付記する学位プログラム名: 霊長類学・ワイルドライフサイエンス・リーディング大学院 / 京都大学 / 新制・課程博士 / 博士(理学) / 甲第25154号 / 理博第5061号 / 新制||理||1721(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 平田 聡, 教授 松田 一希, 准教授 半谷 吾郎 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

Individuals within fish populations differ in many traits, such as sex, life-history, habitat residence, diet, and morphology. Such trait differences among individuals (i.e. intra-population variation) may be greater than the differences among populations (i.e. inter-population variation). My dissertation examines intra-population variation, with a focus on trophic relationships and morphology; as well as how variation in these attributes may reflect differences in bioaccumulation of contaminants. The second chapter of my dissertation examines the influence of spatial-temporal variation on the trophic structures of round goby (Neogobius melanstomus) and two age classes of yellow perch (Perca flavescens) within Saginaw Bay, Lake Huron. Using stable isotope ratios (δ13C, δ15N, δ2H, δ18O) and stomach contents as trophic indicators, I examined variation of diets. I found that spatial variation had a greater impact on diet indicators than both annual and seasonal variation. This spatial variation could represent a form of compartmentalization within the community of fish residing in Saginaw Bay, and could provide stability to the community. Chapter three of my dissertation examines intra-population variation in yellow perch morphology through a series of mesocosm experiments. My first mesocosm study determined that yellow perch could be experimentally manipulated to display divergent morphologies using simulated habitats, specifically pelagic and littoral habitats. Following this experiment, I focused on specific environmental drivers (structure, prey resources, and predation risk) as possible influences on yellow perch morphology. Within experimental pools, I exposed yellow perch to one of four treatments (an open pool, a structured pool, pools with chironomid prey resources and pools with a perceived, olfactory, predation risk) in the summer of 2015. Following exposure to these treatments I examined the morphological changes in yellow perch in magnitude and direction. I observed that while each treatment induced some difference in morphology, the open and structured treatments had the greatest magnitude of difference. I repeated the open and structure treatments during the following summer (2016). Again, I found that structure and open morphologies could be induced by my mesocosm treatments, but also observed that shapes differed from the previous year’s structure and open treatments. Finally, my fourth chapter examined how variation in trophic niches and morphology may reflect variation in contaminant concentration of fish in their natural environment. In this chapter, I extended my work with yellow perch to also include black crappie (Pomoxis nigromaculatus) and examined fish from 5 northern Indiana glacial lakes. Using model inference techniques, I found that variation in mercury was closely associated with not only fish total length, but also stable isotopes (δ13C and δ15N) and morphology. Interestingly, morphology-related variables of both species were strong predictors of mercury concentration in fish, following total length. Together, the chapters within my dissertation highlight the importance of considering intra-population variation, in which local factors such as habitat conditions and prey availability can influence individual variation in trophic structuring and morphology. These in turn may reflect other attributes of interest, such as the accumulation of contaminants.

A associação entre distúrbios metabólicos e carência dietética de zinco mostra-se relacionada com o desequilíbrio de nutrientes na dieta, apontado como um fator interferente na biodisponibilidade desse mineral. Este trabalho teve por objetivo, avaliar a biodisponibilidade do zinco em dieta brasileira, adequada em energia e macronutrientes, utilizando técnica com isótopos estáveis. Depois de identificado o perfil alimentar de 12 indivíduos, do sexo masculino, com idade entre 19 e 42 anos, o grupo consumiu, durante 7 dias, uma dieta experimental, equilibrada e variada. No 8° dia do experimento cada indivíduo recebeu uma dose intravenosa de 70Zn e a refeição do almoço marcada, extrinsecamente, com 67Zn, ambos enriquecidos. Os voluntários, com 68 ± 5kg e 1,73 m, apresentavam-se eutróficos (IMC=23 ± 1) e clinicamente saudáveis, tendo-se constatado um consumo usual de dieta hiperproteica e hiperlipídica, para a maioria do grupo. A análise direta das dietas consumidas revelou que foi atendida 95% da recomendação de energia para o grupo (2809 ± 198,54 kcal), distribuída em 16, 26 e 58% para proteínas, lipídios e carboidratos, respectivamente. Ao contrário do Ca (712 ± 79 mg), as concentrações de Zn (11,62 ± 1,55mg) e Fe (10,65 ± 1,47mg) superaram as recomendações, e as frações IP5 e IP6 do fitato não foram detectáveis (<0,01). Das análises das concentrações de zinco no plasma, eritrócitos e urina, conclui-se que o estado nutricional, relativo ao zinco, da maioria dos voluntários mostrou-se satisfatório, uma vez estar de acordo com os padrões de referência. O enriquecimento dos isótopos 67Zn e 70Zn na urina de cada indivíduo foi determinado pela técnica de espectrometria de massa por plasma induzido (ICP-MS), revelando percentual de absorção de 30%, com variações entre 11 e 47%. Os resultados indicam que a biodisponibilidade do zinco na dieta avaliada pode ser classificada como sendo de moderada a alta, segundo critérios da OMS. / This research aimed to evaluate the bioavailability of zine in the Brazilian diet, containing the required energy and macronutrients, by the use of a stable isotope technique. Twelve male eutrophic healthy individuals in age between 19 - 42 years were enrolled in the study. The group consumed an experimental diet for a week, corresponding to 95% of the energy requirement (RDA-1989) and composed of 16% protein, 26% fat and 58% carbohydrate. In day eighth of the experiment, an intravenous injection of 70Zn was given to each individual and the meal consumed at lunch was extrinsically labeled with 67Zn, in that both stable isotopes were enriched. The percentage increase, or enrichment, of 67Zn and 70Zn in the urine of each individual was evaluated by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and showed an average zinc absorption of 30%, variation from 11 to 47%, and a bioavailability from modest to high, according to the WHO criteria.

Blooms of nitrogen fixing cyanobacteria (NFC) occur in many freshwater and marine systems, including the Baltic Sea. By fixing dissolved nitrogen, they circumvent general summer nitrogen limitation, while also generating a supply of novel bioavailable nitrogen for non-diazotrophic primary producers and ultimately supporting secondary production. Elucidating trophic links between primary consumers and NFC is essential for understanding role of these blooms for secondary production. However, until recently, there was no reliable method to quantify individual prey species for zooplankter feeding in situ. The development of PCR-based methods to detect prey-specific DNA in the diet of consumers, including microscopic animals, allows identification and quantification of trophic linkages in the field. Using molecular diet analysis in combination with egg production measurements, biochemical markers of growth and condition; and stable isotope approach, we explored a possibility to determine (1) whether cyanobacteria are grazed and assimilated by mesozooplankters (Papers I and II), (2) which species/groups are particularly efficient consumers of cyanobacteria (Papers II and III), and (3) how feeding on cyanobacteria affects zooplankton growth and development (Paper I and III). Taken together, these laboratory and field observations, provided evidence that NFC contribute to feeding and reproduction of zooplankton during summer and create a favorable growth environment for the copepod nauplii (Paper I). The favorable growth conditions for juvenile copepods observed during NFC blooms were hypothesized to be mediated by picoplankton that take up bioavailable nitrogen exuded from cyanobacterial cells. This hypothesis found support in Paper II that provided quantitative estimates for the direct picocyanobacteria → mesozooplankton pathway, with highest weight-specific consumption observed in nauplii. Further, using field observations on zooplankton and phytoplankton development during a growth season in the northern Baltic proper, we found that NFC nitrogen is assimilated and transferred to zooplankton via both direct grazing and indirectly through grazing on small-sized phyto- and bacterioplankton (Paper III). Finally, these and other findings emphasizing the importance of NFC for Baltic Sea secondary production during growth season were synthesized to show that diazotrophic nitrogen enters food webs already at bloom initiation (Paper III) and is transferred via multiple pathways to pelagic and benthic food webs and, ultimately, to fish (Paper IV). / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Accepted.</p>

Food webs are structured by intricate nodes of species interactions which govern the flow of organic matter in natural systems. Despite being long recognized as a key component in ecology, estimation of food web functioning is still challenging due to the difficulty in accurately measuring species interactions within a food web. Novel tracing methods that estimate species diet uptake and trophic position are therefore needed for assessing food web dynamics. The focus of this thesis is the use of compound specific nitrogen and carbon stable isotopes and molecular techniques for assessing predator-prey interactions and energy flow in natural aquatic ecosystems, with a particular focus on the species links between phytoplankton and zooplankton. The use of δ15N amino acid values to predict organism trophic position are evaluated through a meta-analysis of available literature which included measurements from 359 marine species (article I). Through a controlled feeding study isotope incorporation in aquatic organisms, across both plant-animal and animal-animal species linkages is further assessed (article II). These studies showed that δ15N amino acid values are useful tools for categorizing animal trophic position. Organism feeding ecology influenced nitrogen trophic discrimination (difference in isotope ratio between consumer and diet), with higher discrimination in herbivores compared to omnivores and carnivores (article I). Nitrogen isotope trophic discrimination also varied among feeding treatments in the laboratory study (article II). The combined findings from articles I &amp; II suggest that researchers should consider using group specific nitrogen trophic discrimination values to improve accuracy in species trophic position predictions.  Another key finding in the controlled laboratory study (article II) was consistently low carbon isotope discrimination in essential amino acids across all species linkages, confirming that these compounds are reliable dietary tracers. The δ13C ratios of essential amino acids were applied to study seasonal dynamics in zooplankton resource use in the Baltic Sea (article III). Data from this study indicated that zooplankton assimilate variable resources throughout the growing season. Molecular diet analysis (article IV) showed that marine copepod and cladoceran species ingested both autotrophic and heterotrophic resources. Evidence from both articles III &amp; IV also revealed that zooplankton feed on a relatively broad range of diet items but not opportunistically on all available food sources. Mesozooplankton feeding patterns suggested that energy and nutritional flows were channelled through an omnivorous zooplankton food web including microzooplankton prey items. Overall the results of this thesis highlight that stable isotope ratios in specific compounds and molecular techniques are useful tracing approaches that improve our understanding of food web functioning. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: Manuscript. Paper 4: Manuscript.</p><p> </p>

abstract: An understanding of diet habits is crucial in implementing proper management strategies for wildlife. Diet analysis, however, remains a challenge for ruminant species. Microhistological analysis, the method most often employed in herbivore diet studies, is tedious and time consuming. In addition, it requires considerable training and an extensive reference plant collection. The development of DNA barcoding (species identification using a standardized DNA sequence) and the availability of recent DNA sequencing techniques offer new possibilities in diet analysis for ungulates. Using fecal material collected from controlled feeding trials on pygmy goats, (Capra hicus), novel DNA barcoding technology using the P6-loop of the chloroplast trnL (UAA) intron was compared with the traditional microhistological technique. At its current stage of technological development, this study demonstrated that DNA barcoding did not enhance the ability to detect plant species in herbivore diets. A higher mean species composition was reported with microhistological analysis (79%) as compared to DNA barcoding (50%). Microhistological analysis consistently reported a higher species presence by forage class. For affect positive species identification, microhistology estimated an average of 89% correct detection in control diets, while DNA barcoding estimated 50% correct detection of species. It was hypothesized that a number of factors, including variation in chloroplast content in feed species and the effect of rumen bacteria on degradation of DNA, influenced the ability to detect plant species in herbivore diets and concluded that while DNA barcoding opens up new possibilities in the study of plant-herbivore interactions, further studies are needed to standardize techniques and for DNA bar-coding in this context. / Dissertation/Thesis / M.S. Applied Biological Sciences 2012

A associação entre distúrbios metabólicos e carência dietética de zinco mostra-se relacionada com o desequilíbrio de nutrientes na dieta, apontado como um fator interferente na biodisponibilidade desse mineral. Este trabalho teve por objetivo, avaliar a biodisponibilidade do zinco em dieta brasileira, adequada em energia e macronutrientes, utilizando técnica com isótopos estáveis. Depois de identificado o perfil alimentar de 12 indivíduos, do sexo masculino, com idade entre 19 e 42 anos, o grupo consumiu, durante 7 dias, uma dieta experimental, equilibrada e variada. No 8° dia do experimento cada indivíduo recebeu uma dose intravenosa de 70Zn e a refeição do almoço marcada, extrinsecamente, com 67Zn, ambos enriquecidos. Os voluntários, com 68 ± 5kg e 1,73 m, apresentavam-se eutróficos (IMC=23 ± 1) e clinicamente saudáveis, tendo-se constatado um consumo usual de dieta hiperproteica e hiperlipídica, para a maioria do grupo. A análise direta das dietas consumidas revelou que foi atendida 95% da recomendação de energia para o grupo (2809 ± 198,54 kcal), distribuída em 16, 26 e 58% para proteínas, lipídios e carboidratos, respectivamente. Ao contrário do Ca (712 ± 79 mg), as concentrações de Zn (11,62 ± 1,55mg) e Fe (10,65 ± 1,47mg) superaram as recomendações, e as frações IP5 e IP6 do fitato não foram detectáveis (<0,01). Das análises das concentrações de zinco no plasma, eritrócitos e urina, conclui-se que o estado nutricional, relativo ao zinco, da maioria dos voluntários mostrou-se satisfatório, uma vez estar de acordo com os padrões de referência. O enriquecimento dos isótopos 67Zn e 70Zn na urina de cada indivíduo foi determinado pela técnica de espectrometria de massa por plasma induzido (ICP-MS), revelando percentual de absorção de 30%, com variações entre 11 e 47%. Os resultados indicam que a biodisponibilidade do zinco na dieta avaliada pode ser classificada como sendo de moderada a alta, segundo critérios da OMS. / This research aimed to evaluate the bioavailability of zine in the Brazilian diet, containing the required energy and macronutrients, by the use of a stable isotope technique. Twelve male eutrophic healthy individuals in age between 19 - 42 years were enrolled in the study. The group consumed an experimental diet for a week, corresponding to 95% of the energy requirement (RDA-1989) and composed of 16% protein, 26% fat and 58% carbohydrate. In day eighth of the experiment, an intravenous injection of 70Zn was given to each individual and the meal consumed at lunch was extrinsically labeled with 67Zn, in that both stable isotopes were enriched. The percentage increase, or enrichment, of 67Zn and 70Zn in the urine of each individual was evaluated by Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and showed an average zinc absorption of 30%, variation from 11 to 47%, and a bioavailability from modest to high, according to the WHO criteria.

No description available.

