Philosophiae Doctor - PhD / In the marine environment, penguins have been described as curators and serve a critical role in ecological balance. The African penguin (Spheniscus demersus) has undergone a rapid population decline, mainly due to disturbances in their natural habitat. The African penguin was up-listed from vulnerable to endangered on the IUCN Red List for Threatened Species in 2010 and thus urgent conservation action is required. Integral to long-term conservation action of any species is a basic knowledge of its reproductive biology, which is currently lacking for African penguins. The main aim of this investigation was to evaluate techniques for the collection of semen in African penguin and to determine sperm quality in order to cryopreserve sperm for in vitro fertilization (IVF) purposes of captive and wild populations. Semen was collected once a week during two breeding seasons from two captive African penguins. Ejaculates (n=51) were obtained over two breeding seasons (Jan-Feb and Jun-Oct) and evaluated for semen volume, sperm concentration, sperm vitality, sperm motility and sperm morphology. In addition twelve (six females and six males, n=4 were breeding pairs) captive African penguins were monitored for hormone (estradiol, testosterone, progesterone) levels prior to and after the egg-laying period.

Ge, Tianfang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 87-93). / Abstract also in Chinese. / EXAMINATION COMMITTEE LIST --- p.ii / DECLARATION --- p.iii / ACKNOWLEDGEMENTS --- p.iv / ABSTRACT --- p.v / ABSTRACT IN CHINESE --- p.vii / ABBREVIATIONS --- p.ix / TABLE OF CONTENTS --- p.xiii / Chapter 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Erythropoiesis --- p.2 / Chapter 1.2 --- The TF-1 model --- p.3 / Chapter 1.3 --- The erythroid marker glycophorin A (GPA) --- p.4 / Chapter 1.4 --- Reactive oxygen species (ROS) --- p.4 / Chapter 1.5 --- Oxidative stress in human erythrocytes --- p.6 / Chapter 1.6 --- Antioxidant defense systems --- p.6 / Chapter 1.7 --- Glucose provides the majority of reducing equivalents in human erythrocytes --- p.9 / Chapter 1.8 --- Glucose transporter type 1 (Glut l) transports glucose and vitamin C into human erythrocytes --- p.10 / Chapter 1.9 --- Hypothesis and objectives --- p.11 / Chapter 1.10 --- Long-term significance --- p.12 / Figure 1.1 Stages of mammalian erythropoiesis. Adapted from (Koury et al.，2002) --- p.13 / "Figure 1.2 Conversion of major ROS. Adapted from (Ghaffari," --- p.14 / Figure 1.3 Major oxidative defense in human erythrocytes --- p.15 / "Figure 1.4 Peroxide scavenging systems. Adapted from (Day," --- p.16 / Chapter 2 --- MATERIALS AND METHODS --- p.17 / Chapter 2.1 --- Cell culture --- p.18 / Chapter 2.1.1 --- Culture media --- p.18 / Chapter 2.1.2 --- Cell maintenance --- p.19 / Chapter 2.1.3 --- Cell cryopreservation --- p.19 / Chapter 2.1.4 --- Cell differentiation --- p.20 / Chapter 2.1.5 --- Cell treatments --- p.20 / Chapter 2.1.5.1 --- Antioxidant treatments --- p.21 / Chapter 2.1.5.2 --- H2O2 challenging --- p.22 / Chapter 2.1.5.3 --- Antibiotic treatment --- p.22 / Chapter 2.2 --- Flow cytometry --- p.23 / Chapter 2.2.1 --- Flow cytometers --- p.23 / Chapter 2.2.2 --- Analysis of erythroid differentiation --- p.23 / Chapter 2.2.3 --- Analysis of cell lineage --- p.24 / Chapter 2.2.4 --- Analysis of intracellular ROS --- p.24 / Chapter 2.2.5 --- Analysis of mitochondrial transmembrane potential (Δψm) --- p.25 / Chapter 2.2.6 --- Analysis of