In ihrer stammesgeschichtlichen Entwicklung haben, neben Säugetieren, Vögeln und Insekten, auch Froschlurche ein umfangreiches Repertoire an akustischen Signalen zur Identifizierung arteigener Individuen und anlocken potenzieller Paarungspartner entwickelt. Diese sogenannten Anzeigerufe sind dabei für jede Art einzigartig. Beobachtungen zeigen aber, dass es neben dieser Artspezifität, auch Ähnlichkeiten in den Anzeigerufen zwischen nicht näher verwandten Arten existieren. Es stellt sich daher die Frage woher diese Ähnlichkeiten stammen. Stellen diese Ähnlichkeiten eine Anpassung an die Bedingungen der Lebensräume dar (konvergente Evolution) oder sind sie ein Ergebnis der stammesgeschichtlichen Entwicklung der Arten (divergente Evolution). Zur Klärung dieser Frage habe ich Anzeigerufe von ca. 1500 Arten aus den unterschiedlichsten Lebensräumen unseres Planten gesammelt und ausgewertet. Eine Korrelation der gemessenen akustischen Eigenschaften (z. B. dominante Frequenz) mit der Phylogenie der Amphibien sollte dabei klären, inwieweit die Phylogenie mit diesen akustischen Eigenschaften verknüpft ist. Vergleiche zwischen verschiedenen Kategorien aus Morphologie, Verhalten und Lebensraum in Bezug auf die akustischen Eigenschaften wiederum sollten mögliche Anpassungen der akustischen Eigenschaften herausstellen. Es stellte sich am Ende heraus, dass einige akustische Eigenschaften (z. B. dominante Frequenz) sowohl ein Ergebnis einer divergenten Evolution sind, als auch einer konvergenten Evolution. Zusätzlich war es mir möglich die Anzeigerufe in separate Gilden einzuteilen, die ähnliche strukturellen Eigenschaften aufweisen. Gilden in die Arten unabhängig von ihren verwandtschaftlichen Beziehungen eingeordnet werden können. Ein Werkzeug was Vergleiche zwischen Arten anhand ihrer Akustik vereinfacht und die Möglichkeit gibt zu verstehen wie Arten die Bedingungen ihrer Lebensräume auf ähnliche akustische Weise nutzen. / Beside mammals, birds and insects also anurans developed at their evolutionary history a wide repertoire of acoustic signals to identify conspecific individuals and to attract possible mating partners. These so-called advertisement calls are unique to every single species. However, beside this species-specific uniqueness we can also observe astonishing similarities among advertisement calls of not close related species. Therefore, there is the question of where these similarities came from. Are these similarities adaptations to conditions to similar biology’s and environments and so a result of a convergent evolution or these similarities arose by the anuran phylogeny and represent a divergent evolution? To answer these questions I collected and measured advertisement calls of ca. 1500 anuran species around the globe of different living habitats. A correlation between phylogeny and acoustic properties (e.g. dominant frequency) could show the influence of the phylogeny at the evolved acoustic traits. Comparisons among different categories of morphology, behaviour and habitat concerning the acoustic traits could show possible adaptations. The result of my research showed that some structural and spectral properties are a consequence of the phylogenetic history as well as an adaptation to conditions of biology and environment. Both, the divergent and the convergent evolution place a role in these cases. Additional I managed to introduce a system to order anuran advertisement calls into distinct guilds. These guilds show how different species uses their acoustic properties in a similar matter. An additional tool to make comparisons among species easier and to understand how anuran uses environments in a similar acoustic way.

