Many studies addressing the toxicity of silver nanomaterials have found that smaller sized silver nanoparticles are usually more toxic to organisms and in cell culture than particles of larger sizes yet it is not entirely clear why. We investigated the size dependent toxicity of silver nanoparticles by measuring the response of embryonic zebrafish (Danio rerio) following exposure to a library of thirteen distinct silver nanoparticle size distributions with mean diameters between 8.9 nm and 112.6 nm. Data analysis using dose���response modeling revealed that silver nanoparticles (AgNP) induced embryo toxicity that is dependent on the total surface area and not on the mass or particle number in solution. Included in this study is a comparison between embryo toxicity induced by silver nitrate (AgNO���) and AgNPs for cardiovascular endpoints, as well as an investigation into the influence of the chorion on AgNP toxicity. This study demonstrates the importance of using alternative dose metrics in nanotoxicology, and highlights the value of using the embryonic zebrafish to explore nanomaterial structure activity relationships. / Graduation date: 2013

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

University of Macau / Institute of Chinese Medical Sciences

Risk assessment of environmental exposure to chronic, trace concentrations of contaminants presents an analytical challenge to interpret data in a biologically meaningful way. Biomarkers are compounds that can provide integrated information concerning the effects of contaminants on biochemical processes. The metal-binding protein, metallothionein (MT), is a biomarker of toxic heavy metals, such as cadmium (Cd), since these metals bind to MT in vivo and induce transcription of MT genes with subsequent MT protein synthesis. A high performance liquid chromatography (HPLC)-thermospray microatomization-atomic absorption spectroscopy (AAS) method was developed for detecting Cd proteins from two invertebrate models; freshwater mussels (Pyganodon grandis) from a whole lake ecosystem exposure to Cd (Experimental Lakes Area, Canada), and zebra mussels (Dreissena polymorpha) recently introduced to North America. Methods for coupling gel filtration HPLC to AAS or inductively coupled-mass spectrometry (ICP-MS) were developed to provide sensitive and selective information on metal-binding proteins in freshwater mussel extracts. Sensitive metal detection by these methods necessitated precautions to minimize HPLC column-protein exchange of metals. These interactions were relevant to the determination of low concentrations of mussel MT-like proteins (MLPs) by these metal-specific detection systems. Saturation methods employing Cd as a metallic marker for the quantification of characterized MTs were adapted for freshwater mussels exposed to low environmental Cd concentrations. Characterization of Cd-saturated mussel extracts by HPLC-ICP-MS demonstrated the presence of copper and zinc, metals physiologically bound to MT, in the principal metal-binding fraction of mussel extracts. Experimental results also indicated that mussel MLPs are not as heat-stable as mammalian MT. Zebra mussels were chosen as a bioindicator species for obtaining a standard of freshwater MT biomarker for toxi

Découvert en 1964, le virus d'Epstein-Barr appartient à la famille des gamma-herpèsvirus. Ce virus à ADN présente une forte prévalence (90% de la population adulte est infectée). Ce fut le premier virus identifié comme associé à des cancers (lymphome de Burkitt et d'Hodgkin, carcinome gastrique et de l'oropharynx). Ce virus a pour spécificité de posséder deux cycles distincts : latent et lytique (production de particules virales). Le facteur de transcription viral EB1 (ou Zebra) est un élément clé lors de l'initiation du cycle lytique et semble une cible importante pour l'élaboration de nouveaux traitements. Une première partie de ce travail concerne la caractérisation d'une protéine cellulaire (l'Ubinucléine) qui interagit et inhibe l'activité de EB1. Cette protéine voyage entre noyaux et jonctions serrées. Elle appartient à la famille des " NACos " (nuclear and adhesion complex components). La fonction de l'Ubinucléine n'est pas connue et sa protéomique quand elle est localisée dans les jonctions serrées, a été réalisée. Des études fonctionnelles montrent que l'Ubinucléine interagit avec plusieurs partenaires cellulaires, emprunte la voie d'endocytose dépendante de la clathrine et que sa localisation cellulaire (nucléaire ou dans les jonctions serrées) est affectée par la PKA. Dans une seconde partie, nous nous sommes intéressés à des molécules inhibitrices du facteur de transcription viral EB1. Après criblage à haut débit de composés chimiques (EMBL-Heidelberg), des tests in-vitro ont permis d'en sélectionner un pour des essais in-vivo. Ce composé chimique inhibe l'activité du facteur de transcription EB1 et bloque précocement la mise en place du cycle lytique dans des cellules de lymphome de Burkitt. Il semble donc intéressant d'améliorer l'efficacité et la spécificité de cette molécule.

