Restos de moluscos acumulados na superfície do sedimento marinho (i.e., assembleias mortas, retrabalhadas ou não) podem ser os principais testemunhos do efeito de fatores ambientais (oceanográficos, sedimentares, climáticos) sobre o modo como os moluscos fósseis são preservados. Ou seja, o quanto o meio ambiente é responsável pela fossilização de restos esqueléticos só pode ser medido, com máxima fidelidade, através do estudo de assembleias modernas que possuam restos refratários (e.g., conchas). Fatores ambientais atuais podem influenciar o dano tafonômico em moluscos de depósitos retrabalhados. Embora muitos estudos atualísticos tenham estabelecido relações entre determinados fatores ambientais e danos causados às conchas, poucos i) enfocam a fração de microclastos (< 4 mm), são comuns em amostras de subsuperfície, ou ii) analisam essas relações utilizando escalas espaciais maiores que local (e.g., regional, diferentes províncias marinhas), especialmente com o propósito de testar certos fatores ambientais em escala local e ampla (e.g., temperatura local vs. faixas de temperatura). Os raros estudos existentes tratam-se meta análises. No registro fóssil, muitas vezes, apenas gradientes ambientais amplos são recuperados. Por isso é importante avaliar a relação destes fatores (modernos) com as assinaturas tafonômicas em moluscos. O objetivo desta tese foi determinar até que ponto variações em fatores ambientais (salinidade, temperatura, velocidade de corrente, granulometria e composição do sedimento) geralmente observados em escala local, influenciam o tipo de dano tafonômico em micro e macroclastos de moluscos marinhos, se analisados em escala mais ampla. Grande parte do material analisado esteve representado por clastos de remaniés existentes na Plataforma Sul Brasileira (PSB). Desta forma, o presente estudo também investigou se fatores ambientais atuais podem ser deduzidos a partir de perfis tafonômicos de assembleias mortas retrabalhadas. Os resultados obtidos mostraram que, na PSB, i) apesar de diferentes em termos absolutos, o perfil de dano de cada amostra analisada não varia entre macro e micromoluscos, e que ii) quando o estudo é conduzido em escala espacial mais ampla, até 60% da variação no perfil tafonômico pode ser explicada pela variação ambiental. Ainda nesta região, os principais fatores ambientais que afetaram o dano em microclastos de bivalves são: i) salinidade, que reflete o aporte de água doce (e da circulação oceânica como um todo), e ii) proporção de carbonato no sedimento, que reflete, primariamente, a existência de depósitos bioclásticos (constituídos de conchas de moluscos retrabalhadas, ou remaniés biológicos). Os depósitos bioclásticos mais proximais (ao sul da desembocadura da Lagoa dos Patos) apresentaram maior evidência de dissolução e retrabalhamento físico, enquanto que os bioclastos do depósito mais distal (entre as profundidades de ~ 100 e 200 m) apresentaram melhor preservação tafonômica. Além disso, também pôde ser determinado que a alteração de cor dos bioclastos na PSB (bastante ubíquo em amostras concentradas em cascalho e carbonato) está vinculada, significativamente, a estados de oxidação/redução do sedimento. Estes, por sua vez, estão relacionados à presença de elementos metálicos (como ferro e manganês), importantes na mineralização do carbono de origem biológica, e na geoquímica da zona tafonomicamente ativa, em ambiente marinho. Na análise conduzida em escala espacial geográfica, incluindo material do Plataforma do Nordeste do Brasil, pode ser determinado que profundidade, um fator ambiental com influência significativa sobre o perfil de dano na PSB, perde importância. Em diferentes províncias geográficas, os principais fatores que influenciaram as assinaturas tafonômicas estão vinculados à velocidade da corrente, salinidade e proporção de carbonato no sedimento. Estes refletiram a influência de diferentes massas d‟água (Corrente Norte do Brasil e Corrente das