The use of diatom assemblages to determine paleotrophic changes in lakes

The purpose of this study was to develop a classifi cation scheme for l ake trophic status based on the relative abundance of l ake sediment diatom trophic indicator species. A total of 600 diatom frustules were counted from the surface sediments of e a ch of 30 lakes selected to repr e seni~ a continuum from u.lt ra-oligotrophic t,o fairly eutrophic but not hype r-' eutrophic conditions. Published autecological information was used to determine the trophic indicator status of each of the s pecies. A quotieht was derived by dividing the s um of all the e utrophic indicator species by the sum of all oligotrophic indicai.-:.or species. Oligo'- mesotrophic. mesotrophic and meso-eutrophic species were added to both the numerator and denomina tor. Five categories of diatom i.nferred trophic status were recognized : ultra-oligotrophic - 0'-0.2:3, oligotrophic::: 0.24-0 . 70, mesotrophic :: 0.'71 -0.99, meso-elxtrophic :: 1. 00-1. '78 and eutrophic:: 1.. 79-2. 43. But only three of these (oligotrophic:: 0-0.69, mesotrophic ::: 0 . 70'-1.69 j and eutrophic:: 1.70-2.50) proved usef ul. The present study of the relationship between diatom inferred trophic status and the literature-derived trophic status of SO lake s (which were purposely chosen to represent a broad spectrum of lakes in Canada) indicated that: 1) Based on diatom species (assemblages ) it is possible to segregate the lakes from which. th",)se diatoms were taken into three basic categories : o ligotrophic, mesotrophic and eutrophic lake types. ~~) It was not possible t,o separate meso-eutrophic and o l igo-mesotrophic lakes f rom mesotrophic l akes as the the degree of overlap betwee n the diat,om species in these lake types was extremely high. 3) Ha d mo r e ul tra-oligotroph,ic lakes been sampled it might have been possible to more a ccurately s eparate them f rom oligotrophic Jakes. 4 ) Had. more humic lakes been sampled in this s tudy I f eel it would have been possible to identify a unique diatom a ssemblage which would h a ve chara cterized t his lake type . Re gression analyses were performed using the aforementioned diatom inferred trophic index as a f unction of 1) log Sec chi transparency (r = - 0.70) 2) total phosphorus (r = 0. 77 ) and 3) chlorophyll-a (r = 0.74). Once e ach of these rel ationships had been established , it was possible to infer paleotrophic (downcore) changes in an oligotrophic lake (Barbara Lake) and in a eutrophic lake (Chemung Lake) . Barbara Llake was dominated by oligotrophic s pecies and remained oligotrophic throughout the 200-·year history r epresented by i t s 32 em long sediment core. Chemung Lake is currently dominated. by eutrophic species but went through a mesotrophic st,age which was associated with a rise in the water level of the lake followi n g dam construction in its watershed in the early 1.900 J ::;. This was followed by its reversion to it,s present day eutrophic stage (dominated by eutrophic species) possibly as a r esult of shallowing process which can be attributed to " silting' up" of the reservoir and the invasion of the l ake by Myriophyllum spjcatum (Eurasian milfoil) i n the 1970's . In addition, nutrient .:r':l.ch run"'offwhich resulted from increased human a.ctivities associated with cottage development along its shores has contribut ed to its eutrophication. There is some evidence that the rat,e o :f its prog ressive eutrophication has declined during the last decade. This was correlated with legislation enacted in the 60's and 70's in Ontario which was aimed at reducing nutrient loading from cottages.

Bioindication of ozone using milkweed plants in Southern Ontario

Ambient (03) ozone concentrations were compared to ozone damage on milkweed plants to determine if there was a correlation. Eight survey sites of at least 100 plants each were located within 5 kilometers of Air Quality Index (AQI) stations in southern Ontario. Sites were visited nine times from June-September (2007) and milkweed leaves from 75 plants were assessed using methods pioneered in the United States. Ambient 0 3 results were calculated into SUM65, seasonal cumulative 0 3, and total 03. The 0 3 exposure indices SUM65 and cumulative 0 3 were tested statistically to determine which index is biologically relevant to milkweed as an 0 3 damage indicator species. The milkweed damage indices were incidence of leaves damaged per plant, incidence of plants damaged per site, and total 0 3• The incidence of plants injured per site was the best damage parameter with an F(1,28)=17.37, p=0.0003 for SUM65 and F(1,28)=7.5, p=O.0106 for cumulative 03 .. Milkweed plants showed quantifiable ozone damage with minimal spatial differences in damage and thus have potential use as a biomonitor species in southern Ontario.

