Invasive colonial tunicates pose substantial economic threat to the shellfish aquaculture industry, but their population dynamics and ecological impacts are highly variable and region-specific. This thesis contributes to our regional understanding of two such tunicates in Oregon. The first chapter explores the population dynamics of Didemnum vexillum, one of Oregon’s top 100 most dangerous invasive species, at an oyster farm. From May 2011 to 2016 the population fluctuated extensively, though did not exhibit any net growth over the study period. In the second chapter, I demonstrate that Botrylloides violaceus had no impact on the growth, condition, or organic composition of oysters and mussels grown in a simulation of longline aquaculture. Together, these studies paint a cautiously positive outlook for the shellfish aquaculture industry in Oregon. This thesis includes previously unpublished co-authored material.

No description available.

Raman spectroscopy has been used to monitor the deformation of natural cellulose whisker polymer nanocomposites. Cotton and tunicate whiskers were used as reinforcements in polymer matrices. Raman spectra from the nanocomposites highlight an intense band located at the 1095 cm-1 position. This band is reported to shift towards a lower wavenumber under the application of tensile deformation. On the other hand, the compressive deformation of the composite gives rise to an increase in the position of this Raman band. The shifts correspond to the direct deformation of the molecular backbone of cellulose, which is dominated by a C-O stretching mode. The Raman band located at 1095 cm-1 is shown to shift non-linearly before it reaches a plateau due to the breakdown of the whisker-matrix interface. The initial shift rate is associated with the stiffness of the cellulose whiskers. The stiffnesses of single whiskers of cotton and tunicate are found to be 58 and 155 GPa respectively, assuming two dimensional (2D) in-plane distribution of whiskers. Cyclic deformation tests of the composites provide an insight into understanding the behaviour of the whisker-polymer matrix interface under tension and compression. It is found that residual compressive stress occurs during each cycle of the deformation. The level of disruption at the whisker-matrix interface is determined by estimating the energy dissipation, which is proportional to the hysteresis area. Local orientation is also observed in the nanocomposites produced by solution casting and subsequent melt pressing. Dark regions of the composites viewed under a polarised optical microscopy are found to represent areas in which the cellulose whiskers form a randomly oriented whisker network. A shift rate for the Raman band initially located at 1095 cm-1 obtained in the dark regions of 12.2 vol% tunicate whisker poly(vinyl acetate) nanocomposites is found to be -0.5±0.07 cm-1%-1, which is lower than -1.2±0.04 cm-1%-1 from the bright regions. Exposure to water and temperature during the deformation of the nanocomposites results in significant changes in stress transfer between the whiskers and the matrix. It is shown that the interface can be 'switched-off' for the poly(vinyl acetate)/whisker system in the presence of water and also at temperature above the glass transition.

Of the small percentage of organisms chemically investigated over the years as potential sources of natural products, much less is known about those from the marine realm. Despite the lack of attention they have received in comparison to terrestrial organisms, marine life have recently been found to represent a valuable source for novel bioactive compounds. Cold water marine habitats are home to a plethora of organisms that have the ability to produce secondary metabolites that exhibit a great deal of diversity in both their chemical structures and biological activities. The chemical investigation of these unique and relatively unstudied ecosystems is necessary to gain insight into the dynamics between predators and prey, while also making a significant impact in the field of drug discovery. Our laboratory has focused on the chemical investigation of invertebrates from the waters of Antarctica in search of bioactive secondary metabolites that can be used for the treatment of human pathogens. This dissertation reports a small portion of the progress made in our laboratory towards the exploration of Antarctic marine invertebrates. The chemical investigation of the circumpolar colonial tunicate Synoicum adareanum and the orange, encrusting sponge Artemisina plumosa will be discussed in detail in the following chapters.