mitochondrial mass --- p.25 / Chapter 2.2.7 --- Analysis of cell death --- p.26 / Chapter 2.2.8 --- Analysis of caspase-3 activity --- p.27 / Chapter 2.2.9 --- FACS cell sorting --- p.27 / Chapter 2.2.10 --- Two-variant flow cytometric experiments --- p.28 / Chapter 2.2.11 --- Analysis of flow cytometry data --- p.28 / Chapter 2.2.12 --- Compensation --- p.29 / Chapter 2.2.12.1 --- Compensation matrix for Annexin V-PI double-staining --- p.29 / Chapter 2.2.12.2 --- Compensation matrix for Annexin V-TMRM double-staining --- p.30 / Chapter 2.2.12.3 --- Compensation matrix for CFSE- GPA double-staining --- p.31 / Chapter 2.2.12.4 --- Compensation matrix for CFSE- TMRM double-staining --- p.31 / Chapter 2.2.12.5 --- Compensation matrix for CM- H2DCFDA-GPA double-staining --- p.32 / Chapter 2.2.12.6 --- Compensation matrix for GPA- TMRM double-staining --- p.33 / Chapter 2.3 --- Western blot --- p.35 / Chapter 2.4 --- Statistical analysis --- p.37 / Chapter 3 --- RESULTS AND DISCUSSION --- p.38 / Chapter 3.1 --- The cells with high GPA staining were younger in cell lineage --- p.39 / Chapter 3.2 --- ROS was produced during TF-1 erythropoiesis --- p.40 / Chapter 3.3 --- ROS production was not essential for TF-1 erythropoiesis --- p.41 / Chapter 3.4 --- ROS production was not the cause of cell proliferation during TF-1 erythropoiesis --- p.41 / Chapter 3.5 --- ROS production was not the cause of sub-lethal mitochondrial depolarization in TF-1 erythropoiesis --- p.42 / Chapter 3.6 --- The cells showing mitochondrial depolarization were mother cells that gave rise to differentiating cells --- p.44 / Chapter 3.7 --- ROS production was not the cause of cell death in TF-1 erythropoiesis --- p.45 / Chapter 3.8 --- ROS production confers oxidative defense during TF-1 erythropoiesis --- p.47 / Chapter 3.8.1 --- Glut l inhibition partially blocked TF-1 erythropoiesis without affecting cell viability --- p.47 / Chapter 3.8.2 --- Antioxidant defense systems were established during TF-1 erythropoiesis --- p.48 / Chapter 3.8.3 --- Antioxidant treatments blocked the establishment of antioxidant defense systems during TF-1 erythropoiesis --- p.51 / Chapter 3.9 --- Conclusion --- p.55 / Chapter 3.10 --- Future work --- p.56 / Figure 3.1 Cell lineage versus erythroid marker during erythropoiesis under vitamin E treatment --- p.59 / Figure 3.2 ROS production during erythropoiesis --- p.60 / Figure 3.3 ROS production versus erythroid marker during erythropoiesis under vitamin E treatment --- p.61 / Figure 3.4 Percentage of ROS+ cells in vitamin E-treated TF-1 erythropoiesis as compared to control --- p.63 / Figure 3.5 Percentage of GPA+ cells in vitamin E-treated TF-1 erythropoiesis as compared to control --- p.64 / Figure 3.6 Cell death versus mitochondrial transmembrane potential (Δψm) during erythropoiesis under vitamin E treatment --- p.65 / Figure 3.7 Erythroid marker versus mitochondrial transmembrane potential (Δψm) during erythropoiesis under vitamin E treatment --- p.67 / Figure 3.8 Cell lineage versus mitochondrial transmembrane potential (Δψm) during erythropoiesis under vitamin E treatment --- p.69 / Figure 3.9 Change of mitochondrial mass during erythropoiesis --- p.71 / Figure 3.10 ROS production versus erythroid marker during erythropoiesis under levofloxacin treatment --- p.72 / Figure 3.11 Percentage of GPA+ cells in levofloxacin-treated TF-1 erythropoiesis as compared to control --- p.73 / Figure 3.12 Cell death versus mitochondrial transmembrane