Im Bericht wird bisher ungeklärten Fragen des Übertragungswegs des Koi-Herpesvirus nachgegangen. In latent infizierten Laichkarpfenbeständen, in deren Geschlechtsprodukten und in befruchteten Eiern lässt sich das Virus (KHV) nachweisen. Eine Infektion der frisch geschlüpften Karpfenbrut scheint allerdings nicht zu erfolgen. Brütlinge von Laichfischen, die aus KHV-positiven Beständen stammen, haben eine höhere Resistenz gegenüber einer erneuten Infektion mit dem KHV. Der Beitrag richtet sich an praktische Fischzüchter, Fachtierärzte, Fischereiwissenschaftler und an die interessierte Öffentlichkeit. Redaktionsschluss: 31.03.2022

The plants and animals that inhabit river channels may act as zoogeomorphic agents affecting the nature and rates of sediment recruitment, transport and deposition. The impact of benthic-feeding fish, which disturb bed material sediments during their search for food, has received little attention, even though benthic feeding species are widespread in rivers and may collectively expend significant amounts of energy foraging across the bed. A series of experiments were conducted to investigate the impacts of benthic feeding fish on the structure and composition of gravel-bed river sediments, and the implications for bed material transport. An ex-situ experiment was conducted to investigate the impact of a benthic feeding fish (European Barbel Barbus barbus) on particle displacements, bed sediment structures, gravel entrainment and transport fluxes. In a laboratory flume, changes in bed surface topography were measured and grain displacements examined when an imbricated, water-worked bed of 5.6-16 mm gravels was exposed to feeding juvenile Barbel. For substrates that had been exposed to feeding fish and control substrates which had not, grain entrainment rates and bedload fluxes were measured under a moderate transport regime. On average, approximately 37% of the substrate, by area, was modified by foraging fish during a four-hour treatment period, resulting in increased microtopographic roughness and reduced particle imbrication. Structural changes caused by fish increased bed load flux by 60% under entrainment flows, whilst on average the total number of grains transported during the entrainment phase was 82% higher from substrates that had been disturbed by Barbel. An ex-situ experiment utilising Barbel and Chub Leuciscus cephalus extended this initial study by considering the role of fish size and species as controls of sediment disturbance by foraging. Increasing the size of Barbel had a significant effect on measured disturbance and bedload transport. Specifically, the area of disturbed substrate, foraging depth, microtopographic roughness and sediment structure all increased as functions of fish size, as did bedload flux and total transported mass. In a comparison of the foraging effects of like-sized Barbel and Chub 8-10 in length, Barbel foraged a larger area of the riverbed and had a greater impact on microtopographic roughness and sediment structure. Foraging by both species was associated with increased sediment transport, but the bed load flux after foraging by Barbel was 150% higher than that following foraging by Chub and the total transported mass of sediment was 98% greater. An in-situ experiment quantified the effects of foraging fish, primarily Cyprinids (specifically Barbel and Chub), on gravel-river bed sediment structures, surface grain-size distributions, sediment transport fluxes and grain entrainment in the River Idle, Nottinghamshire, UK. This was achieved by installing large experimental sediment trays seeded with food at typical densities. The experiments yielded data about 1) topographic and structural differences between pre- and post-feeding substrates using DEMs interpolated from laser scans, 2) modifications to surface and sub-surface grain-size distributions as a function of fish foraging and 3) differences in sediment entrainment from water-worked substrates exposed to feeding fish and control substrates, without fish. Small sections of the substrate trays were recovered in tact from the field and for substrates that had been exposed to feeding fish and control substrates which had not, grain entrainment rates and bedload fluxes were measured under a moderate transport regime in the laboratory. On average, approximately 74% of the substrate, by area, was modified by foraging fish during a twelve-hour period, resulting in increased microtopographic roughness and substrate coarsening which had significant implications for bed material transport during the steady entrainment flow. Together, results from these experiments indicate that by increasing surface microtopography, modifying the composition of fluvial substrates and undoing the naturally stable structures produced by water working, foraging can influence sediment transport dynamics, predominately by increasing the mobility of river bed materials. The implication of this result is that by influencing the quantity of available, transportable sediment and entrainment thresholds, benthic feeding may affect sediment transport fluxes in gravel-bed rivers. In addition, three discrete studies were performed alongside the core experiments described above. A quantitative examination of habitat conditions favoured by feeding Barbel was conducted in the River Idle (Nottinghamshire, UK) which served to supplement existing literature pertaining to Barbel ecology, and inform experimental design during the core experiments. Two further studies considered the potential importance of foraging as a zoogeomorphic activity in terms of spatial extent, at a variety of scales, thereby extending core experiments to larger spatial scales in-situ.