Plant rhizosphere specificity and variability in the insect and plant adhesins, Madl and Mad2, within the genus Metarhizium suggest plant adaptation as an evolutionary force

Metarhizium is a soil-inhabiting fungus currently used as a biological control agent against various insect species, and research efforts are typically focused on its ability to kill insects. In section 1, we tested the hypothesis that species of Metarhizium are not randomly distributed in soils but show plant rhizosphere-specific associations. Results indicated an association of three Metarhizium species (Metarhizium robertsii, M. brunneum and M. guizhouense) with the rhizosphere of certain types of plant species. M. robertsii was the only species that was found associated with grass roots, suggesting a possible exclusion of M. brunneum and M. guizhouense, which was supported by in vitro experiments with grass root exudate. M. guizhouense and M. brunneum only associated with wildflower rhizosphere when co-occurring with M. robertsii. With the exception of these co-occurrences, M. guizhouense was found to associate exclusively with the rhizosphere of tree species, while M. brunneum was found to associate exclusively with the rhizosphere of shrubs and trees. These associations demonstrate that different species of Metarhizium associate with specific plant types. In section 2, we explored the variation in the insect adhesin, Madl, and the plant adhesin, Mad2, in fourteen isolates of Metarhizium representing seven different species. Analysis of the transcriptional elements within the Mad2 promoter region revealed variable STRE, PDS, degenerative TATA box, and TATA box-like regions. Phylogenetic analysis of 5' EF-Ia, which is used for species identification, as well as Madl and Mad2 sequences demonstrated that the Mad2 phylogeny is more congruent with 5' EF-1a than Madl. This suggests Mad2 has diverged among Metarhizium lineages, contributing to clade- and species-specific variation. While other abiotic and biotic factors cannot be excluded in contributing to divergence, it appears that plant associations have been the driving factor causing divergence among Metarhizium species.

FT-IR and MAS NMR analysis of montmorillonite K10 supported MF2 reagents and their activity as catalysis /

ZnF2, CdF2, and CUF2 have been adsorbed onto the surface of montmorillonite K10, and the infrared and 19F, 27 AI, and 29Si MAS NMR spectra of the reagents over a range of loadings have been obtained. CUF2 was observed to attack the Si02 layer and form the complex CuSiF6, Zn F2 tends to attack the aluminium oxide layer, in which Zn isomorphously replaces AI, and forms AIF3 and AIF4 - complexes. All the spectroscopic evidence ruled out the formation of any AI-F and/or Si-F free species as CdF2 is adsorbed on the surface of montmorillonite K10. The reactivity of MF2-K10 reagents towards Friedel-Crafts benzylation of benzene with benzyl chloride varied from one reagent to another. ZnF2-K10 was observed to be the most reactive and CUF2 was the least reactive.

Isolation and characterization of the female sex pheromone of the sugar beet cyst nematode, heterodera schachtii, and an analysis of male precopulatory behaviour /

The sugar beet cyst nematode, Heterodera schachtii, is a major agricultural pest. The disruption of the mating behaviour of this plant parasite in the field may provide a means of biological control, and a subsequent increase in crop yield. The H. schachtii female sex pheromone, which attracts homospecific males, was collected in an aqueous medium and isolated using high performance liquid chromatography. Characterization of the male-attractive material revealed that it was heat stable and water soluble. The aqueous medium conditioned by female H. schachtii was found to be biologically active and stimulated male behaviour in a concentration dependent manner. The activity of the crude pheromone was specific to males of H. schachtii and did not attract second stage juveniles. Results indicated that vanillic acid, a putative nematode pheromone, is not an active component of the H. schachtii sex pheromone. Male H. schachtii exhibited stylet thrusting, a poorly understood behaviour of the male, upon exposure to the female sex pheromone. This behaviour appeared to be associated with mate-finding and was used as a novel indicator of biological activity in bioassays. Serotonin, thought to be involved in the neural control of copulatory behaviour in nematodes, stimulated stylet thrusting. However, the relationship between stylet thrusting induced by the sex pheromone and stylet thrusting induced by serotonin is not clear. Extracellular electrical activity was recorded fi-om the anterior region of H. schachtii males during stylet thrusting, and appeared to be associated with this behaviour. The isolation of the female sex pheromone of H. schachtii may, ultimately, lead to the structural identification and synthesis of the active substance for use in a novel biological control strategy.