potential (Δψm) during erythropoiesis under levofloxac in treatment --- p.74 / Figure 3.13 Expression level of antioxidant enzymes during erythropoiesis --- p.75 / Figure 3.14 Expression level of Glut l during erythropoiesis --- p.76 / Figure 3.15 Expression level of Glut l in GPA positive and GPA negative populations --- p.77 / Figure 3.16 Cell death under oxidative stress challenging during erythropoiesis --- p.78 / Figure 3.17 Expression level of antioxidant enzymes and Glutl during erythropoiesis under EUK-134 treatment --- p.79 / Figure 3.18 Expression level of antioxidant enzymes and Glutl during erythropoiesis under vitamin E treatment --- p.80 / Figure 3.19 Cell death under oxidative stress challenging during erythropoiesis under vitamin E treatment --- p.82 / Figure 3.20 Expression level of antioxidant enzymes during erythropoiesis under vitamin C treatment --- p.83 / Figure 3.21 Cell death under oxidative stress challenging during erythropoiesis under vitamin C treatment --- p.84 / Figure 3.22 Cell death under oxidative stress challenging during erythropoiesis under NAC treatment --- p.85 / Figure 3.23 Summary of oxidative stress challenging during erythropoiesis --- p.86 / REFERENCES --- p.87

In this thesis I have connected different evolutionary studies of Lioalemus lizards. In Chapter 1, I followed an integrative approach to delimit species in the Liolaemus walkeri complex. Using mitochondrial markers, morphological data, bioclimatic information and methods appropriate for each data type, we found that the name L. walkeri was covering three new lineages. Three new species were described and one of them (L. chavin) is now categorized as Near Threatened in the IUCN red list. In Chapter 2, I change the subject from species boundaries to the study of viviparity and placentation. In this paper we employed scanning electron and confocal microscopy to compare the placental ultra-structure and pattern of blood vessels in two Liolaemus species. One of the most remarkable traits found is the complete reduction of the eggshell in both placentae, a possible adaptation to improve gas exchange in the hypoxic environments of the high Andes. In chapter 3, I returned to the issue of species delimitation and employed two integrative approaches: a hypothetical deductive framework and a model-based procedure. I applied both approaches in lowland and highland Liolaemus species of the montanus group. I found that in only one case (of four) an unnamed lowland lineage ("Nazca") was delimited concordantly by both procedures. In Chapter 4, I focus on a study of convergent evolution of desert phenotype in Liolaemus species and Ctenoblepharys adspersa. I performed a Bayesian time calibrated and maximum likelihood tree based on 55 taxa and seven molecular markers. We employed quantitative and categorical traits based on 400 specimens and non-metric multidimensional scaling to obtain new quantitative variables. I used three phylogenetic comparative methods to identify and measure the strength of convergence. My results found a strong case of convergent traits in C. adspersa, L. lentus, L. manueli, L. poconchilensis and L. stolzmanni that are probably related to predator avoidance in the Peruvian-Atacama and Monte deserts. In addition, my time calibrated tree resolves the origin of these traits first in C. adspersa at about 80 million years (My) and later independently in Liolaemus species at about 25 My suggesting the present of evolutionary constraints.