Ο Ευρωπαϊκός Θαλασσοκόρακας (Phalacrocorax aristotelis) περιλαμβάνει τρία υποείδη που διαχωρίζονται με βάση μορφολογικές και συμπεριφορικές διαφορές και εξαπλώνονται σε διαφορετικές γεωγραφικές περιοχές. Το ατλαντικό υποείδος, P. a. aristotelis, κατά την αναπαραγωγική του περίοδο, εξαπλώνεται στον Ατλαντικό από τη Β. Ρωσσία μέχρι τις ατλαντικές ακτές της Ιβηρικής χερσονήσου, το υποείδος P. a. riggenbachi διαβιεί στις ακτές της Β. Αφρικής και το μεσογειακό (Phalacrocorax aristotelis desmarestii) θεωρείται ενδημικό υποείδος της Μεσογείου και της Μαύρης Θάλασσας. Η βιολογία και η οικολογία του μεσογειακού θαλασσοκόρακα δεν είναι μελετημένη, ιδιαίτερα στις ανατολικές περιοχές της εξάπλωσής του, παρότι η περιοχή του βόρειου Αιγαίου περιλαμβάνεται στις σημαντικότερες περιοχές αναπαραγωγής του είδους. Η παρούσα διδακτορική διατριβή, αποτελεί την πρώτη μελέτη σχετικά με την οικολογία της αναπαραγωγής και την ανάλυση της γενετικής δομής του μεσογειακού θαλασσοκόρακα σε αποικίες του Αιγαίου. Συγκεκριμένα, μελετώνται τέσσερα θέματα της βιολογίας του υποείδους: (1) η αναπαραγωγική επιτυχία και οι πιθανοί περιβαλλοντικοί παράγοντες που ενδέχεται να την επηρεάζουν, (2) η αναλογία του φύλου των νεοσσών, (3) οι διατροφικές συνήθειες κατά την αναπαραγωγική περίοδο, και (4) η γενετική δομή αναπαραγωγικών πληθυσμών του Αιγαίου, καθώς και οι φυλογεωγραφικές σχέσεις μεταξύ τους και με άλλους μη ελληνικούς πληθυσμούς. / The European Shag (Phalacrocorax aristotelis) is currently divided in three subspecies based on plumage differences, non-overlapping distributions and phenology. The nominate subspecies, P. a. aristotelis, has a breeding distribution from northern Russia to the Atlantic coast of Iberia, P. a. riggenbachi is found along the northern African coasts and the Mediterranean Shag (Phalacrocorax aristotelis desmarestii) is considered an endemic subspecies of the Mediterranean and the Black Seas. The Mediterranean subspecies’ biology and ecology are poorly studied, especially in the eastern part of its distribution, despite the fact that North Aegean Sea (Greece) is considered one of the most important regions for its reproduction. This study presents the first results regarding the study of its reproduction ecology and genetic structure in colonies from the Aegean Sea region. Specifically, four aspects of the its biology are addressed here: (1) breeding success and the possible ecological factors that may affect it, (2) the sex ratio of fledglings, (3) feeding habits during reproduction, and (4) the genetic structure of breeding populations in the Aegean and their phylogeographic relationships with other non-Greek populations.

This thesis was written within the framework of a biomanipulation project where young-of-the-year (YOY) pikeperch (Sander lucioperca) were stocked to a Baltic Sea bay to improve water quality through a top-down trophic cascade. The aim of my doctorial studies was however focused on a broader ecological question, namely predation (the main driving force in a biomanipulation). Hence, this thesis consists of four papers where we study the interactions between predator and prey using fish and zooplankton and how these interactions can be measured. In paper I we evaluated the performance of different diet analysis methods by individual based modelling and found that when having a nutritional gain perspective, mass based methods described diets best. Paper II investigated how the explorative, foraging and anti-predator behaviour of the YOY pikeperch used for stocking were affected by their rearing environment (pond vs. tank rearing). The more complex and varied environment in the semi-natural ponds seemed to promote a more flexible and active behaviour, better equipping young fish for survival in the wild. For paper III we studied the diel vertical migration in the six copepodite stages of the zooplankton Acartia spp. and Eurytemora affinis in relation to fish biomass, phytoplankton abundance and temperature. Both species migrated and in addition showed increased migration range with size within species, indicating evasion from visual predators. Paper IV addressed the movement of littoral Eurasian perch (Perca fluviatilis) via stable isotope signatures (13C and 15N) and body condition. We found clear indications of sedentarity and intra-habitat dietary differences. Interactions between predators and prey are complex and affected by both physiological and environmental characteristics as well as behavioural traits. The results in this thesis suggest that different species and even different life stages pursue different strategies to survive. / At the time of doctoral defense, the following papers were unpublished and had a status as follows: Paper 1: Submitted.  Paper 2: In press.  Paper 4: Submitted.