Zooplankton samples collected May through August, 1990 to 1996 at site M, 15 m depth in the Indiana waters of Lake Michigan were examined for microcrustaceans, rotifers, and zebra mussel (Dreissena polymorpha) veligers. Abundance numbers per m3 were combined with Coffman's (1995) data to quantify recent trends in the zooplankton.Rotifers dominated the community, with Polyarthra vulgaris, Colurella, Keratella crassa, and K. cochlearis the most prevalent species. Copepods, primarily nauplii and copepodid forms, were the next most abundant taxa. Except for 1994 and 1995, Cladocera abundance was higher than zebra mussel veliger abundance. The cladoceran assemblage was dominated by Bosmina Iongirostris, with very few Daphnia noted. A precipitous decrease in zebra mussels veligers from 1994 to 1996 warrants further investigation to determine the nature and the extent of the population decline.Regression analysis of zebra mussel veliger abundance against abiotic and biotic factors demonstrated veligers varied directly with rotifer abundance and water column temperature and indirectly with cladoceran and yellow perch (Perca flavescens) abundance. A similar regression model of rotifer abundance revealed direct correlation with calanoid, cladoceran, zebra mussel veliger, and yellow perch abundance. / Department of Biology

Biological invasions threaten the stability and biodiversity of freshwater ecosystems worldwide. The impacts of an invading species often vary across systems, making their prediction difficult. When data from multiple invaded sites are available, statistical models can be developed to correlate an invader's distribution and abundance with local environmental variables; such models could then provide managers with useful tools to help prioritize efforts to control the invader. The introduction of the zebra mussel (Dreissena polymorpha) and quagga mussel (D. bugensis) to North America ranks among the most ecologically and economically disruptive aquatic invasions ever documented. While some attempts have been made to predict zebra mussel occurrence and abundance, none have been made for quagga mussels. Furthermore, few studies have been based on river systems, which possess the bulk of North American freshwater biodiversity. I related zebra and quagga mussel occurrence and biomass to physical habitat variables (calcium concentration, substrate size and depth) in the St. Lawrence River. I then developed predictive models of abundance for each species from combinations of these variables. Each variable explained a significant amount of variation in mussel biomass, but different combinations of variables were obtained for each species. Although these models do not account for all of the variation in abundance, they do provide a useful basis for predicting dreissenid distribution and abundance in other invaded river systems.

Biological invasions are a global phenomenon that can threaten native species and disrupt ecosystem processes. Exotic species also impact ecosystems in less conspicuous ways by provoking native species to alter their foraging behaviour. Subtle impacts such as diet shifts are frequent, and can have consequences for food web dynamics and the fitness of native predators. Diet shifts involving the consumption of exotic species require native predators to recognize, capture and handle novel prey. In this thesis, I document a diet shift in the St. Lawrence River involving a common native fish and Eurasian mussels that invaded the river in the early 1990s. I conducted diet analysis of yellow perch (Perca flavescens) at multiple sites in the upper St. Lawrence River and discovered that they consumed substantial quantities of zebra and quagga mussels (Dreissena spp.) in the Soulanges Canal, an artificial waterway west of Montreal. This was unexpected because perch lack adaptations for crushing molluscs. This foraging innovation was not observed at the same site in the early 1990s or at other sites at any time. Mussel shells were weaker at this site, probably because of exposure to calcium-poor water. This suggests that water chemistry mediates yellow perch predation on mussels. This study provides an example of diet shifts involving exotic prey and illustrates the influence of abiotic factors on species interactions.