Malvinas) e diferentes regimes de sedimentação (siliciclástica relíquia e carbonática atual). / Dead molluscan remains (i.e., death assemblages, reworked or not), accumulated on the marine sedimentary surface, can preserve most of the environmental factors (oceanographic, sedimentary, climatic) which control fossil mollusk preservation. That is, how much the environment is responsible on skeletal remain fossilization can only be measured, with the highest fidelity, by studying modern assemblages possessing refractory parts (such as shells). Present-day environmental factors can influence the taphonomic damage on reworked mollusk remains. Although atualistic studies had constrained some environmental factors and shell damage, a few i) focuses mesh sizes lower than 4 mm (microclasts), which are common in subsurface samples, or ii) analyze these relationships using spatial scales larger than local (e.g., regional, different marine provinces), especially with the explicit purpose of to test some environmental factors on local and large scale (e.g., local temperature vs. temperature ranges). The rare studies available are meta-analysis. In the fossil record, sometimes only large environmental gradients can be recovered. That is why it is important to evaluate the relationship between modern environmental factors and taphonomic signatures in mollusks. The subject of this thesis is to determine to which degree the variation in environmental factors (salinity, temperature, current velocity, sediment granulometry and composition), usually observed in local scale, do influence the kind of taphonomic alteration in micro and macroclasts of marine mollusks, when analyzed in large scale. Most of the studied material was represented by clasts from remaniés localized in Brazilian South Shelf (PSB). In this way, the present study also investigated whether modern environmental factors can be deduced from taphonomic profiles of reworked death assemblages. The results here obtained show that, in the PSB, (i) despite absolute differences, the taphonomic profile of micro and macroclasts does not vary, and that (ii) when the study is conducted in larger spatial scales, up to 60% of variation in the taphonomic profile can be explained by the environmental variation. Still in this region, the main environmental factors influencing the damage in bivalve microclasts are: i) salinity, which regards the freshwater input (and also the oceanic circulation as a whole), and ii) the proportion of carbonate in the sediment, which reflects, primarily, the existence of bioclastic deposits (constituted mostly by reworked molluscan shells, or biological remaniés). The proximal bioclastic deposits (south of Lagoa dos Patos outflow) presented the higher evidence of dissolution and physical reworking, while the distal bioclastic deposits (located within the batimetric range of 100 and 200 m) showed better taphonomic preservation. Beyond this, it was also possible to determine that color alteration in bioclasts from PSB (a ubiquitous feature in samples concentrated with gravel and carbonate) is significantly linked to sediment redox status. The color alteration, by turn, are related to the presence of metallic elements (such as iron and manganese), which are important in the biological carbon mineralization, and in the TAZ geochemistry in marine environment. The analysis conducted at geographic spatial scale, including material from Northeastern Brazilian Shelf allowed determining that depth, a significant environmental factor influencing taphonomic damage in PSB, lost importance in large scale. In different geographic provinces, the main factors influencing taphonomic signatures are linked to current velocity, salinity and sedimentary carbonate. These environmental factors reflect the influence of differing water masses (North Brazilian Current and Malvinas Current) and different sedimentary regimes (relict siliciclastics and modern carbonatic).