Les virus foamy simiens (VFS) sont la troisième famille des rétrovirus complexes exogènes infectant l'Homme. Ces virus, zoonotique, sont transmis par des fluides biologiques (principalement la salive), lors d'un contact direct entre un individu et un singe infecté. Ils établissent une infection chronique chez l'hôte infecté. À ce jour, aucune pathologie n'a été associée à au cas d'infections humaines par le VFS, et aucun cas de transmission secondaire n'a été rapporté dans la population. L'infection VFS représente un modèle naturel de la restriction de l'émergence d'un rétrovirus simien chez l'Homme. Lors de ma thèse, j'ai caractérisé la réponse humorale contre les VFS chez des personnes vivant au Cameroun ou au Gal infectées suite à des morsures lors d'épisodes de chasse. J'ai montré la présence d'anticorps neutralisants les VFS dans 48 sujets infectés, à des titres élevés. La population étudiée est infectée par des virus de deux génotypes qui diffèrent dans le domaine central de la protéine impliqué dans la liaison au récepteur cellulaire. J'ai montré des réponses neutralisantes spécifiques de chaque génotype des sujets étudiés et des réactivités croisées chez 40% d'entre-eux. Parmi ces derniers, la moitié est infectée par deux souches virales. En conclusion, mon travail de thèse est la première caractérisation des anticorps neutralisants chez des personnes infectées chroniquement par un VFS zoonotique : ces anticorps sont fréquemment détectés, à des titres élevés et sont dirigés contre des épitopes conservés entre les VFS de chimpanzé et de gorille. / Simian foamy virus (SFV) are the third family of exogenous complex retroviruses infecting humans. These viruses, of origins, are transmitted by body fluids (mainly saliva), through a direct contact between an individual and an infected m establish a chronic infection in the infected human host. To date, neither pathology, nor secondary transmission has be to be associated with SFV infection in humans. Therefore, SFV represents a natural model of restriction emerging simiar in humans. During my PhD, I characterized the humoral response against SFV in people living in Cameroon and Gabon, mainly infected bites during hunting episodes. I showed the presence of SFV neutralizing antibodies in the plasma of 48 infected individ titers. Our study population is infected with viruses of 2 different genotypes, which differ in the central region of the En region involved in binding to the cellular receptor.While in 60% of cases, neutralizing response was specific to a single genotype, 40% of cases showed cross-reactivity. Cr( was associated in 50% of cases with co-infection with viruses from both genotypes.In conclusion, my PhD is the first study to characterize neutralizing antibodies in individuals chronically infected with a zoonotic SFV : these antibodies are frequently detected at high titers and are directed against epitopes commonly found in chimpanzee and gorilla SFV.

Theoretically, sympatric species must partition resources or space to allow for coexistence. Determining empirically the specific resources each species exploits and species’ interactions (e.g., intra- and interspecific competition) can sometimes be challenging, thus the data are relatively sparse for certain taxa. This paucity of data exists for octopuses. Therefore, I chose to study niches of two sympatric octopuses (Octopus vulgaris and Macrotritopus defilippi) in an intracoastal habitat. Specifically, I assessed (1) spatial distribution of octopus home or “den” space, (2) habitat association, (3) octopus abundance, (4) foraging activity periods, (5) diet, and (6) associated substrates and behaviors used during foraging events. Octopus den locations were marked by GPS to quantify spatial patterns of both species and their spatial relationship to each other. Habitat associations were measured by quantifying photoquadrats of den and surrounding habitats. For foraging activity periods, a video camera was placed near an octopus den for 24-h observation to determine when each octopus species leaves/returns from foraging. Underwater video recording was used to determine associated foraging substrates and behaviors for both species. Prey remains from octopus’ dens and video recordings indicating prey consumption were used to determine diets of the two octopus species. Video recordings from the 24-h camera and foraging behavior events also provided observations of intra- and interspecific interactions. Results revealed that the two species are interspersed throughout the shallow Florida lagoon and are both abundant during the spring months (March, April, May). Although both species are interspersed throughout the lagoon, their den and surrounding habitat association differed. O. vulgaris was associated with hard bottom and M. defilippi was associated with soft bottom, thus they may not compete strongly for habitats. Each species used different foraging strategies and different primary prey, which may also lessen competition and facilitate coexistence. O. vulgaris had peak foraging activity during night hours, foraged mostly on hard bottom and mainly consumed bivalves while M. defilippi had peak foraging activity during day hours, foraged mostly on soft bottom and mainly consumed crustaceans. Octopuses also had species-specific foraging behaviors, with O. vulgaris using parachute attack and M. defilippi using flounder swimming and tripod stance. Additional intra- and interspecific interactions were video recorded and included: fishes following octopuses, predation attempts, agonistic encounters, cannibalism, and tactile communication. This study identified ecological and behavioral components that may facilitate coexistence of these sympatric species, provided insight into cephalopod niches and ecology, and provided baseline conservation requirements for sand-dwelling cephalopods, both of which may be using this site as a mating and nursery habitat. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2019. / FAU Electronic Theses and Dissertations Collection