This thesis presents an investigation into Gyrodactylus species recognition, making use of machine learning classification and feature selection techniques, and explores image feature extraction to demonstrate proof of concept for an envisaged rapid, consistent and secure initial identification of pathogens by field workers and non-expert users. The design of the proposed cognitively inspired framework is able to provide confident discrimination recognition from its non-pathogenic congeners, which is sought in order to assist diagnostics during periods of a suspected outbreak. Accurate identification of pathogens is a key to their control in an aquaculture context and the monogenean worm genus Gyrodactylus provides an ideal test-bed for the selected techniques. In the proposed algorithm, the concept of classification using a single model is extended to include more than one model. In classifying multiple species of Gyrodactylus, experiments using 557 specimens of nine different species, two classifiers and three feature sets were performed. To combine these models, an ensemble based majority voting approach has been adopted. Experimental results with a database of Gyrodactylus species show the superior performance of the ensemble system. Comparison with single classification approaches indicates that the proposed framework produces a marked improvement in classification performance. The second contribution of this thesis is the exploration of image processing techniques. Active Shape Model (ASM) and Complex Network methods are applied to images of the attachment hooks of several species of Gyrodactylus to classify each species according to their true species type. ASM is used to provide landmark points to segment the contour of the image, while the Complex Network model is used to extract the information from the contour of an image. The current system aims to confidently classify species, which is notifiable pathogen of Atlantic salmon, to their true class with high degree of accuracy. Finally, some concluding remarks are made along with proposal for future work.

Zebrafish (Danio rerio) have been extensively used over the past two decades to study muscle development, human myopathies and dystrophies, due to its higher degree of homology with human disease causing genes and genome. Despite its unique qualities, zebrafish have only been used as an in-vivo model for muscle development research, due to the limitations surrounding lack of a consistent isolation and culture protocol for zebrafish muscle progenitor cells in-vitro. Using different mammalian myoblast isolation protocols, a novel and robust protocol has been developed to successfully isolate and culture zebrafish skeletal muscle cells repeatedly and obtain differentiated long multi nucleated zebrafish myotubes. Commitment to myogenic lineage was confirmed by immuno-staining against muscle specific protein desmin, and expression pattern of different genetic markers regulating myogenesis. In order to recapitulate the in-vivo bio-physiological environment for zebrafish skeletal muscle cells in-vitro, these cells were successfully cultured in tissue engineered three dimensional (3D) constructs based on fibrin and collagen models. Maturation of tissue engineered collagen and fibrin based constructs was confirmed using the basic parameters described in the literature i.e. collagen three times greater contraction from the original width (Mudera, Smith et al. 2010) and fibrin constructs tightly coiled up to 4mm of diameter (Khodabukus, Paxton et al. 2007). In-vitro characterisation of zebrafish skeletal muscle cells showed hypertrophic growth of muscle mass compared to hyperplasic growth in-vivo as suggested for fish species in literature (Johnston 2006), which is different from human and other mammals. Comparative analysis of zebrafish muscle cells cultured in monolayer against cultured in 3D tissue engineered constructs showed significant increase in fusion index, nuclei per myotube (two-fold) and myotubes per microscopic frame (two-fold). Cells cultured in tissue engineered construct closely resembled in-vivo muscle in terms of their unidirectional orientation of myotubes. These tissue engineered 3D zebrafish skeletal muscle models could be used for various purposes such as drug screening, effect of different temperature extremes, studying underlined pathways involved in human diseases; and with further refinements it would potentially replace the need for studies on live fish in these areas.