ABSTRACT The zebra shark Stegostoma fasciatum is an oviparous, demersal carpet shark that forms temporary aggregations and is distributed within shallow, coastal, subtropical and tropical waters of the Indo-West Pacific (IWP) region. The IWP region encompasses various densities of human habitation leading to differences in the levels of exploitation of chondrichthyan fishes, which are targeted for local consumption and export. This is reflected in the two current regional IUCN Red List classifications for zebra sharks of ‘Least Concern’ and ‘Vulnerable’. Despite the conservation concerns for zebra sharks in the vulnerable parts of their range, as well as importance within the ecotourism industry and collection for aquarium trade, little is known about this species in the wild. The aim of this thesis was to conduct the first intensive examination of population ecology in the zebra shark by: (i) investigating the population structure and environmental parameters associated with a relatively unexploited aggregation of zebra sharks in south-east Queensland, Australia; and (ii) investigating the genetic continuity between zebra shark populations in areas with differing levels of exploitation. To investigate the size and structure of the aggregation at the site The Group in south-east Queensland, photo-identification and mark-recapture methods were employed over a three year period (2003 - 2006). In total 327 individual zebra sharks were identified from 570 photographs. Numbered dart tags on 15 zebra sharks were used to confirm pigmentation patterns were unique and persistent in wild zebra sharks for up to 810 days. Pollock’s robust design resulted in an annual population estimate of 458 individuals (95% CI = 298 – 618). The mean number of zebra sharks observed on a single day was 8 (± 8 SE) and the maximum number of zebra sharks seen on a single day was 34. In total, 27% of the sharks were sighted in more than one summer aggregation period and males had greater re-capture probabilities than females. The aggregation consisted exclusively of large (>1800 mm total length) adults with an overall female sex bias of 3.8:1, though sex-ratios varied temporally. Passive acoustic telemetry techniques were employed to investigate the seasonality and residency of the sharks and the environmental parameters associated with their presence at the study site. Five VR2 acoustic receivers were deployed for a period of 21 months (December 2004 – August 2006) and recorded ten acoustically-tagged zebra sharks (seven females and three males). The sharks showed clear seasonality in their visitation patterns with greatest presence during the austral summer months of November through to February and no presence at all during the winter months from June through to October. However their presence did not coincide with peak water temperatures. Rather there appeared to be a critical minimum water temperature of approximately 22 ºC, above which zebra sharks would be present. The tagged zebra sharks displayed greater visitation during the calmest sea conditions. The sharks further displayed diel periodicity with greater during daylight hours, and may be using The Group as a resting site during the inactive diurnal period of the day. Complementing the ecological study on the aggregation, the population genetic structure of 180 zebra sharks from 13 locations throughout the IWP was investigated to test the concordance of IUCN zones with demographic units that have conservation value. A zebra shark specific microsatellite library was developed consisting of 14 polymorphic loci of which 13 were employed for the population genetic analysis along with mitochondrial ND4 sequence data. Mitochondrial and microsatellite data-sets from samples collected throughout northern Australia and south-east Asia concord with the regional IUCN classifications. However, evidence of genetic subdivision was found within these regions, including subdivision between locations connected by habitat suitable for migration. Further, parametric FST analyses and Bayesian clustering analyses indicated the primary genetic break within the IWP is not represented by the IUCN classifications but rather is congruent with the Indonesian throughflow current. This study has demonstrated that zebra sharks form a large, predictable, shallow coastal aggregation comprised entirely of mature adults as well as display high genetic subdivision in the Indo-West Pacific region with little to no recruitment into over-exploited populations from healthy populations. These findings indicate that, if zebra sharks form similar aggregations as that which occurs at The Group in regions of high fishing pressure then, without sufficient protection and management localised depletion and extirpation will eventuate for zebra shark populations in heavily-fished areas.