scaron;kih parametara koji utiču na vreme pojave, brojnost i aktivnost vektora Aedes&nbsp; i&nbsp; Culex&nbsp; roda;&nbsp; (ii)&nbsp; definisanje&nbsp; najznačajnijih klimatskih&nbsp; faktora&nbsp; i&nbsp; stepena&nbsp; osetljivosti&nbsp; procesa&nbsp; na njih; (iii) modeliranje dinamike populacije vektora i analiza&nbsp; stabilnosti&nbsp; dinamičkog&nbsp; sistema;&nbsp;&nbsp; (iv) verifikacija&nbsp; i&nbsp; analiza&nbsp; metoda&nbsp; verifikacije&nbsp; i&nbsp; validacije dinamičkog&nbsp; modela;&nbsp; (v)&nbsp; kratkoročna&nbsp; prognoza dinamike&nbsp; populacije&nbsp; komarca&nbsp; i&nbsp; formulacija hidrolo&scaron;kog modula upotrebom SURFEX povr&scaron;inske<br />&scaron;eme sa ECOCLIMAP fiziogeografskim podacima.</p>

Thesis (Ph.D.)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / Quantitative relationships between blood flow velocity, vessel diameter, width of the peripheral plasma layer and induction of thrombus formation were studied in small blood vessels of the retrolingual membrane and intestinal mesentery of the frog, Rana Pipiens, and of the cheek pouch of the golden hamster, Mesocricetus auratus. Blood flow velocity was measured by a modification of the technique described by Hugues (Arch. Int. de Physiol. 61: 565, 1953). Internal vessel diameters and widths of the total peripheral plasma layer were measured with an ocular micrometer. Thrombus thresholds were determined by graded electrical stimulation. Determinations were made on 202 vessels in retrolingual membranes of frogs prepared by single pithing. In 95 arterioles (diameters from 10 to 40 microns), the mean flow velocity was 2.84 mm/sec. and the mean volume flow rate, 1.68xl0-3 cu mm/sec. The mean width of the total peripheral plasma layer was 3.6 microns and its ratio to mean vessel diameter was 1:6. In 107 venules (diameters from 10 to 50 microns), the mean flow rate was 1.20 mm/sec. The mean volume flow rate was 1.02x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 4.1 microns and its ratio to mean vessel diameter was 1:7.4. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.3 and 1:1.6 respectively. Measurements were made on 100 mesenteric vessels of frogs prepared by single pithing. In 50 arterioles (diameters from 15 to 45 microns), the mean velocity was 3.77 mm/sec. The mean volume flow was 3.05xlo-3 cu mm/sec. The mean width of the total peripheral plasma layer was 2. 7 microns and its ratio to mean vessel diameter was 1:11.5. In 50 venules (diameters from 20 to 50 microns), the mean velocity was 1.55 mm/sec. and the mean volume flow rate, 1.76x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 6.7 microns and its ratio to mean vessel diameter was 1:5.3. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.3 and 1:1.7 respectively. A total of 109 mesenteric vessels were studied in frogs anesthetized with urethane. In 55 arterioles (diameters from 15 to 45 microns), the mean velocity was 3.28 mm/sec. The mean volume flow was 2.87x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 3.2 microns and its ratio to mean vessel diameter was 1:9.3. In 54 venules (diameters from 20 to 50 microns), the mean flow rate was 1.61 mm/sec. and the mean volume flow, 1.83x10-3 cu mm/sec. The mean width of the total peripheral plasma layer was 5.6 microns and its ratio to mean vessel diameter was 1:5.9. The ratios of mean arterial to mean venous velocity and volume flow were 1:2.0 and 1:1.5 respectively. In these three series of studies in the frog, no relationship was clearly apparent between velocity and either vessel diameter or width of the peripheral plasma layer in arterioles. Suggestions of a direct relationship between velocity and peripheral plasma layer in veins, however, were evident. In all cases, and in both arterioles and venules, vessel diameter and peripheral plasma layer were clearly and directly related. The effects of flow velocity changes on width of the total peripheral plasma layer in individual vessels were studied in 26 arterioles of the hamster cheek pouch. Blood flow was varied by means of a cuff described by Copley (Biorheology 1: 3, 1962). A direct relationship between velocity and width of the peripheral plasma layer was clearly demonstrated. Thrombus thresholds were determined in mesenteric vessels of the frog. In 103 arterioles (diameters from 20 to 140 microns), the mean strength of stimulus necessary to produce a platelet thrombus was 24.8 volts with an amperage of 0.18 milliamperes. In 100 venules (diameters from 20 to 140 microns), the mean strength of stimulus necessary was 20.7 volts with an amperage of 0.12 milliamperes. Possible relationships of thrombus thresholds to flow velocity in mesenteric arterioles and venules are discussed. / 2031-01-01