Abstract We examined the effects of different live food¡Bwater quality and environmental factor on the survival rate and growth of newborn Hippocampus kuda, in the laboratory, Three experiments were conducted: (1) effects of food organisms using three species i.e., Schmackeria dubia¡BArtemia sp. and Brachionus sp.; (2) effects of adding algae water to aquaria, and(3) effects of side-illumination and habitat. The survival rate and growth rate, were used to assess the responses of seahorses. The results showed that the growth of seahorses fed with copepod is the best, 28 days old seahorses are 127 ¡Ó 8mg (mean of three replicates ¡Ó 1se) in wet weight¡B32.38 ¡Ó 0.79mm in body length and the survival rate is 92.0 ¡Ó 0%. Copepod + artemia group are 69 ¡Ó 5mg¡B27.23 ¡Ó 0.35mm and 90.7 ¡Ó 5.8%. Rotifer + Copepod group are 86 ¡Ó 8mg¡B28.36 ¡Ó 1.04mm and 88.0 ¡Ó 4.0%. Rotifer + artemia group are 26 ¡Ó 1mg¡B19.77 ¡Ó 0.31mm and 68.0 ¡Ó 0%. Artemia group are 45 ¡Ó 2mg¡B23.29 ¡Ó 0.41mm and 38.7 ¡Ó 3.5%. The lowest survival rate occurs in the rotifer group and the no-feed group, all seahorses were dead on the sixteenth day and the thirteenth day, respectively. Copepods were used as food organisms in side-illumination ¡Balgae and habitat experimental designs. The difference were all signify- cant between the side-illumination and no side-illumination group¡G97.3 ¡Ó 1.3%¡B68.0 ¡Ó 6.9%( p=0.014, ANOVA, Fisher¡¦s PLSD)¡A126 ¡Ó 3mg¡B101 ¡Ó 5mg(p=0.013) and 32.51 ¡Ó 0.30mm¡B29.32 ¡Ó 0.75mm(p=0.017)¡C The body length was significant difference between algae fed and no-algae group: 25.40 ¡Ó 0.63mm¡B23.23 ¡Ó 0.44 mm(p= 0.047). The survival rate¡Bbody weight and body length were all no significant difference between habitat and no habitat group. To sum up, the living food is a major factor affecting survival and growth. And the side-illumination method can increase both. So the suitable living food and aquaculture engineer designs have great potential to raise more seahorses.

A 12-year retrospective study of 172 deceased captive syngnathids (Hippcampus kuda, H. abdominalis, and Phyllopteryx teaniolatus) from the Toronto Zoo was performed. The most common cause of mortality was an ulcerative dermatitis, occurring mainly in H. kuda. The dermatitis often presented clinically as ‘red-tail’, or hyperaemia of the ventral aspect of the tail caudal to the vent, or as multifocal epidermal ulcerations occurring anywhere. Light microscopy often demonstrated filamentous bacteria associated with these lesions, and it was hypothesized that the filamentous bacteria were from the Flavobacteriaceae family. Bacteria cultured from ulcerative lesions and DNA extracted from ulcerated tissues were examined using universal bacterial 16S rRNA gene primers. A filamentous bacterial isolate and DNA sequences with high sequence identity to Cellulophaga fucicola were obtained from ulcerated tissues. Additionally, in situ hybridization using species-specific RNA probes labeled filamentous bacteria invading musculature at ulcerative skin lesions.

No description available.

Academic: Ovarian cancer is the leading cause of death among female gynecologic cancers. Current treatments include surgical debulking, and chemotherapy. However, better interventions are needed to reduce the mortality rate of metastatic disease. Ovarian cancer cells have displayed the ability to aggregate and form 3D homogeneous and heterogeneous spheroids, which can function as micrometastases. Ovarian cancer spheroids survive independently prior to adhering to an endothelial tissue. Since aggregation has been shown to provide a survival advantage to the spheroids and increased their aggressive phenotype, this study aimed to investigate how the metabolism of ovarian cancer cells change in 3-dimensional (3D) culture. Examining metabolic pathways and identifying markers of metabolic change could provide the scientific base for new, targeted interventions for this disease. Spheroids of both homogeneous and heterogeneous composition demonstrated overall lower metabolic capacity than their adherent counterparts. Spheroids had a lower basal energetic demand than adherent cells, paralleled by lower maximal respiration capacity, glycolytic capacity, and spare respiratory capacity. We conclude that the lower energetic demand of spheroids may be a mechanism to prolong death by reserving energy and metabolic cellular processes; this may render anti-metabolic drug treatment with AICAR or metformin ineffective against disseminating ovarian cancer aggregates. General: Ovarian cancer is currently the leading cause of death among female gynecologic cancers. While treatments exist, better interventions are needed to reduce the mortality rate in this form of cancer. Ovarian cancer cells have displayed the ability to aggregate and form 3D homogeneous and heterogeneous spheroids, which can function as micrometastases. Ovarian cancer spheroids survive independently prior to adhering to an endothelial tissue. Since aggregation has been shown to provide a survival advantage to the spheroids and increased their aggressive phenotype, this study aims to investigate how the metabolism of ovarian cancer cells change in 3-dimensional (3D) culture. Examining metabolic pathways and identifying markers of metabolic change could provide the scientific base for new, targeted interventions for this disease. / Master of Science

