Dissertação(mestrado) - Universidade Federal do Rio Grande, Programa de Pós–Graduação em Oceanografia Biológica, Instituto de Oceanografia, 2006. / Submitted by Cristiane Gomides (cristiane_gomides@hotmail.com) on 2013-11-19T10:53:47Z No. of bitstreams: 1 Dissertacao Fernanda Caldeira.pdf: 2786616 bytes, checksum: 7d8912a4eb7053a789229e293eca615b (MD5) / Approved for entry into archive by Angelica Miranda (angelicacdm@gmail.com) on 2013-11-20T21:32:49Z (GMT) No. of bitstreams: 1 Dissertacao Fernanda Caldeira.pdf: 2786616 bytes, checksum: 7d8912a4eb7053a789229e293eca615b (MD5) / Made available in DSpace on 2013-11-20T21:32:49Z (GMT). No. of bitstreams: 1 Dissertacao Fernanda Caldeira.pdf: 2786616 bytes, checksum: 7d8912a4eb7053a789229e293eca615b (MD5) Previous issue date: 2006 / O desenvolvimento da raia Sympterygia acuta foi descrito baseado em uma série completa de embriões do estágio 17, o primeiro estágio onde a forma do corpo e a primeira fenda branquial podem ser distinguidos, até a eclosão. Os embriões foram provenientes de cápsulas ovígeras coletadas na praia de Cassino, estado do Rio Grande do Sul, Brasil. As características externas destes espécimes foram usadas para estabelecer uma tabela de estágios de S. acuta. Cada estágio é definido por um conjunto de características morfológicas que inclui a abertura e a posição das sucessivas fendas branquiais, do tamanho e da forma das nadadeiras peitorais (disco) e pélvicas, da forma da boca e da posição dos espiráculos. A relação entre a massa de vitelo e o crescimento do embrião em massa está de acordo com o padrão conhecido para elasmobrânquios ovíparos e vivíparos lecitotróficos. O Estágio 30 é marcado por grandes mudanças, como a pré-eclosão, o início da entrada de vitelo no sistema digestivo do embrião, e início do real crescimento embrionário. O estudo fornece uma comparação com outros embriões de Chondrichthyes baseada nas características externas. O desenvolvimento das nadadeiras peitorais, que desde o começo ocupam toda a face lateral da região abdominal, é uma divergência importante entre esta raia e tubarões, exceto Squatinidae. Isto pode ser uma evidência de que filogenéticamente as Rajidae e Squatinidae são próximas, e que o desenvolvimento embrionário aporta informações relevantes para o estudo da filogenia. / The development of the skate Sympterygia acuta is describe based on a complete series of embryos from stage 17, the earliest stage in which the body forms and first visceral cleft can be distinguished, to hatching. The embryos were obtained from egg-cases caught at Cassino Beach, State of Rio Grande do Sul, Brasil. The external features of these specimens are used to establish a stage table of S. acuta. Each stage is defined by a suite of morphological characters including the opening and position of the successive visceral clefts, size and shape of pectoral (disc) and pelvic fins, shape of mouth and position of spiracles. The yolk-embryo weight relationship agrees with the pattern found in other elasmobranchs oviparous and viviparous lecithotrophic. The stage 30 is marked by great changes, as the pre-hatching, the beginning of the entrance of yolk in the digestive system of the embryo, and the beginning of the real embryonic growth. The study provides a comparison with other condrichthyan embryos based on external features. The development of the pectoral fins, which since the beginning occupies the whole side of the abdominal region, is the most important divergence between this skate and the sharks, except from Squatinidae. This might be an evidence that Rajidae and Squatinidae have a close phylogenetic relationship.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-10-13T11:36:18Z No. of bitstreams: 1 julianapolisseni.pdf: 645848 bytes, checksum: 5362a586a32461dda31a0e7f5d2e9fef (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-10-22T12:55:07Z (GMT) No. of bitstreams: 1 julianapolisseni.pdf: 645848 bytes, checksum: 5362a586a32461dda31a0e7f5d2e9fef (MD5) / Made available in DSpace on 2016-10-22T12:55:07Z (GMT). No. of bitstreams: 1 julianapolisseni.pdf: 645848 bytes, checksum: 5362a586a32461dda31a0e7f5d2e9fef (MD5) Previous issue date: 2008-08-29 / O impacto da técnica de biópsia sobre o desenvolvimento embrionário posterior ainda foi pouco estudado e também não foi estabelecido um protocolo único e universal para o PGD. Outro ponto a considerar é a pequena quantidade de DNA genômico disponível para se realizar os estudos genéticos, pelo número reduzido de células obtidas pela técnica de biópsia. Metodologias que utilizam whole genome amplification (WGA) vêm sendo desenvolvidas. Utilizando este método é possível gerar microgramas de DNA partindo de pequenas quantidades de DNA. Então, o objetivo deste estudo foi avaliar o desenvolvimento de embriões bovinos submetidos à biópsia nos estádios de 8-16 células e