Made available in DSpace on 2014-10-09T12:35:32Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:04:01Z (GMT). No. of bitstreams: 0 / Moluscos têm sido empregados como bioindicadores em estudos de contaminação ambiental. Nesse contexto, o caramujo de água doce Biomphalaria glabrata tem sido avaliado como um bom modelo laboratorial, e estudos prévios apontaram sua aplicação na pesquisa ambiental. A proteína HSP70 é uma molécula de 70 kDa, pertencente a uma família de proteínas com papel na manutenção da homeostase dos seres vivos: as proteínas de choque térmico (HSPs); e vem sendo estudada como potencial biomarcador de dano ambiental, indicando estresse e protegendo os organismos dos danos às proteínas. Neste trabalho, foi caracterizada a proteína HSP70 de B. glabrata pelo Western blot, com o objetivo de seu emprego em aplicações ambientais futuras. Para isso, caramujos de 5-6 meses de idade, com diâmetro de concha de 14,4 (±1,7) mm, foram expostos ao calor e ao cloreto de cádmio (CdCl2) a fim de se verificar a resposta desta proteína frente a esses estresses. Os animais foram dissecados para investigação da indução da HSP70. As proteínas foram extraídas dos tecidos com tampão RIPA, separadas em eletroforese desnaturante em gel de poliacrilamida, transferidas para uma membrana de nitrocelulose e detectadas com anticorpo específico para HSP70. A CL50/96h foi determinada como sendo 0,34 (0,30-0,37) ppm para o CdCl2 e serviu de referência para os experimentos de indução da proteína. Foi observado que a exposição a temperaturas subletais aumentou a resistência dos caramujos à temperatura letal de 42 °C. Exposições prévias ao calor de 33 °C e ao CdCl2 a 0,22 ppm aumentou a sobrevivência dos caramujos B. glabrata à concentração letal de CdCl2 (0,7 ppm) e à temperatura letal (42 °C), respectivamente. Os achados do Western blot apontaram para um possível papel da HSP70 nesse processo. Os resultados mostraram relação entre a proteína HSP70 e o aumento na sobrevivência aos estímulos letais após prévia exposição a estresses moderados. O Western blot mostrou uma indução da HSP70 nos grupos pré-expostos, se comparados aos grupos controles. A glândula digestiva foi o tecido mais responsivo, no que concerne à indução da proteína HSP70, comparando com tecidos de cabeça/pé e ovoteste. Foi encontrado o pico de indução da HSP70 nos caramujos B. glabrata após 48 horas de exposição ao calor de 33 °C, e após 96 horas de exposição ao CdCl2 a 0,22 ppm. Apesar do bem conhecido papel da HSP70 na termotolerância e tolerância a outros agentes estressores nos organismos vivos, esta foi a primeira vez que isto foi demonstrado no B. glabrata, oferecendo subsídios para a sua aplicação em estudos de monitoramento ambiental. Os resultados apresentados aqui abrem o caminho para estudos futuros dessa proteína no molusco, e fornecem mais bases para o conhecimento do B. glabrata. / Tese (Doutoramento) / IPEN/T / Instituto de Pesquisas Energeticas e Nucleares - IPEN-CNEN/SP

Genetically modified (GM) crops were introduced in South Africa in 1989 and commercially available by 1998. Legislation to control the use of GM crops was only implemented in 1999, with the genetically modified organisms (GMO) act (15 of 1999). In 2012 2.9 million ha of GM crops were planted in South Africa alone. GM Crops, such as Bt maize, are promoted as safer for the environment since no chemical pesticides are needed. However, recently GM crops have been making headlines as more and more studies find adverse effects of these crops on non-target organisms. The effects on aquatic environments have not yet been fully determined, even though traces of Bt residue have been found in water systems surrounding agricultural lands. The aim of this study was to establish the effects of the Bt toxin on fecundity, development and growth of Bulinus tropicus, a freshwater snail. The experiment made use of a static renewal tests to expose B. tropicus to 50 cm2 Bt maize and cotton leaves in 900 ml of synthetic freshwater. The snails were exposed for the duration of one full life cycle (embryo to adult). Endpoints measured included the development, growth, fecundity, and deformities of the reproductive organs. The results obtained showed retarded development and low embryo survival when the snails were exposed to cotton leaves, irrespective of the presence or absence of Bt, indicating to the possibility of trace residues of chemical pesticides may have been present on the leaves. Initial stimulated growth of hatchlings was observed for both Bt cotton and maize exposures, but after sexual maturity has been reached, ‘surplus’ energy was probably shared between growth and fecundity, resulting in a reduction of growth rate. Energy is gained from their diet, thus a sub-optimal diet would result in less energy available to functions such as growth and fecundity. Signs of developmental instability were found in the formation of the shell opening of the snails exposed to Bt. Fecundity decreased significantly after snails had been exposed to Bt maize / cotton leaves. No differences were found in the penis sheath-preputium length ratio, indicating that Bt had no deleterious effects on the reproductive organs. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Genetically modified (GM) crops were introduced in South Africa in 1989 and commercially available by 1998. Legislation to control the use of GM crops was only implemented in 1999, with the genetically modified organisms (GMO) act (15 of 1999). In 2012 2.9 million ha of GM crops were planted in South Africa alone. GM Crops, such as Bt maize, are promoted as safer for the environment since no chemical pesticides are needed. However, recently GM crops have been making headlines as more and more studies find adverse effects of these crops on non-target organisms. The effects on aquatic environments have not yet been fully determined, even though traces of Bt residue have been found in water systems surrounding agricultural lands. The aim of this study was to establish the effects of the Bt toxin on fecundity, development and growth of Bulinus tropicus, a freshwater snail. The experiment made use of a static renewal tests to expose B. tropicus to 50 cm2 Bt maize and cotton leaves in 900 ml of synthetic freshwater. The snails were exposed for the duration of one full life cycle (embryo to adult). Endpoints measured included the development, growth, fecundity, and deformities of the reproductive organs. The results obtained showed retarded development and low embryo survival when the snails were exposed to cotton leaves, irrespective of the presence or absence of Bt, indicating to the possibility of trace residues of chemical pesticides may have been present on the leaves. Initial stimulated growth of hatchlings was observed for both Bt cotton and maize exposures, but after sexual maturity has been reached, ‘surplus’ energy was probably shared between growth and fecundity, resulting in a reduction of growth rate. Energy is gained from their diet, thus a sub-optimal diet would result in less energy available to functions such as growth and fecundity. Signs of developmental instability were found in the formation of the shell opening of the snails exposed to Bt. Fecundity decreased significantly after snails had been exposed to Bt maize / cotton leaves. No differences were found in the penis sheath-preputium length ratio, indicating that Bt had no deleterious effects on the reproductive organs. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Molluscan succession was studied in 18 abandoned limestone quarries of different age in Bohemian Karst. The number of species in studied quarries increases in the time. The less species were found in the oldest quarries. This trend was probably caused by habitat diversity decline although it is not conclusive. The best predictors of mollusk species composition are type of surrounding vegetation, light and the cover of tree layer. It is important to take into account correlations of the other environmental variables with light and the cover of tree layer. Compositions of mollusk assemblages of north and south-facing slopes differ, but these differences diminish with ongoing succession. Quarries represent suitable model sites for the study of succession.

O atual conhecimento sobre a margem do manto em moluscos bivalves é extenso, incluindo informações sobre morfologia, função e diversidade. Bivalves da família Pectinidae, também conhecidos como vieiras, possuem complexa margem palial, organizada em três pregas, incluindo olhos e tentáculos. Questões acerca do desenvolvimento da margem do manto em bivalves continuam amplamente incompreendidas, assim como a relação entre características paliais ao longo dos diferentes estádios do ciclo de vida. Neste contexto, a presente investigação utilizou a espécie de vieira Nodipecten nodosus como modelo para compreensão da morfogênese da margem palial em Pectinidae, com ênfase na origem e diferenciação das pregas paliais e estruturas associadas. Para contemplar esses objetivos, espécimes em diferentes estádios de desenvolvimento larval e pós-metamórfico foram analisados por meio de técnicas integradas de microscopia (i.e., histologia, microscopia eletrônica de varredura e transmissão, e imunocitoquímica aplicada à microscopia confocal). Inicialmente, a margem palial em larvas véliger de N. nodosus não é pregueada, porém, ao longo do desenvolvimento, dois processos de evaginação são determinantes na formação das pregas paliais. O primeiro ocorre no estádio de pedivéliger, originando as pregas externa e interna, bem como o sulco do perióstraco. O segundo ocorre após a metamorfose, sendo responsável pela origem da prega palial mediana a partir da porção interna da prega interna. Os sistemas muscular e nervoso da margem palial têm origem durante o período larval, tornando-se amplamente desenvolvidos posteriormente. Estruturas associadas, como tentáculos e olhos paliais, são formadas apenas após a