Analysis of the feeding behaviour of the mosquito Culex pipiens L. (Diptera: Culicidae) in relation to West Nile virus

The goal ofthis literature review is to inform the reader on several aspects of West Nile Virus (WNV) transmission by its mosquito vector, Culex pipiens and to elucidate how Cx. pipiens and WNV are intertwined. The first few sections of the literature review describe the life cycle and blood feeding behaviours ofmosquitoes so that baseline data ofmosquito biology are established. In addition to explaining how and why a mosquito blood feeds, the section on "Blood Meal Analysis" describes the different methods for determining the vertebrate source of mosquito blood meals and a brief history of these testing methods. Since this thesis looks at the feeding behaviour of Cx. pipiens, it is important to know how to determine what they are feeding upon. Discussion on other mosquito-borne diseases related to WNV gives a broader perspective to the thesis, and examines other diseases that have occurred in Ontario in the past. This is followed by background information on WNV and theories on how this virus came to North America and how it relates to Cx. pipiens. The final sections discuss Cx. pipiens and give background information to how this species of mosquito exists and behaves within North America.

Epidemiology and phylogenetic analysis of West Nile virus in Ontario, Canada, 2002-2003 /

Since the discovery of West Nile (WN) virus in the Western Hemisphere many surveillance programs have been implemented to monitor the epidemiology and genetic variation of WN virus in North America. This project was based on the WN virus Adult Mosquito Identification and Diagnostic Program conducted at Brock University for Ontario, Canada, during the 2002 and 2003 transmission seasons. There are three sections to this thesis. The first section investigated which mosquito species carry WN virus in Ontario, Canada throughout the 2002-2003 transmission seasons. It was found that from the 2002 data, eight mosquito species were detected with WN virus (Aedes vexans, Anopheles punctipennis, Coquilleltidia perlurbans, Culex salinarius, Cx. pipiens, Cx. resluans, Ochlerolalus Irivillalus and Och. Iriserialus) and 7.19% of the total mosquito pools tested were found to be WN virus positive (129 positive poolsll, 793 total pools tested). In 2003, WN virus was detected in only five mosquito species (Ae. vexans, Cx. salinarius, Och. Iriserialus, Cx. pipiens and Cx. resluans) and 1.42% of the total mosquito pools tested were WN virus positive (101 positive poolsl7,1 01 total pools tested). WN virus positive mosquito pools were detected 3-4 weeks earlier in 2002 compared to 2003 data. The second section investigated the actual infection rate (IR) of clearly identified Cx. pipiens and Cx. resluans from the 2002 outbreak. It was found that significantly more ex. resluans were infected with WN virus compared to ex. pipiens. The third section investigated the degree of variability of the WN virus genome. A 879 nucleotide section of the WN virus genome was amplified from 21 American Crows and 20 adult female mosquitoes from Ontario, Canada, and compared to the homologous region of the original New York 1999 Chilean Flamingo sequence (NY99FL). Seventy-two nucleotides from Ontario WN virus sequences showed variability compared to NY99FL with 10 synapotypic changes. Phylogenetic analysis revealed a close relationship between Ontario and US WN virus sequences.