o uso da técnica de amplificação de todo o genoma nos blastômeros retirados, para posterior identificação do sexo através do PCR. Um grupo de 706 complexos cumulus-ovócitos (CCOs) de bovinos foram maturados e fertilizados in vitro, em estufa incubadora a 38,8 °C, 95% de umidade e 5% de CO2. Os prováveis zigotos foram semi-desnudados e cultivados no meio CR2aa sob as mesmas condições da fertilização. No quarto dia após a fertilização, embriões bovinos com 8-16 células foram aleatoriamnte distribuídos entre dois grupos: controle (n=103) e biópsia (n=92). O número de células removidas correspondeu à quarta parte do embrião. Os blastômeros retirados foram submetidos à amplificação de todo o genoma seguido por PCR. Os embriões retornaram ao meio de cultivo para avaliação do desenvolvimento embrionário. Avaliou-se pelo teste do qui-quadrado a produção de blastocisto no oitavo dia de cultivo, a taxa de eclosão no décimo dia de cultivo, a eficiência da amplificação de todo o genoma e da identificação do sexo nos blastômeros removidos. O número de células embrionárias foi analisado por análise de variância ANOVA (Instituto SAS, Inc., Cary, NC, USA). A proporção de blastocisto no oitavo dia de cultivo foi avaliada pelo teste de Wilcoxon. Diferenças foram consideradas significantes se P<0,05. Um total de 92 embriões bovinos foi utilizado para realização da técnica de biópsia e apresentaram produção de blastocisto de 53,3%, com 44,9% de taxa de eclosão. Essas taxas foram semelhantes (P>0,05) às dos 103 embriões do grupo controle (66,0% e 42,6%, respectivamente). Não houve variação no número de células embrionárias entre os grupos e também não ocorreu diferença na proporção de blastocisto entre os grupos controle e biópsia no oitavo dia de cultivo (P>0,05). Os blastômeros retirados foram submetidos à amplificação de todo o genoma, com 98,2% de eficiência. Entretanto, só foi possível identificar o sexo em 59% das amostras. Conclui-se que a técnica de biópsia realizada em embriões bovinos de 816 células não afetou o desenvolvimento embrionário subseqüente nem a qualidade embrionária dos embriões. A utilização do kit de amplificação de todo o genoma mostrou-se eficiente nos blastômeros retirados, sendo possível identificar o sexo em mais da metade dos embriões. . / The impact of biopsy technique on posterior embryo development is still poorly studied and there is no single universally practiced protocol for implementing PGD. Another thing to consider is the small amount of genomic DNA available to perform the genetic study, due to the reduced number of cells obtained from the biopsy. Alternatively, methodologies using whole genome amplification (WGA) have been developed. Using these methods, it is possible to generate microgram quantities of DNA starting with a little genomic DNA. The aim of this study was to evaluate the effect of biopsy at 8-to 16-cell bovine embryos on subsequent development and WGA on removed blastomeres. A group of 706 complexes cumulus oocytes (CCOs) obtained from local slaughterhouse ovaries were matured and fertilized in vitro, in incubator at 38.8°C with 95% humidified air and 5% of CO2. The zygotes were semidenuded and cultured in CR2aa medium under the same conditions as with in vitro fertilization. On the fourth day after fertilization the 8-to 16-cell embryos were distributed randomly across two groups: control (n=103) and biopsy (n=92). The amount of cells removed cells was one-fourth the embryo of blastomeres. The removed blastomeres were submitted to whole genome amplification (WGA) followed by PCR. The embryos were returned to culture for development evaluation. The chisquare test was used for statistic evaluation of the results of blastocyst rate on the eight day after fertilization and hatching rate on tenth day between biopsy and control group, to test the WGA efficiency and to determine the sex proportion of biopsied samples submitted to PCR. The number of cell per embryo was analyzed by ANOVA with SAS (SAS Institute, Inc., Cary, NC, USA). The blastocyst proportion was evaluated with Wilcoxon test. Differences were considered significant if P<0.05. A total of 92 embryos were submitted to biopsy technique. The blastocyst production was 53.3%, with 44.9% of hatching rate. This rate was similar to control group (66.0% and 42.6%) found on 103 embryos. Overall, no impact was detected on embryo quality in blastocyst cell number between the two groups. The proportion of blastocyst in different stages of development on the 8th day did not differ among groups (P>0.05). Removed blastomeres were submitted to WGA, resulting in 98.2% of efficiency. However, only 59% of samples were sexed by PCR. In conclusion biopsy of 8-to 16-cell bovine embryos did not affect subsequent development. WGA was successful in removed blastomeres and was possible to determinate the sex in the samples.

Hurricanes can cause considerable damage along the coastlines of the United States. There are very little data relating hurricanes and sea turtle nesting. Hurricane season (June- November) and sea turtle nesting season, for the study period, (March- September) overlap in Broward County, Florida. This paper examines the relationship between the Hurricane Impact Index (HII) and hatching success of loggerhead sea turtles (Caretta caretta) and HII and washout rate. For ten of the eleven storms studied, there was no significant relationship between HII and hatching success (percent). Tropical Storm Ophelia (2005) showed a significantly higher (p=0.0206) hatching success after the storm’s passing than before the storm’s passing. There was a significant relationship between washout rate and HII (p=0.026).

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21967号 / 農博第2357号 / 新制||農||1070(附属図書館) / 学位論文||R1||N5218(農学部図書室) / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 近藤 直, 准教授 小川 雄一, 教授 清水 浩 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DGAM

MSCAGR (Animal Science) / Department of Animal Science / The use of in vitro culture media and assisted hatching techniques remain a challenging obstacle to hatching of blastocyst-stage embryos. Mechanical, chemical, enzymatic thinning and laser assisted techniques have been used previously, but there is still a lack of information on its application and implication in livestock. The aim of this study was to compare the effect of two in vitro culture media ((Ham’s F10 and Tissue Culture Medium 199 (TCM-199)) and four assisted hatching techniques (mechanical, chemical, enzymatic and laser) on blastocyst formation and hatching rate using murine embryos as a model. The C57BL/6 and BALB/c mouse breeds were bred and raised until they reach maturity and then bred naturally to produce a hybrid F1 generation. The light in the breeder house was controlled at 14 hours light and 10 hours darkness. Feed and water were provided ad libitum for the mice. Mature female mice were super-ovulated using equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG). A total of 400 blastocysts were collected from the F1 generation and these were allocated equally for the four assisted hatching techniques (laser, mechanical, chemical and enzymatic) as well as a non-treated control group. The blastocysts were paired into a group of 10 and replicated 4-four times for each assisted hatching techniques and control group. The embryos were then cultured for 24 hours and the hatching of the embryos were observed. Hatched embryos were stained for blastomere counting. The general linear model (GLM) of statistical analysis software (SAS) version 9.4 was used to analyze the data. Assisted hatching techniques (laser, mechanical, enzymatic and chemical) yielded 46.86±37.12; 51.07±40.19; 39.05±35.83 and 33.32±37.50% of hatching, respectively under in vitro culture in Ham’s F10. There was a significant difference (p<0.05) observed between assisted hatching techniques using Ham’s F10 as culture medium. In the TCM-199, laser, mechanical, enzymatic and chemical assisted hatching techniques yielded 56.25±43.30; 52.55±35.50; 49.16±37.50 and 33.85±35.50%, respectively, with significant differences (p<0.05). However, the hatching rate of embryos for all techniques was higher when in vitro cultured in TCM-199 compared to those cultured in Ham’s F10, and statistically higher than the control group. In conclusion, laser assisted hatching technique is the best of the techniques to use to assist the hatching of murine embryos and TCM-199 is the best of the two in vitro culture media for the hatching percentage.