metamorfose, e compõem a complexa condição final da margem do manto em Pectinidae. Os diferentes tipos tentaculares possuem desenvolvimento e anatomia similar, entretanto diferem quanto ao tamanho, tipo de musculatura, organização ciliar e presença de células glandulares. Os olhos paliais em formação diferenciam-se gradualmente em sentido proximal-distal, essas características morfológicas sugerindo um nível simples de fotopercepção direcional como condição inicial. Os dados aqui apresentados para N. nodosus permitiram propor um modelo geral para o desenvolvimento da margem palial em Pectinidae, além de contribuir para o entendimento da morfogênese dessa região em Bivalvia / Current knowledge of the bivalve mantle margin is extensive, covering several aspects of its morphology, function and diversity. Bivalves from the family Pectinidae, also known as scallops, bear three pallial folds at the mantle margin, including complex structures, such as tentacles and eyes. The development of the bivalve mantle margin is still poorly understood, the morphogenesis and functional anatomy of mantle margin features during developmental stages being enigmatic. The present investigation used the scallop Nodipecten nodosus (L. 1758) as a model species to understand mantle margin morphogenesis in the Pectinidae, with emphasis on the origin and differentiation of pallial folds and associated pallial structures. To achieve these goals, specimens from larval and postmetamorphic stages were thoroughly analyzed by means of integrative microscopy techniques (i.e., histology, scanning and transmission electron microcopy, and immunocytochemistry combined with confocal microscopy). In veliger larvae of N. nodosus, the mantle margin is initially unfolded, two folding processes being crucial for pallial fold establishment during further development. The first one occurs by the pediveliger stage, forming the outer and inner folds, as well as the periostracal groove. The second folding process takes place after metamorphosis and is responsible for the formation of the middle pallial fold from the inner region of the inner mantle fold. The emergence of muscular and nervous systems in the mantle margin occurs early during development, at the larval stage. Associated pallial structures, including tentacles and eyes, develop only after metamorphosis, and contribute to the complex final condition of the mantle margin in Pectinidae. Although different tentacular types have similar development and anatomy, they differ in size, muscle type, ciliary organization, and gland cells distribution. Developing pallial eyes exhibit gradual differentiation in a proximal-distal direction, and their morphological features suggest a simple level of directional photoreception as the initial ocular condition in juveniles. The present investigation conducted with N. nodosus provided a general model to understand mantle margin development in the Pectinidae, as well as insights into the morphogenesis of this region in the Bivalvia

A Actuopaleontologia é o ramo da Paleontologia que visa aperfeiçoar as interpretações paleoambientais e paleoecológicas através das assinaturas tafonômicas, principalmente aquelas presentes em conchas bivalves. Tais estudos tafonômicos com bivalves marinhos têm sido conduzidos em ambientes sedimentares do Holoceno, porém, no Brasil, investigações similares são ainda incipientes, tendo sua predominância no litoral norte do Estado de São Paulo. O Complexo Estuarino de Paranaguá (CEP), situado na porção centro-norte do litoral do Paraná, possui feições fisiográficas distintas das encontradas no litoral norte do Estado de São Paulo. Com o objetivo de gerar dados comparativos entre os diferentes ambientes da costa brasileira, este estudo pretende caracterizar tafonomicamente as associações de moluscos bivalves em 11 pontos de coleta ao longo do CEP. Foram coletados oito litros de sedimento com a utilização de pegador de fundo tipo Petersen, em parceria com o Laboratório de Oceanografia Geológica (LOGeo CEM/UFPR). As assinaturas tafonômicas analisadas, podendo ser de origem química, física e/ou biológica, foram: articulação, tipo de valva, fragmentação, modificação da margem da concha, corrasion, perióstraco, brilho, cor, bioerosão, incrustação e predação. A ação dessas assinaturas pode resultar em outra assinatura tafonômica aqui analisada, a alteração da textura da superficial da concha. Foram analisados 150 bioclastos de cada um dos pontos de coleta, e cada assinatura tafonômica foi classificada individualmente. Os bioclastos foram peneirados em malhas de 2 mm para que as menores classes de tamanho não fossem excluídas das análises tafonômicas. Além dos bioclastos, foram coletadas amostras da água junto ao fundo dos locais amostrados, para registro de pH, salinidade, temperatura e turbidez. Ao total, 1.438 bioclastos foram utilizados nas análises tafonômicas e, das 11 estações de coleta, apenas duas não apresentaram o n mínimo de 150 conchas. Os dados abióticos