Synoicum adareanum is a colonial tunicate commonly found on the benthos around Palmer Station on Anvers Island, Antarctica. A comprehensive chemical investigation of the lipophilic extract of the frozen tunicates gave a new series of polyketide macrolides, palmerolides A-E and H. The structure elucidation of these compounds was accomplished by extensive NMR and mass spectral studies.The palmerolides are unusual 20-membered macrolides displaying functionality more commonly found in sponges or cyanobacteria. Palmerolide A, the major member of the group, shows significant and selective in vitro cytotoxicity against melanoma with three orders of greater sensitivity relative to other cell lines tested, in the National Cancer Institute (NCI) 60 human cancer cell-line panel. In addition, it displays potent cytostatic activity against several other cancer cell lines. Based on NCI's COMPARE analysis, palmerolide A was investigated as a V-ATPase inhibitor and shown to bind the V0 subunit with 2 nM inhibition.Austrodoris kerguelenensis is a common Antarctic nudibranch widely distributed in the High Antarctic and Sub Antarctic Zone. It is characterized by the presence of terpenoid glyceryl esters which are supposed to be involved in defense. Chemical investigations of several specimens of A. kerguelenensis collected near Palmer station Antarctica afforded hitherto undescribed series of clerodane diterpenoid glycerides. The structure elucidation of three major compounds of this series, palmadorin A, B and C was accomplished.

Marine ecosystems can be a promising reservoir of various kinds of chemical components, applicable as pharmaceutical materials, food, cosmetics, nutraceuticals, and others for different industry. As an example, Tunicates, a group of marine animals, have been attracted a lot of attention in medical application, food market, water pollution issues, and Cellulose nanomaterial production due to their consisting of chemical compounds such as cellulose, amino-sugars, and proteins or protein-polysaccharide complexes e.g. collagen, glycosaminoglycan, chitin, scleroprotein, iodine-binding proteins, and elastin. In this project,  two dominant species of Scandinavian Tunicates, i.e. Ciona intestinalis and Clavelina lepadiformis, harvested from Norwegian ocean have been classified according to body sizes, depths from the ocean surface, ages and species, and separated physically into outer layer and internal organs, followed by measurements of sugar composition, oil content, and  protein content. Application potentials have been investigated by trials for production of pure crystalline cellulose, bioethanol, and biodiesel, and by analysis of amino acid composition of the samples. The cellulose percentage and cellulose yield for the chemically pure cellulose obtained, is around 96% and 54% respectively, and the protein content is decreased step by step by the acid, alkali, and bleaching process applied. Bioethanol can be obtained by fermentation of tunicate hydrolysate with strains A and C which are derived from Saccharomyces cerevisiae. The biodiesel yield of tunicate samples is around 4-6% as an average. The amino acid compositions in our tunicate samples are similar to egg albumin, implying tunicate being an alternative material for animal feed production. Several processing treatments have been conducted with the aims to fractionate tunicate biomass components or enhance the cellulose accessibility and reactivity. After a single processing step, Ba(OH)2 treated samples seemed to be the best in terms of both cellulose preservation (66.5% cellulose) and protein removal (6% protein in the treated residue). Results from the physical separation plus washing reveal that the highest amount of cellulose and protein presents is found in the outer (Tunic) part and internal organs of Tunicate samples respectively. Data obtained from FTIR(Fourier Transform Infrared Spectroscopy) and SEM(Scanning Electron Microscope) indicate that among all processing trials, H3PO4 is the most effective in decreasing the cellulose crystallinity, which renders a higher accessibility for acidic or enzymatic reaction during bioethanol production due to a higher amount of amorphous structure of cellulose. From the analysis results of component contents and structures, it could be concluded that increase of deepness results in a decrease of sugar content of the Tunicate samples while there are no differences in protein and carbohydrate content in different tunicate species. The body size has a positive influence on the protein content and the sample age alters the contents of both sugar and protein. In addition, Tunicate oil has high phospholipid content instead of glycerol ester, the latter being the common oil from vegetable origins. Moreover, lots of free fatty acid is present, and the composition profile of Tunicate fatty acids seems to be similar to fish oil, as revealed by NMR (Nuclear Magnetic Resonance Spectroscopy), FTIR, and GC-MS (Gas Chromatography-Mass Spectrometry).

Marine ecosystems can be a promising reservoir of various kinds of chemical components, applicable as pharmaceutical materials, food, cosmetics, nutraceuticals, and others for different industry. As an example, Tunicates, a group of marine animals, have been attracted a lot of attention in medical application, food market, water pollution issues, and Cellulose nanomaterial production due to their consisting of chemical compounds such as cellulose, amino-sugars, and proteins or protein-polysaccharide complexes e.g. collagen, glycosaminoglycan, chitin, scleroprotein, iodine-binding proteins, and elastin. In this project,  two dominant species of Scandinavian Tunicates, i.e. Ciona intestinalis and Clavelina lepadiformis, harvested from Norwegian ocean have been classified according to body sizes, depths from the ocean surface, ages and species, and separated physically into outer layer and internal organs, followed by measurements of sugar composition, oil content, and  protein content. Application potentials have been investigated by trials for production of pure crystalline cellulose, bioethanol, and biodiesel, and by analysis of amino acid composition of the samples. The cellulose percentage and cellulose yield for the chemically pure cellulose obtained, is around 96% and 54% respectively, and the protein content is decreased step by step by the acid, alkali, and bleaching process applied. Bioethanol can be obtained by fermentation of tunicate hydrolysate with strains A and C which are derived from Saccharomyces cerevisiae. The biodiesel yield of tunicate samples is around 4-6% as an average. The amino acid compositions in our tunicate samples are similar to egg albumin, implying tunicate being an alternative material for animal feed production. Several processing treatments have been conducted with the aims to fractionate tunicate biomass components or enhance the cellulose accessibility and reactivity. After a single processing step, Ba(OH)2 treated samples seemed to be the best in terms of both cellulose preservation (66.5% cellulose) and protein removal (6% protein in the treated residue). Results from the physical separation plus washing reveal that the highest amount of cellulose and protein presents is found in the outer (Tunic) part and internal organs of Tunicate samples respectively. Data obtained from FTIR(Fourier Transform Infrared Spectroscopy) and SEM(Scanning Electron Microscope) indicate that among all processing trials, H3PO4 is the most effective in decreasing the cellulose crystallinity, which renders a higher accessibility for acidic or enzymatic reaction during bioethanol production due to a higher amount of amorphous structure of cellulose. From the analysis results of component contents and structures, it could be concluded that increase of deepness results in a decrease of sugar content of the Tunicate samples while there are no differences in protein and carbohydrate content in different tunicate species. The body size has a positive influence on the protein content and the sample age alters the contents of both sugar and protein. In addition, Tunicate oil has high phospholipid content instead of glycerol ester, the latter being the common oil from vegetable origins. Moreover, lots of free fatty acid is present, and the composition profile of Tunicate fatty acids seems to be similar to fish oil, as revealed by NMR (Nuclear Magnetic Resonance Spectroscopy), FTIR, and GC-MS (Gas Chromatography-Mass Spectrometry).