The mangrove killifish, Kryptolebias marmoratus, has the potential of becoming a strong model organism for a range of biological disciplines thanks to its ability to self-fertilise, a process only known to occur in invertebrate animals until its discovery. Selfing, a natural occurrence in this species, has lead to the formation of clonal lineages composed of highly homozygous individuals. The aim of this thesis was to further establish K. marmoratus in the field of developmental biology by providing an information infrastructure to help advance research on this peculiar animal and further promote its place in the pantheon of model organisms. To do so, I first set out to standardise K. marmoratus embryology by providing defined developmental stages with clear visual representations of key embryonic structures. This staging series is an essential tool that will ensure repeatability and consistency within and across different laboratories. Secondly, I examined several techniques for embryonic manipulation and for imaging that can be used in an array of experimental designs. Using these techniques I demonstrated microinjection of embryos by monitoring the yolk syncytial layer and its nuclei, and time-lapse analyses of the yolk surface during embryonic development. Finally, I applied the knowledge gained from my first two studies and examined Bmp signalling in K. marmoratus embryos and its influence on body patterning. By inhibiting this pathway, I found a new phenotype characterised by an extremely short and split body axis. These data highlighted the importance of studying known signalling pathways in unknown organisms as species-specific differences may improve our understanding of fundamental developmental processes. This thesis demonstrates that with its easily obtainable and manipulated embryos, K. marmoratus can be used for embryological research in the same light as other model organisms such as zebrafish or medaka. The rising amount of information on mangrove killifish will help further take advantage of this unique and intriguing species, and supports the use of this hermaphroditic vertebrate as a strong comparative model in developmental biology.

Correct species identification is fundamental to all areas of biology, but particularly the policy related areas of conservation and fisheries management. To enable guidelines to be developed for environmental management and conservation, such identifications need links to studies of the evolutionary history, biological factors and environmental influences driving species divergence and population dynamics for the target species. This study concerns two genera of gadiform fish, Muraenolepis and Macrourus, found in southern temperate and Antarctic waters, with a single species, Macrourus berglax, present in the North Atlantic. With similar distribution patterns to toothfish species, Dissostichus eleginoides and D. mawsoni, they are a major food source and by-catch of the toothfish fishery. Both are slow growing and long lived, with different evolutionary histories, life expectancies and strategies for reproduction. For both genera, the accuracy of morphological keys, number of described species and their distribution is under debate. This study has identified specimens to species level using both morphological and genetic techniques, redefining the range for morphological features and taxonomic keys. For Muraenolepis, this has clarified confusion over Mu. marmoratus and Mu. microps being a single species, confirmed some mis-identification from sexual dimorphism and provided genetic evidence for the recently described species Mu. evseenkoi. For Macrourus, this work has identified a new species, now named Ma. caml, and found that Ma. holotrachys and Ma. berglax are genetically identical, raising the question of bipolar distribution or recent divergence. The low level of genetic variation within both species suggests a recent evolution and expansion into Antarctic waters. Similar geographic species limits imply common processes influencing divergence, with the oceanographic fronts as potential barriers. Further investigation of niche overlap and fine scale population structure are required to fully understand the processes driving speciation and provide the underlying data required for fisheries management.

La diadromie représente un cycle de vie qui implique obligatoirement des migrations entre la mer et l’eau douce. La diadromie s’est maintenue dans différentes lignées évolutives de poissons, mettant ainsi en évidence l’existence d’avantages adaptatifs comme la capacité à coloniser des milieux vierges ou de mieux exploiter la variabilité spatio-temporelle de l’environnement. Une hypothèse pour expliquer l’origine et le maintien de la diadromie postule que les migrations entre mer et rivière ont pour but de placer les individus dans le milieu qui favorisera la croissance avant la première reproduction. Une croissance en mer et une reproduction en rivière (anadromie) serait favorisée si la productivité primaire en eau douce est inférieure à celle des milieux marins environnants tandis que la situation inverse (catadromie ; reproduction en mer, croissance en rivière) est attendue quand le différentiel de productivité est en faveur des eaux douces. Les résultats de notre étude réalisée à l’échelle du globe confirment cette hypothèse et montrent également que les poissons diadromes se trouvent majoritairement dans des rivières à faible richesse spécifique et/ou ouvertes à la colonisation après le retrait des glaciers au Quaternaire. Leur capacité à coloniser de nouveaux milieux permet aux diadromes d’être très présents sur les îles océaniques. Si la biodiversité sur les îles océaniques a été largement