A dimensional analysis of the benzylic hydroxylase active site in mortierella isabellina

The spatial limits of the active site in the benzylic hydroxylase enzyme of the fungus Mortierella isabellina were investigated. Several molecular probes were used in incubation experiments to determine the acceptability of each compound by this enzyme. The yields of benzylic alcohols provided information on the acceptability of the particular compound into the active site, and the enantiomeric excess values provided information on the "fit" of acceptable substrates. Measurements of the molecular models were made using Cambridge Scientific Computing Inc. CSC Chem 3D Plus modeling program. i The dimensional limits of the aromatic binding pocket of the benzylic hydroxylase were tested using suitably substituted ethyl benzenes. Both the depth (para substituted substrates) and width (ortho and meta substituted substrates) of this region were investigated, with results demonstrating absolute spatial limits in both directions in the plane of the aromatic ring of 7.3 Angstroms for the depth and 7.1 Angstroms for the width. A minimum requirement for the height of this region has also been established at 6.2 Angstroms. The region containing the active oxygen species was also investigated, using a series of alkylphenylmethanes and fused ring systems in indan, 1,2,3,4-tetrahydronaphthalene and benzocycloheptene substrates. A maximum distance of 6.9 Angstroms (including the 1.5 Angstroms from the phenyl substituent to the active center of the heme prosthetic group of the enzyme) has been established extending directly in ii front of the aromatic binding pocket. The other dimensions in this region of the benzylic hydroxylase active site will require further investigation to establish maximum allowable values. An explanation of the stereochemical distributions in the obtained products has also been put forth that correlates well with the experimental observations.

Environmental assessment and biomonitoring of the Twelve Mile Creek watershed, Niagara Peninsula, Ontario

In light of the heavy reliance of the people of the Niagara Peninsula on the T\\'elve Mile Creek (TMC) watershed for recreational activities and for municipal and industrial uses ( e.g., drinking water, shipping and discharge of effluents), it was deemed prudent to assess the envirol1tnental health of the system by analysing the sediments total and exchangeable metal, and TPH contents. The MOEE has set guidelines with limits for the protection and management of aquatic sediments, and the sediments from the headwaters of the TMC have total metal and TPH (subset of O&G) contents well below the lower provincial limits. Areas of environmental concern where total metal contents in sediments, either individually or collectively, exceed the guideline, are the south side of Lake Gibson, the Old WeIland Canal, a segment of TMC just south of the QEW and Martindale Pond. The total metal content of sediments does not in all instances identify areas of biological concern. Instead, it has been found that the exchangeable metal fraction of sediments is a better indicator of metal availability and thus potential accumulation in organisms. In some instances, the exchangeable metal fraction agrees with the total metal fraction defining areas of environmental concern, but it does vary from site to site reflecting the natural variability of the ambient environment. Overall, the exchangeable metal fraction of sediments appears to be a better indicator of anthropogenic pollution and ecosystem impact. A histochemical study of Anodon.ta sp., Elliptio sp. and zebra mussels (Dreissena polyn'101pha) was done in conjunction with passive biomonitoring of zebra and quagga mussels (Dreissena bugensis) from the Twelve Mile Creek watershed and Lake 51. Clair (Jeanette's Creek, Chatham, Ontario). The highest concentrations of divalent metals such as Cu, Ni, Cd, and Zn, and trivalent Al appear to accumulate in gill and kidney tissues. Metal contents of organ tissues in Anodonta sp. vary with size class. Organ metal content varies among size classes, thus requiring consideration of size in biomonitoring studies. Shucked zebra and quagga mussel tissues, exhibited similar size class to Al content trends. In addition they reflected the Al content trends of top (approximately 10 cm) most sediments in the Twelve Mile Creek watershed. Quagga mussels appear to have higher Al concentrations than zebra mussels, thus suggesting that quagga mussels may be better passive biomonitors of AI. Cd content in zebra mussel tissues, seemed to increase with size class trends. This was not demonstrated in the quagga mussel tissues. This suggests that Cd may be regulated by quagga mussels and not by zebra mussels, and that zebra mussels may be better passivebiomonitors of Cd than are quagga mussels. Zebra mussel, quagga mussel, Anodonta sp., and Elliptio sp. were used in a two part, active (translocated) biomonitoring study of the Twelve Mile Creek watershed. There was no statistical difference in death rates between zebra and quagga mussels after 65 days of biomonitoring. However there does appear to be a difference of death rates between sites. Unfortunately the data base did not permit us to differentiate between sites. Relative to Port Colborne Harbour (Port Colborne, Ontario), the Twelve Mile Creek watershed appears to be elevated in bioavailable AI. An area near the terminus of the Twelve Mile Creek appears to be an area of environmental concern since mussels seemed to have accumulated relatively large concentrations of Cd, Zn, and Pb. In addition to possible metal loading from a nearby outfalls, or possible upstream outfalls, road salt runoff from storm sewers may have contributed to metal accumulation through cation exchanges processes. Similar trends in cumulative quagga mussel metal concentrations during the two time periods (65 and 159 days), suggest that quagga mussels may reach equilibrium within 65 days of translocation. Differences in bioaccumulated metal concentrations of the two dreissenid species demonstrate that active biomonitoring studies must use a variety of organisms to adequately assess the environmental situation of specific waterways and/or bodies.