Because of the scarcity of information for bird communities at the major mainland rivers of southeastern Alaska, the main objective of this study was to provide baseline information including distribution, status, and habitat associations of breeding birds. I conducted a meta-analysis of all known reports (including the current study) conducted at major mainland rivers during the breeding season. I described bird species composition, distribution, abundance estimates, status, habitat associations, and guild membership for all birds recorded at 11 major mainland rivers. Based on incidental observations, 170 species were recorded by all studies. Of these, 134 species were known or suspected to breed, accounting for 50% of all birds known from Alaska and 80% of all birds known from southeastern Alaska. In addition, I provided information on species of management concern as well as management implications and recommendations. I used point counts to survey birds within deciduous riparian vegetation at 6 major mainland rivers during 2000-2002. I compared bird species composition, abundance, richness, and diversity among four main vegetation types of deciduous riparian vegetation: shrubland, young deciduous forest, mature deciduous forest, and mixed deciduous-coniferous forest. Species richness was similar among all habitat types; however, relative abundance and diversity of birds was highest in mixed forest stands. Mature forests had the greatest number of species associated with the Canadian interior. I also used point counts to compare bird species composition, abundance, richness, and diversity among 6 major mainland rivers consisting of three trans-mountain and three coastal rivers. Latitude, connectivity, and availability of mature and mixed forests were the major factors thought to cause differences in bird communities among rivers. Contrary to our predictions, coastal rivers had higher bird species richness, diversity, point abundance , and point richness than trans-mountain rivers. Of the 10 species associated with the Canadian interior recorded during point counts, 8 occurred at both trans-mountain and coastal rivers.

For over 40 000 years, the Australian Aboriginal people relied on native plants as a source of medicinal agents. Although they employed a wide range of species for this purpose, some of the most commonly used were Eremophila species (Myoporaceae). In particular, E. alternifolia was considered the 'number one medicine'. Reportedly, infusions of the leaves were used both internally and externally as an analgesic, decongestant and expectorant. This was believed to be useful in the treatment of colds, influenza, fevers and headaches, as well as for sterilising septic wounds and for promoting general well being.A methanolic extract of E. alternifolia leaves was tested on Langendorff rat hearts and was shown to mediate significant increases in heart rate (chronotropism), contractile force (inotropism) and coronary perfusion rate (CPR). With similar responses to those induced by adrenaline, it seemed likely that the active constituents of E. alternifolia leaves may have acted through adrenergic receptors. Challenging the extract with alpha- and beta- receptor antagonists did not, however, significantly reduce the size of responses. The active compound acted through a different mechanism.The identity of the active constituent in E. alternifolia leaves was revealed and shown to be a known phenylethanoid glycoside called verbascoside. Also known as acteoside, this compound's identity was confirmed by comparison of its spectral parameters with those described in the literature and by comparative tlc behaviour with that of standard samples. Its effects on the Langendorff rat heart were similar to those induced by the methanolic extract, but were more pronounced and did not act through adrenergic receptors or slow calcium channels . Repeatedly, 1 ml of 1 mM verbascoside dose-dependently increased chronotropism, inotropism and CPR in Langendorff rat hearts by increasing ++ / intracellular levels of the second messenger, cyclic, 3',5'-adenosine monophosphate (cAMP). The increase in cAMP production occurred in response to an increase in prostacyclin. Significant increases in this hormone-like compound were detected in hearts treated w verbascoside. The calcium channel blocker, verapamil, did not significantly diminish the effects of verbascoside.Another of the highly prized medicinal agents employed by the Aboriginal people was E. longifolia, a plant of sacred and mystical significance. Infusions made from the leaves were prepared for eye washes, as counter-irritants, for treating headaches and for skin and body washes. Pharmacological testing of the methanolic extract of leaves indicated that one or more active compounds with a biphasic effect on the Langendorff rat heart were responsible. The major active constituent was identified as an iridoid glucoside called geniposidic acid. One ml of 1 mM geniposidic acid significantly inhibit chronotropism, inotropism and CPR. The compound did not, however, induce biphasic effect. This was later rationalised by the small amounts of verbascoside that co-occur in the extract.Four similar iridoid glucosides that had been isolated from three other Eremophila species were tested for their effects on Langendorff rat hearts. Melampyroside (from E. pantonii), verminoside (E. ionantha), ferruloylajugol (E. pantonii) and catalpol (E. maculata subsp brevifolia) all significantly altered myocardial activity in the isolated rat heart preparations. Melampyroside and verminoside were predominantly stimulatory, while ferruloylajugol was biphasic and catalpol only inhibitory. The effects of these iridoids is discussed in comparison with geniposidic acid and other known cardioactive iridoid glucosides. The results suggest that the effects exhibited by these compounds is more common phenomenon than was previously ++ / realised.