Seahorses, genus Hippocampus erectus, are subject to large and continuously-growing international trade. Concerns with the effects of trade on seahorse population worldwide have led to their international protection by the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). In order to manage seahorse populations, we first need to understand their basic biology. The purpose of this project was to establish an energy budget for Hippocampus erectus from Tampa Bay, Florida. A total of 108 specimens were collected throughout Tampa Bay from 2004 to 2007 using 21.3 m offshore seines and 6.1 m otter trawls. Those specimens were utilized to determine different components of the energy budget: size at maturity, feeding and nutrition within a captive environment, and metabolic rate. Seahorses collected in the study ranged in size from 4 mm to 160 mm, and were 0 to 4 years in age. Conventional methods of aging fish could not be applied to this species. Instead, an alternative method that involved sectioning vertebrae and using Scanning Electron Microscopy (SEM) to enumerate age-specific marks on vertebra was employed. Age data were then paired against length-weight charts to estimate age by length and mass. Length-frequencies were segregated by sex and compared to the growth data. No sex-specific differences were found. Fish reared in captivity were also used for nutrition, respiration, excretion analysis. Hippocampus erectus in captivity were calculated to have a resting respiration rate of 85.9 microliter O2/g/h and an excretion rate of 0.48 mmol NH3/g/h. The viscera of H. erectus and Artemia spp. were broken into protein and lipid content by caloric composition. The overall percentage of protein per seahorse was 11% and 17% by lipid concentration. Overall the sum of energy cost of H. erectus accounted for 81% of the total energy ingested.

Brain size varies greatly among vertebrates. It has been proposed that the diversity of brain size is produced and maintained through a balance of adaptations to different types and levels of cognitive ability and constraints for adaptive evolution. Phylogenetic comparative studies have made major contributions to our understanding of brain size evolution. However, previous studies have nearly exclusively focused on mammalian and avian taxa and almost no attempts have been made to investigate brain size evolution in ectothermic vertebrates. In my thesis, I studied brain size evolution in two groups of fish with extreme diversity in ecology, morphology and life history, Cichlidae and Syngnathidae. Using phylogenetic comparative methods, I investigated four key questions in vertebrate brain size evolution; cognitive adaptation, sexual selection, phenotypic integration and energetic constraints. I have demonstrated i) that phenotypic integration can link functionally unrelated traits, and this may constrain independent evolution of each part involved or promote concerted evolution of an integrated whole, ii) that brain-body static allometry constrains the direction of brain size evolution, even though the static-allometry showed ability to evolve, allowing evolution of relative brain size under allometric constraints, iii) that the energetic constraints of development and maintenance of brain tissue is an important factor in forming the diversity in brain size in cichlids and syngnathids, both at macroevolutionary and microevolutionary time scales, and iv) that adaptation for feeding and female mating competition may have played key roles in the adaptive evolution of brain size in pipefishes and seahorses. To conclude, my thesis shows the strong benefit of using fish as a model system to study brain size evolution with a phylogenetic comparative framework.