étudiée, très peu d’études ont porté sur les poissons d’eau douce qui s’y trouvent. Notre étude des peuplements de poissons des îles polynésiennes confirme la généralité d’un modèle proposé pour les faunes terrestres prédisant une relation en dôme entre richesse spécifique et âge de l’île. Nos résultats montrent également que la richesse diminue avec la distance au centre de biodiversité le plus proche et augmente avec l’altitude maximale de l’île.Malgré leur succès évolutif, les poissons diadromes se révèlent très vulnérables face aux activités humaines (surexploitation, pollution de l’eau, barrages…). De ce fait, de nombreuses espèces sont classées comme vulnérables par l’IUCN. Paradoxalement, notre étude sur les caractéristiques biologiques et écologiques des espèces qui se sont éteintes récemment à l’échelle du globe n’identifie pas la diadromie comme un facteur aggravant. Cette étude montre que le critère biologique principal menant à l’extinction est l’endémisme et une aire de distribution géographique restreinte. Bien que souvent en déclin, beaucoup d’espèces diadromes ont jusqu’ici échappé à l’extinction du fait de leur aire de distribution initiale de grande taille, résultat probable de leur grande capacité de dispersion. Un très bon exemple est l’esturgeon européen, Acipenser sturio, initialement largement distribué en Europe mais qui après un fort déclin à partir de 1850 ne se reproduit plus que dans le seul bassin de la Garonne. Pour mieux suivre le déclin de cette espèce au cours du temps, nous avons étudié sa distribution avant 1850 à l’aide de données archéozoologiques. L’analyse de ces données suggère une diminution des populations très ancienne, initiée il y a 2500 ans. A l’aide de températures passées reconstituées, nous avons montré que des températures élevées expliquent en partie l’occurrence d’esturgeons dans les sites archéologiques mais sans contribuer à expliquer son déclin. Ces résultats suggèrent un impact de l’homme précoce sur les populations d’esturgeons. Cette thèse a permis de synthétiser les connaissances sur les poissons diadromes concernant leur distribution géographique à diverses échelles de temps et d’espace, et de contribuer à une meilleure compréhension de leur diversité et de leur vulnérabilité. / Diadromous species exhibit a life-cycle implying migrations between freshwater and ocean. Diadromy is observed in many fish lineages suggesting that adaptive traits are associated with this strategy such as better dispersal ability and more efficient tracking of the spatio-temporal variability of the environment. One of the hypotheses that have been put forward to explain the evolution and persistence of diadromy states that the purpose of migrations is to select environmental conditions that will optimize pre-reproductive growth. Growing at sea and reproducing in river (anadromy) is supposed to be favored when freshwater primary productivity is lower in freshwater than in the nearby sea. The reverse (growing in freshwater, reproducing in ocean; catadromy) is expected when productivity is higher in sea than in freshwater. According to our study, conducted at a global scale, this hypothesis holds true. In addition it is shown that diadromous species are overrepresented in species poor rivers and/or in those that have been open to colonization after the retreat of glaciers after the last glacial maximum. Because of their dispersal ability, diadromous species are also over-represented on oceanic islands. Biodiversity on islands is classical topic of biogeography but yet few studies have dealt with freshwater fishes. In our study of fish communities from the Polynesian islands, we demonstrate the generality of a model initially built to explain terrestrial biodiversity on oceanic islands. As predicted by this model, species richness peaks at peaks at intermediate island age. Also revealed by our results are a positive relationship between island elevation and species richness and a decrease in species richness as the distance from the nearest biodiversity hotspot increases. Diadromy has been a successful strategy over geologic times but in face to human activities it seems to contribute to extinction proneness, as exemplified by the numerous diadromous species red listed by IUCN. Paradoxically, our study about the biological and ecological traits shared by the freshwater fishes that have been recently globally extinct does not suggest that diadromy has been a major factor. This study points to endemism and restricted geographic range size as the major determinants of extinction. It seems that because of their dispersal abilities, diadromous species tend to have widespread historical distributions and, even if many species are declining, this prevented most of them from being entirely extirpated. This is well exemplified by the European sturgeon (Asipenser sturio), formerly widely distributed over Europe and now restricted to one reproducing population in the Garonne River, because of dramatic population collapses after 1850. To have a better idea of the fate of this species before 1850, we analyzed archaeozoological records. According to our analyses, sturgeons started to decline a long time ago, about 2500 years ago. Using reconstructed past temperatures, our analyses show that a high temperature increases the probability of finding this species in archeological remains but cannot explain entirely the observed decline. This suggests that human activities impacted sturgeon populations well before the industrial revolution. This thesis by synthesizing species occurrence data over different spatial and temporal scales contributed to a better knowledge about the diversity and vulnerability of diadromous fishes.