This thesis is prepared in three parts; the first part is a study of the ant species of the southern Carnarvon Basin, which was undertaken in order to determine the patterns of ant species distribution in this arid zone area. The distribution patterns were looked at in terms of biogeographical regions and they demonstrated the transitional nature of this particular area. Recommendations to alter the border between the South-west Province and the Eremaean Province were supported. The next chapter of this thesis analysed ant species from long unburnt and burnt areas of three main vegetation types (two Triodia species grasslands and Acacia aneura woodlands) in the Gibson Desert Nature Reserve. This study was carried out to observe the recovery of ant populations after fire. The results provided further evidence that invertebrates are measurably impacted by fire in the arid zone. The final chapter is a comparison of these two arid zone studies with six other ant community studies from throughout Western Australia. It demonstrated the uniqueness of some arid zone sites as well as related each study to each other according to their ant communities.

An examination was undertaken of the literature and restoration cases for 4 major Australian vegetation types (sclerophyll; rainforest; grassland; and wetland) to explore the proposition that ecological resilience may govern recovery after anthropogenic damage, and/or provide a fundamental guide and measure of success for ecological restoration. Also, primary data were collected from highly degraded sites (5 sclerophyll, 3 rainforest, and 4 grassy sites) to assess recovery after restoration treatment. These were supplemented with questionnaire data from practitioners working at a wider range of rainforest and sclerophyll sites, and reports from practitioners working on grassland and wetland sites. In all 4 vegetation types, species generally fell into two main groups : longer-lived 'resprouters' and shorter-lived 'obligate seeders'. But different resilience models were identified for the 4 vegetation types. The sclerophyll type exhibited higher in situ resilience but lower migratory resilience than the rainforest type, which was facilitated by flying frugivore dispersal to perch trees. Self-perpetuation was more tightly coupled with disturbance in the sclerophyll, grassland and wetland types than rainforest; and therefore 'designed disturbance' played a more obvious role in enhancing recovery within these types, than in rainforest. Results suggest that resilience (as both an ecosystem property and a theoretical concept) is fundamental to the practice of ecological restoration. Some prediction of resilience potential of particular degraded sites (and prediction of the degree and type of restoration subsidy needed) can be based on knowledge of : individual species' recovery mechanisms; resilience models for individual vegetation-types; and the site's colonisation potential and impact history / Doctor of Philosophy (PhD)

The problem of invasive alien species (IAS) is recognized as the second most serious threat to loss of biodiversity after habitat destruction. It is a problem largely created by humans as they transport and introduce species, deliberately and accidentally, from one part of the globe to another. The pressures exerted on biodiversity by international trade are one of the most serious aspects of the IAS problem. Although states are under obligations in international environmental law to prevent the entry of, and control, those alien species that threaten biodiversity, to date state practice has often been found wanting. In particular, quarantine regulation, which can be a state??s first line of defence against IAS, is mainly used by states to protect their farming and agricultural product sectors rather than biodiversity at large. The reasons for this include lack of domestic resources and lack of guidance at the international level. However, even if states were to expand the purview of quarantine, the question arises whether they would be able to use quarantine regulation to protect biodiversity from IAS while simultaneously fulfilling their international trade law obligations. This study seeks to answer this question by examining international environmental law and international trade law in their application to quarantine regulation. In doing so, the study identifies many areas of conflict. The different policies that underpin environmental and trade regimes mean that environmental concepts, such as the precautionary principle and the ecosystem approach, are difficult to apply within the international trade law regime. A way of achieving a more harmonized international response to the problem of IAS is suggested by incorporating environmental considerations into the international standards used by states to design and implement domestic quarantine measures. To facilitate the practical implementation of international standards the study further recommends appropriate financial and institutional capacity building mechanisms.