This thesis is about the demography of the tunicate Styela clava, a species of some notoriety because of its invasiveness and impacts in many parts of the world. Species assemblages have continuously changed throughout evolutionary history, but the rate of today’s anthropogenically facilitated dispersal is unparalleled in history. Non-indigenous species (NIS) are now considered one of the most important risks to native biodiversity. NIS become invasive by becoming both widespread and locally dominant. This requires that a species becomes established, spreads locally, and increases in abundance. In the early stages of invasion, its demography and life history characteristics are of crucial importance. In New Zealand, Styela has established populations in several places, but none of these populations has yet reached the high densities found in other countries. In Lyttelton Port, where this study was located, Styela was first noticed in 2005. It therefore presented an ideal situation to study an invasive species in its early stages of establishment and provided a potentially good model for understanding how invasive species get local traction and spread from initial infestation points. Therefore, I set out to determine demographic features of Styela to understand the numbers game of population dynamics. This study used empirical data on growth rates, size-frequencies through time, and size and age to maturity to test several models, including von Bertalanffy, Logistic dose-response, Ricker and power models of individual growth. The most useful proved to be the von Bertalanffy model. Styela individuals shrink frequently, so average growth rates were often quite low, even though some individuals reached 160 mm or more in total length. Mortality was greatest in summer, presumably after reproduction, and lowest in winter. Fewer than 5% of individuals survived 12 months, and most or all of these died soon afterwards. Populations were, therefore, essentially annual. Recruitment was difficult to determine because of the cryptic nature of small juveniles. However, size-frequency, abundance and mortality data indicated that recruitment most likely occurred in early spring (late-October), and then again in late summer, with growth to maturity (at c. 50 mm total length) within < 5 months. Several manipulative experiments showed that Styela did not readily capitalise on provision of free space but the other non-native ascidian, Ciona intestinalis, rapidly recruited. Transplants of Styela were greatly affected by C. intestinalis, which overgrew them, similar to a localised replacement of Styela by Ciona seen overseas. Lefkovitch modelling was used to test whether Styela had an “Achilles heel” in its life history, whereby managed removal could impact future populations. This showed that under several scenarios intervention would most likely be ineffectual. Overall, this study showed that the original populations in Lyttelton Port are either static or in decline, somewhat contrary to original expectations. Nevertheless, it appears that these small populations may be acting as stepping stones for spread of this species outside of the port.

Tunicata umfasst 3000 marine Arten, mit sehr unterschiedlichen Lebensstrategien. Als eines der drei großen Taxa innerhalb der Chordata, stellt die Evolution der Tunikaten eine Schlüsselkomponente bei der Aufklärung der Evolution der Chordaten und Cranioten dar. Dafür ist ein Verständnis der Merkmalstransformationen innerhalb der Tunikaten notwendig. Leider sind die internen Verwandtschaftsverhältnisse der fünf großen Tunikatentaxa in verschiedenen molekularphylogenetischen Studien widersprüchlich. Bisher gibt es nur wenige morphologische phylogenetische Analysen. Ein Schwerpunkt dieser Arbeit liegt auf der Untersuchung neuroanatomischer Merkmale, da das Nervensystem wahrscheinlich phylogenetische Informationen enthält. Durch das Anwenden moderner morphologischer Methoden, wie hochauflösende konfokale Laserscan- und Elektronenmikroskopie (REM und TEM), und 3d Rekonstruktionen basierend auf lichtmikroskopischen Schnitten, wurde die Verfügbarkeit neuroanatomischer Daten wesentlich verbessert. Die Ergebnisse zeigen, dass die Variation neuroanatomischer Merkmale größer ist als bisher angenommen und dass sich die Gehirnanatomie und die Verteilung von Neurotransmittern in den zwei Stadien der Thaliaceen unterscheidet. Neue unabhängige Merkmale des Nervensystems wurden in einer Matrix kodiert. Ergänzt mit traditionellen in der Tunikatentaxonomie verwendeten Merkmalen, entstand die bisher umfangreichste morphologische Datenmatrix, die 116 Merkmale für insgesamt 54 Arten umfasst. Die kladistische Analyse ergab monophyletische Tunicata, in denen die Appendicularia die Schwestergruppe der übrigen Tunikaten bildet. Ascidiacea ist monophyletisch, während „Thaliacea“ paraphyletisch ist. Zusätzlich wurde eine kombinierte phylogenetische Analyse basierend auf den morphologischen Daten und 18S rDNA-Sequenzen durchgeführt. Eine stufenweise stärkere Gewichtung phänotypischer Merkmale zeigt, dass die morphologischen Daten das Ergebnis der kladistischen Analyse stark beeinflussen. / Tunicata comprises 3000 marine species with diverse life-history strategies. As one of the three major chordate taxa, the evolution of tunicates plays a key role to elucidate chordate and craniate evolution. Therefore, a broader understanding of character transformations within tunicates is essential, but the interrelationships of the five main tunicate subtaxa in previous molecular phylogenetic analyses were contradictory. Morphological phylogenetic analyses are rare. In this comparative study emphasis was given to neuroanatomical characters, as the nervous system probably contains phylogenetic information. Applying modern morphological techniques like high-resolution confocal laser scanning microscopy and electron microscopy (SEM and TEM), serial sectioning for light microscopy, and digital 3d reconstruction, the number of available tunicate neuroanatomical data was considerably increased. It was revealed that the variation of neuroanatomical characters is higher than previously assumed, a specific pattern of serotonin-like immunoreactive cells in ascidians is present, and that brain anatomy and distribution of neurotransmitters in the two thaliacean life-cycle stages differs. Novel independent characters of the central nervous system were coded in a matrix for a cladistic analysis. Including traditional morphological from tunicate literature this effort resulted in the largest morphological data matrix to date, containing 116 phenotypic characters and 54 species. The cladistic analysis resulted in monophyletic Tunicata, with Appendicularia the sister taxon to the remaining tunicates. Furthermore, the monophyly of Ascidiacea is supported, whereas “Thaliacea” are paraphyletic. An additional phylogenetic analysis combining morphological and 18S rDNA-sequence data was performed. A reevaluation of this dataset with a successively increased weighting of the phenotypic data showed that morphological data strongly influence the outcome of the cladistic analysis.