In der Archäologie ist das Zeichnen von Fundobjekten ein zeitaufwändiger und daher kostenintensiver Prozess. Die Abbildungen sind wegen fehlender Standards sowie den unterschiedlichen persönlichen Stilen und Fähigkeiten der Zeichner subjektiv und stark inhomogen. Es gibt deswegen erste Ansätze, mithilfe von 3D-Scannern virtuelle Modelle zu erzeugen, von denen dann objektive, standardisierte Abbildungen extrahiert werden sollen. Von den zahlreichen Verfahren aus dem Bereich des Non-Photorealistic Rendering (NPR) kommen dafür insbesondere solche in Betracht, die Verzierungen und Oberflächeneigenschaften betonen. Auf Interaktion soll dabei nach Möglichkeit verzichtet werden.

Diapause is often an adaptation for survival during periods of harsh environmental conditions. Some diapausing stages do not terminate the dormancy once the favourable conditions are restored. Such prolonged diapause may be enforced by environment if a diapausing stage cannot be reached by the cues inducing termination of dormancy. However, it may also be an advantageous bet-hedging strategy to allow only a fraction of dormant stages produced in any given season to hatch the next time conditions become favourable. I tested whether such strategy can be observed in hatching patterns of dormant eggs of Daphnia obtusa - a cladoceran occurring in small Central European temporary waters. I investigated the influence of intensity of illumination on hatching success, and effect of isolating the eggs encased in ephippia from the sediment. Fraction of eggs terminating diapause, fraction of embryos successfully leaving the egg membranes, and timing of the response were assessed at 15 ˚C under four intensities of illumination (100% = 35µmol.m2 .s-1 , 75%, 50%, 25%; photoperiod 12h light: 12h dark) and in complete darkness for 21 days. My results support previous suggestions that there is no genetically-fixed bet-hedging strategy in D. obtusa. I observed high proportion of eggs which terminated diapause in all...

The spawning time differs between sympatric ecotypes of lake-dwelling whitefish (Coregonus lavaretus) and this temporal divergence can be linked to the type of spawning habitat used (i.e., streams, littoral or profundal). It has been hypothesized that the divergence in spawning time between spawning habitats is due to differences in incubation temperature regimes, and a need to synchronize hatching with the spring zooplankton bloom. An alternative hypothesis is that the hatching time of eggs is controlled by cues such as rising temperatures or increasing day length, and the divergence in spawning time occurs due to later maturation in colder habitats. To test these hypotheses, whitefish eggs were sampled on different spawning grounds and the median hatching time was estimated using logistic regression. Temperature logger data and information based on interviews and earlier studies were used to study accumulation of thermal energy during incubation in the form of degree-days. According to this thesis, the hypothesis that hatching time is dependent on rising temperatures and day length is supported, due to high variation in the amount of accumulated thermal energy, and little variation in hatching time. The egg sampling was not successful on profundal spawning grounds, nor on littoral spawning grounds of old populations, which made the results of this study weaker. Further studies are needed to fully confirm either of these hypotheses regarding spawning time of mature whitefish and hatching time of eggs. Sampling methods need to be advanced in order to enable egg sampling on all types of spawning grounds.

https://dc.etsu.edu/entc-2160-oer/1023/thumbnail.jpg

Habitat loss is among the biggest threats to conservation worldwide, so habitat restoration plays an increasing role in endangered species management. This is especially true for species with high site fidelity, such as nesting marine turtles. Sand replenishment is commonly used to restore coastal beaches after severe erosion events, and may affect marine turtles and other species that live or reproduce in that habitat. I investigated how abiotic characteristics of sand used in a dune restoration project at Archie Carr National Wildlife Refuge, Florida, affected reproduction of the federally-endangered green turtle (Chelonia mydas). Sand structure and composition can affect egg development and hatching success by altering nest conditions, with nests in fine-grain or very coarse sand suffering decreased hatching success. I determined that calcium carbonate content (27.0% ± 1.4 SE vs. 15.1% ± 3.8 SE), moisture content (3.29% ± 0.26 SE vs. 4.59% ± 0.25 SE), and grain size (427.53 µm ± 14.1 SE vs. 274.66 µm ± 29.1 SE) differed significantly between natural and restored dunes. Hatching success of green turtles (44.7% ± 6.2 SE vs. 65.8% ± 5.3 SE) was significantly lower on restored dunes compared to natural dunes with an estimated loss of 22,646 hatched eggs. Hatching success also decreased as the nesting season progressed. These results demonstrate the importance of regulating fill material used in beach restoration projects; substrate characteristics are easily evaluated and can significantly influence marine turtle hatching success.