Microstructural and geochemical investigation of North American lower-middle Paleozoic trilobites and recent arthropods

Trilobites ¥tere collected from Ordovician and Devonian formations of Ontario} New York} Ohio} Oklahoma} and Indiana. Diversity was generally low} but 19..?telllS and Ph..~tY>ps ¥tere the most abundant species from the Ordovician and Devonian} respectively. Recent marine arthropods ¥tere collected from the Atlantic shore of the middle Florida Keys} and from the Pacific and lagoonal waters at Cape Beale} B. C. Fresh-water arthropods were collected along the shore of the Severn River in northcentral Ontario. Cuticles ¥tere analyzed for major} minor and trace elements, 180 and 13C isotopes, as ¥tell as examined by scanning electron micr?scope to identify original and diagenetic fabrics. Examination of trilobite cuticles by scanning electron microscope revealed several microstructures consistent with those observed in Recent arthropods. Microstructures} such as setae and tegumental gland duct openings} in like sized Lim/IllS and Isoteline trilobites may indicate common ancestral origins for these organisms, or simply parallel cuticle evolutions. The dendritic microstructure, originally' thought to be a diagenetic indicator, was found in Recent specimens and therefore its presence in trilobites may be suggestive of the delicate nature of diagenesis in trilobites. The absence of other primary microstructures in trilobites may indicate alteration, taxonomic control} or that there is some inherent feature of S EM examination which may' not allow detection of some features} while others are apparently visit·le onl~1 under SH.·1. The region of the cuticle sampled for examination is also a major influence in detecting pristine microstructures, as not all areas of trilobite and Recent arthropod cuticles will have microstructures identifiable in a SEM study. Subtleties in the process of alteration, however} ma~·· leave pristine microstructures in cuticles that are partial~/ silicified or do 10m itized, and degree and type of alteration may vary stratigraphically and longitudinally within a unit. The presence of fused matrices, angular calcite rhombs, and pyrite in the cuticle are thought to be indicative of altered cuticles, although pyritization may not affect the entire cuticle. t-~atural processes in Recent arthropods, such as molting, lead to variations in cuticle chemistries, and are thought to reflect the area of concentration of the elements during calcification. The level of sodium in Recent arthropods was found to be higher than that in trilobites, but highly mobile when sUbjected to the actions of VY'€'athering. Less saline water produced lovy'€'r magnesium and higher calcium values in Recent specimens .. and metal variations in pristine Ordovician trilobite cuticle appears to follow the constraints outlined for Recent arthropods, of regulation due to the chemislry of the surrounding medium. In diagenetic analysis, sodium, strontium and magnesium proved most beneficial in separating altered from least altered trilobites. Using this criterion, specimens from shale show the least amount of geochemical alteration, and have an original mineralogy of 1.7 - 2.4 mole % MgC03 (8000 t(> 9500 ppm magnesium) for both /s>..?/e/11S lJA'i.riff!11S and PseIAit'11J17ites I..itmirpin..itl/~ and 2.8 - 3.3 mole % MgC03 (5000 to 7000 ppm magnesium) for Ph.i{).?PS This is Slightly lower than the mineralogy of Recent marine arthropods (4.43 - 12.1 mole % MgC03), and slightly higher than that of fresh-water crayfish (0.96 - 1.82 mole % MgC03). Geochemically pristine trilobites were also found to possess primary microstructures. Stable isotope values and trends support the assertion that marine-meteoriclburial fluids were responsible for the alteration observed in a number of the trilobite specimens. The results of this stUdy suggest that fossil material has to be evaluated separately along taxonomic and lithological lines to arrive at sensible diagenetic and e nvironmenta I interpretations.

The utility of sedimentary diatoms as a measure of historical lake ph

As a result of increased acid precipitation, the pH of a large number of Canadian Shield lakes has been falling. Prior to this study there was no documentation available to explain the history of lake acidification for the Algoma area lakes. In order to obtain this information the diatom inferred pH technique was developed in this study. During two field seasons, July 1981 and July 1982, short sediment cores (circa 25-30 cm) were collected from 28 study lakes located north of Lake Superior, District Algoma, Ontario. The surface sediment diatoms (0-1 cm) from each of these lakes were carefully identified, enumerated, and classified in terms of their pH indicator status. The surface sediment diatom analysis indicated that lake pH is one of the most important factors affecting the species composition and relative abundance of diatom populations. Thus diatom assemblages can be sensitive indicators of lake acidification. When Nygaard's index alpha was plotted against observed lake pH, a statistically significant relationship resulted (r=-0.89; p=indicators of lake pH. Thus diatom inferred pH technique has great potential in explaining the rate of lake acidification.

"I might be a duck, but I'm human" : an analysis of clothing in Disney cartoons

Mickey Mouse, one of the world's most recognizable cartoon characters, did not wear a shirt in his earliest incarnation in theatrical shorts and, for many years, Donald Duck did not wear pants and still rarely does so. Especially when one considers the era in which these figures were first created by the Walt Disney Studio, in the 1920s and 1930s, why are they portrayed without full clothing? The obvious answer, of course, is that they are animals, and animals do not wear clothes. But these are no ordinary animals: in most cases, they do wear clothing - some clothing, at least - and they walk on two legs, talk in a more or less intelligible fashion, and display a number of other anthropomorphic traits. If they are essentially animals, why do they wear clothing at all? On the other hand, if these characters are more human than animal, as suggested by other behavioral traits - they walk, talk, work, read, and so on - why are they not more often fully clothed? To answer these questions I undertook three major research strategies used to gather evidence: interpretive textual analysis of 321 cartoons; secondary analysis of interviews conducted with the animators who created the Disney characters; and historical and archival research on the Disney Company and on the times and context in which it functioned. I was able to identify five themes that played a large part in what kind of clothing a character wore; first, the character's gender and/or sexuality; second, what species or "race" the character was; third, the character's socio-economic status; fourth, the degree to which the character was anthropomorphized; and, fifth, the context in which the character and its clothing appeared in a particular scene or narrative. I concluded that all of these factors played a part in determining, to some extent, the clothing worn by particular characters at particular times. However, certain patterns emerged from the analysis that could not be explained by these factors alone or in combination. Therefore, my analysis also investigates the individual and collective attitudes and desires of the men in the Disney studio who were responsible for creating these characters and the cultural conditions under which they were created. Drawing on literature from the psychoanalytic approach to film studies, I argue that the clothing choices spoke to an idealized fantasy world to which the animators (most importantly, Walt Disney himself), and possibly wider society, wanted to return.

Spectral Reflectance Characteristics of Kimberlite Indicator Minerals and Other Selected Mineral Grains

High chromium content in kimberlite indicator minerals such as pyrope garnet and diopside is often correlated with the presence of diamonds. In this study, kimberlite indicator minerals were examined using visible light reflectance spectroscopy to determine if chromium content can be correlated with spectral absorption features. The depth of absorption features in the visible spectral region were correlated with the molecular percentage of chromium and other first series transition metal elements obtained by electron microprobe data. In the visible part of the spectrum, chromium is evident by 3 absorption features in the pyrope reflectance spectrum; one isolated and narrow feature at the wavelength 689 nm was used to correlate with the chromium mol %. The isolation of this feature in the pyrope spectra is advantageous since it is not directly affected by other proximal absorption bands that could be caused by other transition metals. Analysis of the feature indicates that as grain volume increases the depth of the absorption feature will also increase. Clustering grain volumes into fractions yields better correlation between absorption depth and mol % chromium. Other types of garnet (almandine, grossular, spessartine) and kimberlite indicator minerals (olivine, diopside, chromite, ilmenite) were analyzed to determine if other absorption features could be used to predict the proportion of specific transition metal elements. Diopside in particular illustrates the same isolated chromium absorption feature as pyrope and may indicate mol percent but needs further study with larger sample sets.