Bibliography: leaves 129-158. / xvii, 159, [11] leaves, [24] leaves of plates : ill. (some col.) ; 35 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / The major estrogen inducible protein in zebra fish liver has been purified to homogeneity by FPLC using anion exchange chromatography (Mono-Q Pharmacia) with purification being monitored by SDS-PAGE electrophoresis. / Thesis (Ph.D.)--University of Adelaide, Dept. of Obstetrics and Gynaecology, 1995?

Despite teleosts being the largest and most diverse group of vertebrates, fish models currently used to study photoperiodic effects on fish physiology have been limited to a few species, most of which are temperate seasonal breeders. The overall aim of this work was to expand our knowledge on circadian biology and environmental physiological effects in Nile tilapia (Oreochromis niloticus niloticus), a continuous breeding species of tropical-subtropical origin. The circadian light axis of Nile tilapia is described with regards to melatonin production. Circadian melatonin profiles of fish under 12L:12D photoperiods were observed to be low at day and high at night, suggesting melatonin to be an entraining signal as observed in all other vertebrates. When constant light (LL) was used, such day and night fluctuations where abolished. However when fish where exposed to constant darkness (DD) a strong robust endogenous melatonin rhythm was found, suggesting the presence of circadian oscillators in this species. Importantly, this endogenous rhythm was observed to be maintained for at least three weeks under darkness and proved to be circadian in nature. Moreover, although the melatonin system was able to produce day and night melatonin rhythms when exposed to a different (6L:6D) photocycle, the oscillator appeared to not be entrainable to such a short photo cycle when exposed to DD, as melatonin levels remained high. When comparing the circadian organization of different teleost species including Nile tilapia, preliminary studies showed at least three divergent circadian light organizations in teleosts. Nile tilapia was characterised by a pineal gland far less sensitive than in other fish species as demonstrated through in vitro studies. Furthermore, pineal melatonin production was clearly dependent on the light perceived by the eyes as ophthalmectomy resulted in basal plasma melatonin levels during the dark period. These findings are the first to be reported in a teleost and could be comparable to the circadian light organization of higher vertebrates such as mammals. The onset of puberty of Nile tilapia was studied with regards to the newly discovered Kiss1/GPR54 system. Such a system has recently been discovered in mammals and found to be the primary switch of the brain-pituitary-gonadal (BPG) axis. The results of this study not only suggest a link between the Kiss1/GPR54 system and the onset of III puberty in this tropical batch spawning teleost, that would be a highly conserved feature across vertebrates, but also that the transcriptional mechanisms regulating GPR54 expression could be directly or indirectly influenced by light. Finally, a study was conducted on the effects of different intensities of continuous light (LL) on the growth and sexual development of Nile tilapia up to first maturation. The results showed a significant growth response of fish in all LL treatments compared to control fish. Importantly, this confirmed that LL enhances growth in this species and suggests that it is the light regime more than the intensity which is having an effect. This work thus provides important basic knowledge of the light entrainment pathway and circadian melatonin rhythms in Nile tilapia. Of special importance is the discovery of a strong endogenous melatonin oscillator and a novel circadian organization in fish which would seem to be homologous to that observed in higher vertebrates. Moreover, this work provides evidence that the newly discovered Kiss1/GPR54 system has a similar role in fish as has been found in mammals and that such a system could be directly or indirectly regulated by light. If so, Nile tilapia and other fish species could become important models in the chronobiology and reproduction fields. Finally, this work not only increases our basic and applied knowledge of this species, but also broadens our understanding of the circadian light axis in teleosts and its mediatory effects on reproduction.