There is growing demand for a more efficient use of polymers that originate from renewable feedstocks due to the depleting supply of fossil fuels, based on economic and environmental reasons. As a result, lignin has attracted renewed interest as a resource for various bioproducts. Lignin is a natural biopolymer with a high carbon content and is composed of aromatic moieties, with a high level of polar functionalities. This makes it a unique precursor for certain high-value applications, such as in biofuels, bioplastics, composite materials, carbon fibers and activated carbons and as a source of phenolic monomers and fine chemicals. Industrial lignins are formed as byproducts of pulping processes (such as kraft, sulfite or alkaline pulping) or result from the biorefining process, where carbohydrates are used for sugar production. Lignin’s intrinsic structure is significantly modified during the processing of lignocellulose, resulting in the formation of more diverse, condensed and less reactive raw materials. Since molecular mass and polydispersity are the most important parameters affecting the chemical reactivity and thermal properties of lignin, additional process steps to improve the quality of crude technical lignins, including kraft lignin, are needed. Solvent extraction is a potentially useful technique for further improving the polydispersity of technical lignins. This work summarizes the impact of solvent fractionation on the chemical structure, antioxidant activity, heating value, and thermal and sorption properties of industrial hardwood and softwood kraft lignins. The purpose was to understand the correlation between certain structural features in the lignin fractions and their properties to select the appropriate solvent combinations for specific applications of lignin raw materials. Four common industrial solvents, namely, ethyl acetate, ethanol, methanol and acetone, in various combinations were used to separate both spruce and eucalyptus kraft lignins into fractions with lower polydispersities. Gel-permeation chromatography analysis was used to evaluate the efficiency of the chosen solvent combination for lignin fractionation. The composition and structure of the lignin material were characterized by elemental analysis, analytical pyrolysis (Py-GC/MS/FID) and 31P NMR spectroscopy. The thermal properties of the lignin samples were studied by thermogravimetric analysis. Proximate analysis data (ash, volatile components, organic matter and fixed carbon) were obtained through the direct measurement of weight changes during the analysis, while the high heating values (in MJ/kg) were calculated according to equations suggested in the literature. The sorption properties of fractionated kraft lignins were studied with respect to methylene blue dye. Additionally, lignin fractions with different molecular weights (and therefore various chemical structures) that were isolated from both softwood and hardwood kraft lignins were incorporated into a tunicate cellulose nanofiber (CNF)-starch mixture to prepare 100% bio-based composite films. The aim was to investigate the correlation between lignin diversity and film performance. The transmittance, density and thermal properties of the films were investigated, as were their mechanical properties, including the tensile stress and Young’s modulus. This part of the study addressed the importance of lignin diversity on composite film performance, which could be helpful for tailoring lignin applications in bio-based composite materials based on the material’s specific requirements.