This work comprehensively reviews the international trade in sea horses, particularly the Hippocampus genus, from 1997 to 2012. The genus was added to the Appendices II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (thereinafter as the CITES) in 2002, came into force in 2004. Before the addition, the genus was entered into the appendix D for the non-CITES species whose import is monitored by the EU. The CITES Trade Database kept by the UNEP - WCMC Secretariat was used as the source of data about the trade. The frequency of the trade in sea horses tended to be increasing with a moderate decrease in 2009. The Hippocampus species was traded in the following forms: live, bodies, derivatives, specimens, medicine, carvings, eggs live, extracts, fingerlings, powder, skeletons, skin pieces, soup, trophies and other unspecified forms. The living specimens traded: (1) specimens taken from the wild., (2) animals bred in captivity as well as parts and derivatives, (3) Animals born in captivity (F1 or subsequent generations) that do not fulfil the definition of 'bred in captivity', as well as parts and derivatives thereof, (4) specimens of animals reared in a controlled environment, taken as eggs or juveniles from the wild, where they would otherwise have had a very low probability of surviving to adulthood, (5) confiscated or seized specimens, (6) pre- Convention specimens. The trades purposes were: (1) commercial, (2) personal, (3) scientific, (4) zoo (aquarium) breeding, (5) educational, (6) breeding in captivity, circus and other exhibitions, (7) conventional medicine, (8) law enforcement / judicial / forensic purposes. The most traded genus was unspecified (Hippocampus spp.), then the H. kuda, the H. reidi and the H. erectus as living specimens or dead bodies; also the ones from the wild and for the commercial purposes. Those came mostly from Vietnam, Thailand or Australia. They were exported mainly from Indonesia, Sri Lanka and Australia. The greatest importers were the USA, Germany and the Great Britain.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-02-21T12:32:04Z No. of bitstreams: 1 arquivo total.pdf: 2615481 bytes, checksum: a12febbc7b66acbd174fcad4f96d07cd (MD5) / Made available in DSpace on 2017-02-21T12:32:04Z (GMT). No. of bitstreams: 1 arquivo total.pdf: 2615481 bytes, checksum: a12febbc7b66acbd174fcad4f96d07cd (MD5) Previous issue date: 2015-09-23 / A utilização do peróxido de hidrogênio (H2O2) como forma de prevenção e controle de mortalidades associadas com infecções fúngicas, bacterianas, e de infestações parasitárias em diferentes espécies e ciclos de vida dos peixes é comum na aquicultura. Sabe-se que o H2O2 afeta a atividade de transcrição celular da base-redox, o controle a respostas inflamatórias e as respostas imunológicas, estando diretamente relacionados ao estresse. A grande vantagem do H2O2 é o impacto ambiental mínimo relacionado com a ausência de resíduos tóxicos; nesse sentido, estudos direcionados ao desenvolvimento de protocolos que se utilizem de produtos de baixo risco ambiental para o tratamento de patologias se fazem necessários. Adicionalmente, estudos que visam ampliar os conhecimentos etológicos como um método não-invasivo de avaliação do bem-estar animal são de grande importância na avaliação do estresse. Neste trabalho, foi realizado um estudo com Hippocampus reidi, visando ampliar o conhecimento a respeito do seu comportamento em condições normais e de estresse, bem como avaliar o impacto e implicações do uso de H2O2 no manejo/cultivo de cavalos-marinhos e possível forma de remediação/mitigação desses efeitos. Foi utilizado um total de 43 espécimes de H. reidi, provenientes da população estoque para cultivo experimental do LAPEC (Laboratório de Peixes – Ecologia e Conservação), Universidade Federal da Paraíba, Brasil. Observou-se que a presença de animais parece ser um fator decisivo na degradação do H2O2, por acréscimo de matéria orgânica e bactérias ao sistema, como também pela própria absorção por parte dos animais. Foi constatado ainda que o H2O2 tem o potencial para ser usado em tratamento por imersão (banho) em H. reidi, tendo em vista que concentrações abaixo de 545 μM mostraram-se seguras em um período de 60 min de exposição para a espécie. Por outro lado, o peróxido de hidrogênio é tóxico em concentrações igual/superior a 599 μM e teve profundo impacto sobre o peso, comportamento e frequência respiratória em H. reidi. Foi testado também se Mn-porfirina MnTE-2-PyP5+ (MnP) é um potencial antioxidante catalítico para manutenção e cultivo de H. reidi, minimizando os impactos causados por tratamentos de altas concentrações de H2O2. Os resultados iniciais obtidos mostraram que o MnP foi benéfico a H. reidi, uma vez que minimizou os impactos causados por tratamentos de altas concentrações de H2O2 a longo prazo, apresentando assim um amplo potencial a ser estudado nessa área. / A utilização do peróxido de hidrogênio (H2O2) como forma de prevenção e controle de mortalidades associadas com infecções fúngicas, bacterianas, e de infestações parasitárias em diferentes espécies e ciclos de vida dos peixes é comum na aquicultura. Sabe-se que o H2O2 afeta a atividade de transcrição celular da base-redox, o controle a respostas inflamatórias e as respostas imunológicas, estando diretamente relacionados ao estresse. A grande vantagem do H2O2 é o impacto ambiental mínimo relacionado com a ausência de resíduos tóxicos; nesse sentido, estudos direcionados ao desenvolvimento de protocolos que se utilizem de produtos de baixo risco ambiental para o tratamento de patologias se fazem necessários. Adicionalmente, estudos que visam ampliar os conhecimentos etológicos como um método não-invasivo de avaliação do bem-estar animal são de grande importância na avaliação do estresse. Neste trabalho, foi realizado um estudo com Hippocampus reidi, visando ampliar o conhecimento a respeito do seu comportamento em condições normais e de estresse, bem como avaliar o impacto e implicações do uso de H2O2 no manejo/cultivo de cavalos-marinhos e possível forma de remediação/mitigação desses efeitos. Foi utilizado um total de 43 espécimes de H. reidi, provenientes da população estoque para cultivo experimental do LAPEC (Laboratório de Peixes – Ecologia e Conservação), Universidade Federal da Paraíba, Brasil. Observou-se que a presença de animais parece ser um fator decisivo na degradação do H2O2, por acréscimo de matéria orgânica e bactérias ao sistema, como também pela própria absorção por parte dos animais. Foi constatado ainda que o H2O2 tem o potencial para ser usado em tratamento por imersão (banho) em H. reidi, tendo em vista que concentrações abaixo de 545 μM mostraram-se seguras em um período de 60 min de exposição para a espécie. Por outro lado, o peróxido de hidrogênio é tóxico em concentrações igual/superior a 599 μM e teve profundo impacto sobre o peso, comportamento e frequência respiratória em H. reidi. Foi testado também se Mn-porfirina MnTE-2-PyP5+ (MnP) é um potencial antioxidante catalítico para manutenção e cultivo de H. reidi, minimizando os impactos causados por tratamentos de altas concentrações de H2O2. Os resultados iniciais obtidos mostraram que o MnP foi benéfico a H. reidi, uma vez que minimizou os impactos causados por tratamentos de altas concentrações de H2O2 a longo prazo, apresentando assim um amplo potencial a ser estudado nessa área.