não apresentaram grandes diferenças nas diferentes estações de coleta. A análise de cluster foi feita levando-se em consideração todas as assinaturas tafonômicas estudadas, e um dendograma geral relacionou os 9 pontos de coleta em que obteve-se o mínimo de 150 bioclastos. O dendograma gerado nesta análise mostrou quais as acumulações de moluscos bivalves e seus respectivos pontos de coleta estão mais semelhantes. Há um primeiro grupo formado pelas estações 683, 336, 482, 696 e 32. Com exceção da estação 32, as estações reunidas no primeiro grupo são os pontos de coleta mais próximos às desembocaduras do CEP, região de maiores profundidades e fortes correntes de marés. O segundo grupo é formado pelas estações 607, 152, 645 e 472, sendo pontos de coleta mais internos na área do CEP, próximos a ilhas, manguezais e bancos de areia. As assinaturas alteração da cor, brilho, modificação da margem, perióstraco e corrasion não parecem ser assinaturas úteis na identificação ou para uma possível caracterização dos diferentes ambientes do CEP. Isto porque elas não são variáveis entre esses diferentes ambientes. Houve predominância de desarticulação e fragmentação das valvas em todos os ambientes estudados, sendo possível que a fragmentação esteja sobre maior influência dos danos biológicos. Bioerosão e incrustação não apareceram em frequências expressivas, porém, apenas as estações de coleta próximas a desembocadura do CEP apresentaram bioeorosão, enquanto incrustação apareceu em todas as estações. A textura superficial da concha correlacionou-se positivamente com as assinaturas corrasion e bioerosão. / Actuopaleontology is the branch of Paleontology that aims to enhance paleoecological and paleoenvironmental interpretations, through the taphonomic signatures, especially those in bivalve shells. Such taphonomic studies with marine bivalves have been conducted in sedimentary Holocene environments, however, in Brazil, similar investigations are still incipient, having its predominance in the northern coast of São Paulo. The Paranaguá Estuarine Complex (CEP), situated in the north central portion of the Paraná coast, has different physiographic characteristics of those found in the north coast of São Paulo. In order to generate comparative data between different environments of the Brazilian coast, this study aims to characterize the taphonomy of bivalve mollusk associations from 11 collection stations along the CEP. Eight liters of sediment were collected with background catcher Petersen, in partnership with the Laboratory of Geological Oceanography (LOGeo - CEM/UFPR). The taphonomic signatures analyzed, which may be of chemical, physical and/or biological origin, were: articulation, valve type, fragmentation, edge modification, corrasion, periostracum, brightness, color, bioerosion, encrustation and predation. The action of these signatures may result in other taphonomic signature analyzed here, the alteration of shell superficial texture. 150 bioclasts were analyzed from each collection point and each taphonomic signature was individually classified. The bioclasts were sieved through mesh of two millimeter, so that classes of smaller size were not excluded from the taphonomic analyzes. In addition, water samples were collected at the bottom of each station for analysis of pH, salinity, temperature and turbidity. In total, 1,438 bioclasts were used in the taphonomic analysis and only two stations did not show the minimum n of 150 shells. Abiotic data showed no great differences in the different sampling stations. The cluster analysis was performed taking into account all studied taphonomic signatures, and a general dendrogram listed the collection points that showed the minimum of 150 bioclasts. The dendrogram generated in this analysis showed which accumulations of bivalve molluscs and their respective collection points are more similar. There is a first group formed by the stations 683, 336, 482, 696 and 32. With the exception of station 32, the stations on the first group are the closest to the mouths of the CEP, region of greater depths and strong tidal currents. The second group is formed by stations 607, 152, 645 and 472, being the more internal collection points, near islands, mangroves and sandbanks. The signatures color, brightness, edge modification, periostracum and corrasion do not seem to be useful in identifying or for a possible characterization of the different environments of the CEP, because they are not variable between these different environments. There was predominance of disarticulation and fragmentation of the valves in all study sites, being possible that the fragmentation was influenced more by biological damages. Bioerosion and encrustation did not appear in expressive frequencies, however, only the stations near the mouth of the CEP presented bioeorosão, while encrustation appeared in all stations. The surface texture of the shell was correlated positively with the sigantures corrasion and bioerosion.

Zinc, copper and cadmium accumulation was measured in two herbivorous gastropod molluscs Austrocochlea constricta and Bembicium auratum from Lake Macquarie NSW an area with a history of trace metal pollution. The investigation consisted of three main parts. The first part examined the influence of organism mass and location within the Lake on whole body tissue metal concentrations. This part of the study also compared the distributions of tissue metal concentrations of populations from Lake Macquarie, a known polluted environment, with those of populations from Jervis Bay NSW, an unpolluted environment, to establish whether either species is a net accumulator of zinc copper or cadmium. The second part of the investigation examined a range of factors which may influence whole body metal concentrations. One location in Lake Macquarie was sampled monthly from August 1995 to July 1996. The factors examined were temporal variation, gender, breeding cycle, and tissue distribution. The final part of the investigation examined the detoxification and storage of excess metals in the gastropods from Lake Macquarie. The mechanisms studied were metallothioneins and granules. The tissue metal concentrations of both species were found to be independent of mass. Location within Lake Macquarie did not significantly influence tissue metal concentrations. Variation between individuals was the most significant contribution to overall variation, resulting in a positive skewing of sample trace metal distributions. B. auratum populations from Lake Macquarie had significantly higher copper and cadmium tissue concentrations and A. constricta populations had significantly higher zinc, copper and cadmium tissue concentrations than the populations from Jervis Bay. This suggests that regulation of these metals is not occurring. A. constricta may therefore be considered a net accumulator of zinc, copper and cadmium and B. auratum of copper and cadmium. Tissue metal concentrations did not vary significantly over time. It is suggested that the organisms are in equilibrium with their environment. B. auratum has higher natural equilibrium concentrations than A. constricta particularly for copper and cadmium, suggesting different routes of exposure, uptake or accumulation for the two species. Gender and breeding cycle did not significantly influence tissue metal concentrations. Most of the variability in total copper and cadmium concentrations of both species was explained by variability in gonad tissue metal concentration, while variability in the gonad and somatic tissues zinc concentration explained about an equal amount of the variability in total zinc concentration. A. constricta and B. auratum were both found to induce a cadmium binding protein which has some features in common with metallothionein. A protein of around 10 000 Da which binds approximately 60% of the soluble cadmium was isolated using gel filtration. This protein was further separated into two isoforms using anion exchange. The first isoform eluted at the same time as MT I and the second at the same time as MT II rabbit liver standard. Large cells containing granular material which stained positive for calcium were observed interspersed among the connective tissue immediately behind the columnar epithelial cells lining the gut wall in both species under a light microscope. Calcium positive granular particles were also observed within the columnar epithelial cells of B. auratum. These species have been shown to be net accumulators of the trace metals investigated, with the exception of zinc in B. auratum. It has also been established that organism mass, gender and reproductive state, the partitioning of metals between tissues, and temporal effects are not confounding factors for the purposes of comparing trace metal concentrations between populations. They should therefore be effective biomonitors of the trace metals investigated, with the exception of zinc in B. auratum.

Vibrio spp., natural inhabitants of aquatic environments, are one of the most common causes of bacterial gastroenteritis in the world, being spread to humans via the ingestion of seafood, contaminated drinking water or exposure to seawater. The majority of Vibrio spp. are avirulent, but certain strains may sporadically be human pathogenic. Vibrio cholerae may cause cholera and fatal wound infections, Vibrio parahaemolyticus may cause gastroenteritis and Vibrio vulnificus may cause wound infections and sepsis. To expand current knowledge of the occurrence, ecological niche and persistence of potential human pathogenic Vibrio spp. in aquatic environments, occurrence and laboratory studies were performed. The seasonal variation of Vibrio spp. in clams and mussels from Mozambique and Sweden were studied, with isolated strains characterized and compared with those isolated from water samples collected in India. Results showed that the numbers of Vibrio spp. in Mozambican clams peaked during the warmer rainy season and that the dominating species was V. parahaemolyticus. Biochemical fingerprinting and virulence screened by PCR revealed a high similarity among strains from the different aquatic environments. However, isolate functional hemolytic analyses and antibiotic resistance patterns differed between strains; Swedish and Indian strains were less sensitive to the tested antibiotics and had a lower hemolytic capacity than those from Mozambique. Molecular analysis of bacterial DNA from Swedish mussels showed the presence of the three Vibrio spp. most commonly linked with human illness, as well as their associated virulence genes. The strains isolated from marine and clinical environments were equally and highly harmful to the tested eukaryotic cells. The persistence of clinical V. cholerae in aquatic environments was investigated in vivo. Strains were exposed to mussels, with bacterial uptake and elimination then examined. The mussels were able to avoid the most potent strain by complete closure of shells. The less potent strain was accumulated in mussel tissue in low levels and one marine control strain to a higher degree. Mussels eliminated the pathogenic strain less efficiently than they did the marine strain. One clinical and one marine strain were then exposed to 4°C for 21 days, with the temperature then increased to 20°C. The clinical strain was more prone to lose culturability than the marine strain at 4°C, the former performed significantly better in regaining culturability after the temperature up-shift. Subsequently, the persistence of the clinical strain in natural bottom sediment, incubating as above, was studied and results showed a similar decrease in culturable numbers in the sediment as in the water. As the clinical V. cholerae strains did not carry any of the standard set of virulence genes, the ability to change from non-culturable to culturable may be of great importance to strain pathogenicity. The results also show that natural bottom sediment may be a potential reservoir of human pathogenic Vibrio spp.

The ecological impact of past extinction events is one of the central issues in paleobiology. In face of present environmental changes, a better understanding of past extinctions will enable us to identify the magnitude of biodiversity crises and their underlying processes. The Late Neogene was a time of extraordinary climatic reorganization, including Northern Hemisphere glaciation, the rise of the Central American Isthmus, and associated changes in environmental conditions. Therefore, the Late Neogene extinctions of marine molluscs of South Florida present an ideal platform to examine the interaction between environmental changes and biotic response. In the present study, three different aspects of the Late Neogene extinctions are examined: temporal diversity patterns, selectivity patterns, and the impacts of these extinction events on ecological interactions. In the first part of this study, the diversity pattern of marine bivalves of Florida during the Late Neogene. Using bulk samples enables to take account of varying sampling intensity and underlying relative abundance distributions in diversity estimation. Comparison of sample-standardized diversity analyses shows that both richness and evenness of marine bivalve community declined at the Tamiami - Caloosahatchee transition, which coincides with the proposed first phase of the Late Neogene extinctions at the end of the Pliocene. Although magnitude of biodiversity loss was severe during these late Neogene extinction events, extinction risk was non-randomly distributed across taxa. Selectivity analyses, a combination of both commonly used non-parametric tests and logistic regressions, suggest that abundance or local population size was positively related with survivorship during the late Neogene. As other biological or ecological traits can influence this observed relation between abundance and extinction vulnerability, multivariate approach is used to control for these traits. Even after effects of geographic range and feeding mode is considered, the positive relation between abundance and survivorship, which supports predictions from biological studies, is evident in case of these Late Neogene extinction events. While present analyses show that the increase in relative abundance of Chione is a major factor in driving changes in community compositions, interactions between Chione and its' drilling predators also varied during the Late Neogene. This study suggests that identification of predators is a critical part of evaluation of prey-predator interactions. When drill hole traces of two predatory gastropod groups, muricids and naticids, are differentiated based on a revised site selectivity criteria, temporal trend of prey size selectivity differs from previous reports. Both groups exhibit some changes in predatory behavior during phases of the Late Neogene extinctions, suggesting that previous hypothesis of prey turnover at the Caloosahatchee - Bermont transition cannot explain the observed temporal trends of prey size selectivity in the present study.