Indiana University-Purdue University Indianapolis (IUPUI) / Human immunodeficiency virus (HIV) primarily infects T helper (Th) cells. Decrease in the number of Th cells is the hallmark of HIV infection. Latent reservoirs of human immunodeficiency virus (HIV) are the leading barrier towards eradication of HIV infection. T Follicular helper (Tfh) cells are a subset of Th cells that function to provide aid to B cells for their maturation, affinity selection and antibody class switch. Several studies have shown that Tfh cells are a major reservoir of latent as well as productive hiv infection. But in contrast to the fate of other Th cell subsets, the frequency of Tfh cells was shown to have increased during HIV infection which could not be attributed to their reduced susceptibility to HIV infection. The hypothesis was that Tfh cells possess a unique metabolic phenotype that protects them from HIV induced cell death. Transcriptome analysis of Th subsets from human donors and showed that Tfh cells rely less on glycolysis for their energetic requirements and instead have increased transcription of fatty acid synthesis genes. This finding was corroborated by seahorse extracellular flux assay. The results shoId that glycolysis was not essential for Tfh cell differentiation in-vitro. The observed increase in Tfh cell frequency could not be attributed to increased Tfh differentiation upon HIV infection since HIV infection inhibited the differentiation of both non-Tfh and Tfh cells. The results found that bypassing the glycolytic pathway by providing Tfh cells with Galactose in the medium protected ex-vivo infected primary tonsillar cells from HIV induced cell death. This protection could be partly explained by the induction of Baculovirus IAP repeat containing 5 (BIRC5) when the cells utilized Galactose instead of Glucose. The studies together show that Tfh cells have an oxidative metabolic phenotype which protects them from HIV induced cell death in part by induction of BIRC5 expression.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第24487号 / 医博第4929号 / 新制||医||1063(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 竹内 理, 教授 上野 英樹, 教授 髙折 晃史 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM