Made available in DSpace on 2016-05-02T13:55:47Z (GMT). No. of bitstreams: 1 TochimaraAparecidaMiyauchi-dissertacao.pdf: 1277501 bytes, checksum: 28f321c050df674dd0101fe282f265af (MD5) Previous issue date: 2012-09-28 / Coordenacao de Aperfeicoamento de Pessoal de Nïvel Superior / Parthenogenesis has been described as an alternative method to produce embryos for studies with embryonic cells, particularly in humans which have a restriction of fertilized embryos. Procedures used in parthenogenesis are also required to produce embryos by somatic cell nuclear transfer in domestic species. However, many biological, cellular and molecular aspects of parthenogenetic embryos are still unknown. This study aimed to compare the kinetics of development, apoptosis rate and gene expression of stress and celular metabolism in bovine parthenogenetic embryos and embryos fertilized in vitro. Oocytes (n = 1541) obtained from slaughterhouse ovaries were matured in vitro and submitted to parthenogenetic activation (4.62 µM ionomycin for 5 min followed by 2 mM 6-DMAP for 4 h) or in vitro fertilization (2 x 106 sperm/mL for 20h with semen from a single departure). 72h postactivation/fertilization (hpaf), embryos (8-cell) were frozen for subsequent analysis of gene expression and another part has been separated into groups of high or low potential of development: Part &#8805; 8 - parthenogenetic embryos with 8 or more cells (high development potential); Part < 8 - embryos less than 8 cells (low development potential); &#8805; 8 IVF - in vitro fertilized embryos with 8 or more cells; and IVF < 8 - embryos with less than 8 cells. Embryos were cultured in CR2aa medium with 2.5% BFS in 5% CO2, 5% O2, 90% N2 at 38.5° C and were evaluated rates of blastocyst 168hpaf (D7) and 196hpaf (D8). 8-cell embryos obtained after 72hpfa were analyzed for gene expression. D8 blastocysts were fixed and subsequently apoptotic index was analyzed by TUNEL. Data were compared by analysis of variance and means by Student Newman Keuls test. Gene expression was evaluated by REST® software. Values are shown as mean ± standard error. Embryos with 8 or more cells produced higher (P < 0.01) blastocysts rates in D7 and D8 compared to embryos with less than eight cells, showing its greatest development potential, regardless if they were parthenogenetic or fertilized. Embryos of Part &#8805; 8 group had higher (P <0.05) blastocyst rate in D7 (63.6 ± 3.4%) than IVF &#8805; 8 (45.3 ± 8.9%), but at D8 rate was similar (56.7 ± 3.0% and 44.2 ± 8.9% for Part &#8805; 8 and IVF &#8805; 8, respectively; P <0.05). There was no difference (P < 0.05) on blastocyst rates at D7 and D8 between embryos with less than 8 cells derived from parthenogenesis or fertilization. There was no difference in total cell number (92.0 ± 3.4, 102.30 ± 4.8), apoptotic cells (10.8 ± 1.22, 9.5 ± 1.07) and apoptotic index (11.4 ± 1.27, 9.8 ± 1.8) for parthenogenetic and IVF blastocysts, respectively. In 8-cell embryos analyzed, there was a subexpression of genes DNAJB1, HSPA1L, HSF1 and GLUT1 for Part in relation to FIV, while HSF2 was overexpressed in Part compared to FIV. In conclusion, bovine parthenogenetic embryos differ to embryos fertilized in vitro in hability of preimplantation development in vitro and gene expression, which may limit the use of parthenogenesis in studies of embryonic development. / A partenogênese tem sido descrita como um método alternativo para produzir embriões para estudos com células embrionárias, principalmente em humanos para os quais existe a restrição do uso de embriões fecundados. Procedimentos utilizados na partenogênese são também necessários para se produzir embriões por transferência nuclear com células somáticas nas espécies domésticas. Contudo, muitos aspectos biológicos, celulares e moleculares dos embriões partenogenéticos ainda são desconhecidos. Este estudo objetivou comparar a cinética do desenvolvimento, índice de apoptose e expressão de genes de estresse e metabolismo celular em embriões bovinos partenogenéticos e embriões fecundados in vitro. Oócitos (n=1541) obtidos de ovários de matadouro foram maturados in vitro e submetidos à ativação partenogenética (4,62 µM ionomicina por 5 min seguido de 2 mM 6-DMAP por 4h) ou fecundação in vitro (2 x 106 espermatozoides/ml por 20h, com sêmen de uma única partida). Com 72h pós-ativação/fecundação (hpaf) parte dos embriões (8 células) foram congelados para posterior análise da expressão gênica e outra parte foi separada em grupos de alto ou baixo potencial de desenvolvimento: Part&#8805;8 - embriões partenogenéticos com 8 ou mais células (alto potencial de desenvolvimento); Part<8 - embriões com menos de 8 células (baixo potencial de desenvolvimento); FIV&#8805;8 - embriões fecundados in vitro com 8 ou mais células; e FIV<8: embriões com menos de 8 células. Os embriões foram cultivados em meio CR2aa com 2,5% SFB em 5%CO2, 5%O2, 90% N2 a 38,5ºC e avaliadas as taxas de blastocistos com 168hpaf (D7) e 196hpaf (D8). Embriões com 8 células obtidos após 72hpfa foram analisados quanto à expressão gênica. Blastocistos em D8 foram fixados e posteriormente foram avaliados pela técnica de TUNEL o índice apoptótico. Os dados foram comparados por análise de variância e as médias por teste de Student Newman Keuls. A expressão gênica foi avaliada pelo software REST®. Os valores são mostrados como média±erro padrão. Embriões com 8 ou mais células produziram maiores (P<0,01) taxas de blastocistos no D7 e D8 do que os embriões com menos de oitos células mostrando seu maior potencial de desenvolvimento, independentemente se foram partenogenéticos ou fecundados. Embriões do grupo Part&#8805;8 apresentaram maior (P<0,05) taxa de blastocistos no D7 (63,6±3,4%) que os FIV&#8805;8 (45,3±8,9%), porém a taxa no D8 foi semelhante (56,7±3,0% e 44,2±8,9% para Part&#8805;8 e FIV&#8805;8, respectivamente; (P<0,05). Não houve diferença (P<0,05) quanto às taxas de blastocistos no D7 e D8 entre embriões com menos de 8 células oriundos da partenogênese ou fecundação. Não houve diferença quanto ao número total de células (92,0±3,4; 102,30±4,8), células apoptóticas (10,8±1,22; 9,5±1,07) e índice apoptótico (11,4±1,27; 9,8±1,8) nos blastocistos partenogenéticos e FIV, respectivamente. Nos embriões com 8 células analisados, houve subexpressão dos genes DNAJB1, HSPA1L, HSF1, GLUT1 nos Part em relação aos FIV, enquanto o HSF2 esteve sobrexpresso nos Part quando comparado aos FIV. Conclui-se que embriões partenogenéticos bovinos possuem diferenças se comparados aos embriões fecundados in vitro quanto à capacidade de desenvolvimento pré-implantacional in vitro e expressão gênica, o que pode limitar o uso da partenogênese em estudos sobre desenvolvimento embrionário.

Made available in DSpace on 2016-05-02T13:55:48Z (GMT). No. of bitstreams: 1 Tochimara Aparecida Myauchi Dissertacao.pdf: 1469858 bytes, checksum: d9554237da81a72762ef7629c0dfa1b8 (MD5) Previous issue date: 2012-09-28 / Coordenacao de Aperfeicoamento de Pessoal de Nïvel Superior / Parthenogenesis has been described as an alternative method to produce embryos for studies with embryonic cells, particularly in humans which have a restriction of fertilized embryos. Procedures used in parthenogenesis are also required to produce embryos by somatic cell nuclear transfer in domestic species. However, many biological, cellular and molecular aspects of parthenogenetic embryos are still unknown. This study aimed to compare the kinetics of development, apoptosis rate and gene expression of stress and celular metabolism in bovine parthenogenetic embryos and embryos fertilized in vitro. Oocytes (n = 1541) obtained from slaughterhouse ovaries were matured in vitro and submitted to parthenogenetic activation (4.62 µM ionomycin for 5 min followed by 2 mM 6-DMAP for 4 h) or in vitro fertilization (2 x 106 sperm/mL for 20h with semen from a single departure). 72h postactivation/fertilization (hpaf), embryos (8-cell) were frozen for subsequent analysis of gene expression and another part has been separated into groups of high or low potential of development: Part &#8805; 8 - parthenogenetic embryos with 8 or more cells (high development potential); Part < 8 - embryos less than 8 cells (low development potential); &#8805; 8 IVF - in vitro fertilized embryos with 8 or more cells; and IVF < 8 - embryos with less than 8 cells. Embryos were cultured in CR2aa medium with 2.5% BFS in 5% CO2, 5% O2, 90% N2 at 38.5° C and were evaluated rates of blastocyst 168hpaf (D7) and 196hpaf (D8). 8-cell embryos obtained after 72hpfa were analyzed for gene expression. D8 blastocysts were fixed and subsequently apoptotic index was analyzed by TUNEL. Data were compared by analysis of variance and means by Student Newman Keuls test. Gene expression was evaluated by REST® software. Values are shown as mean ± standard error. Embryos with 8 or more cells produced higher (P < 0.01) blastocysts rates in D7 and D8 compared to embryos with less than eight cells, showing its greatest development potential, regardless if they were parthenogenetic or fertilized. Embryos of Part &#8805; 8 group had higher (P <0.05) blastocyst rate in D7 (63.6 ± 3.4%) than IVF &#8805; 8 (45.3 ± 8.9%), but at D8 rate was similar (56.7 ± 3.0% and 44.2 ± 8.9% for Part &#8805; 8 and IVF &#8805; 8, respectively; P <0.05). There was no difference (P < 0.05) on blastocyst rates at D7 and D8 between embryos with less than 8 cells derived from parthenogenesis or fertilization. There was no difference in total cell number (92.0 ± 3.4, 102.30 ± 4.8), apoptotic cells (10.8 ± 1.22, 9.5 ± 1.07) and apoptotic index (11.4 ± 1.27, 9.8 ± 1.8) for parthenogenetic and IVF blastocysts, respectively. In 8-cell embryos analyzed, there was a subexpression of genes DNAJB1, HSPA1L, HSF1 and GLUT1 for Part in relation to FIV, while HSF2 was overexpressed in Part compared to FIV. In conclusion, bovine parthenogenetic embryos differ to embryos fertilized in vitro in hability of preimplantation development in vitro and gene expression, which may limit the use of parthenogenesis in studies of embryonic development. / A partenogênese tem sido descrita como um método alternativo para produzir embriões para estudos com células embrionárias, principalmente em humanos para os quais existe a restrição do uso de embriões fecundados. Procedimentos utilizados na partenogênese são também necessários para se produzir embriões por transferência nuclear com células somáticas nas espécies domésticas. Contudo, muitos aspectos biológicos, celulares e moleculares dos embriões partenogenéticos ainda são desconhecidos. Este estudo objetivou comparar a cinética do desenvolvimento, índice de apoptose e expressão de genes de estresse e metabolismo celular em embriões bovinos partenogenéticos e embriões fecundados in vitro. Oócitos (n=1541) obtidos de ovários de matadouro foram maturados in vitro e submetidos à ativação partenogenética (4,62 µM ionomicina por 5 min seguido de 2 mM 6-DMAP por 4h) ou fecundação in vitro (2 x 106 espermatozoides/ml por 20h, com sêmen de uma única partida). Com 72h pós-ativação/fecundação (hpaf) parte dos embriões (8 células) foram congelados para posterior análise da expressão gênica e outra parte foi separada em grupos de alto ou baixo potencial de desenvolvimento: Part&#8805;8 - embriões partenogenéticos com 8 ou mais células (alto potencial de desenvolvimento); Part<8 - embriões com menos de 8 células (baixo potencial de desenvolvimento); FIV&#8805;8 - embriões fecundados in vitro com 8 ou mais células; e FIV<8: embriões com menos de 8 células. Os embriões foram cultivados em meio CR2aa com 2,5% SFB em 5%CO2, 5%O2, 90% N2 a 38,5ºC e avaliadas as taxas de blastocistos com 168hpaf (D7) e 196hpaf (D8). Embriões com 8 células obtidos após 72hpfa foram analisados quanto à expressão gênica. Blastocistos em D8 foram fixados e posteriormente foram avaliados pela técnica de TUNEL o índice apoptótico. Os dados foram comparados por análise de variância e as médias por teste de Student Newman Keuls. A expressão gênica foi avaliada pelo software REST®. Os valores são mostrados como média±erro padrão. Embriões com 8 ou mais células produziram maiores (P<0,01) taxas de blastocistos no D7 e D8 do que os embriões com menos de oitos células mostrando seu maior potencial de desenvolvimento, independentemente se foram partenogenéticos ou fecundados. Embriões do grupo Part&#8805;8 apresentaram maior (P<0,05) taxa de blastocistos no D7 (63,6±3,4%) que os FIV&#8805;8 (45,3±8,9%), porém a taxa no D8 foi semelhante (56,7±3,0% e 44,2±8,9% para Part&#8805;8 e FIV&#8805;8, respectivamente; (P<0,05). Não houve diferença (P<0,05) quanto às taxas de blastocistos no D7 e D8 entre embriões com menos de 8 células oriundos da partenogênese ou fecundação. Não houve diferença quanto ao número total de células (92,0±3,4; 102,30±4,8), células apoptóticas (10,8±1,22; 9,5±1,07) e índice apoptótico (11,4±1,27; 9,8±1,8) nos blastocistos partenogenéticos e FIV, respectivamente. Nos embriões com 8 células analisados, houve subexpressão dos genes DNAJB1, HSPA1L, HSF1, GLUT1 nos Part em relação aos FIV, enquanto o HSF2 esteve sobrexpresso nos Part quando comparado aos FIV. Conclui-se que embriões partenogenéticos bovinos possuem diferenças se comparados aos embriões fecundados in vitro quanto à capacidade de desenvolvimento pré-implantacional in vitro e expressão gênica, o que pode limitar o uso da partenogênese em estudos sobre desenvolvimento embrionário.

Made available in DSpace on 2016-05-02T13:55:51Z (GMT). No. of bitstreams: 1 TochimaraAparecidaMiyauchi-dissertacao.pdf: 849651 bytes, checksum: ef29d9bc544c7cef9bb8b27b98b198ea (MD5) Previous issue date: 2012-09-28 / Coordenacao de Aperfeicoamento de Pessoal de Nïvel Superior / Parthenogenesis has been described as an alternative method to produce embryos for studies with embryonic cells, particularly in humans which have a restriction of fertilized embryos. Procedures used in parthenogenesis are also required to produce embryos by somatic cell nuclear transfer in domestic species. However, many biological, cellular and molecular aspects of parthenogenetic embryos are still unknown. This study aimed to compare the kinetics of development, apoptosis rate and gene expression of stress and celular metabolism in bovine parthenogenetic embryos and embryos fertilized in vitro. Oocytes (n = 1541) obtained from slaughterhouse ovaries were matured in vitro and submitted to parthenogenetic activation (4.62 µM ionomycin for 5 min followed by 2 mM 6-DMAP for 4 h) or in vitro fertilization (2 x 106 sperm/mL for 20h with semen from a single departure). 72h postactivation/fertilization (hpaf), embryos (8-cell) were frozen for subsequent analysis of gene expression and another part has been separated into groups of high or low potential of development: Part &#8805; 8 - parthenogenetic embryos with 8 or more cells (high development potential); Part < 8 - embryos less than 8 cells (low development potential); &#8805; 8 IVF - in vitro fertilized embryos with 8 or more cells; and IVF < 8 - embryos with less than 8 cells. Embryos were cultured in CR2aa medium with 2.5% BFS in 5% CO2, 5% O2, 90% N2 at 38.5° C and were evaluated rates of blastocyst 168hpaf (D7) and 196hpaf (D8). 8-cell embryos obtained after 72hpfa were analyzed for gene expression. D8 blastocysts were fixed and subsequently apoptotic index was analyzed by TUNEL. Data were compared by analysis of variance and means by Student Newman Keuls test. Gene expression was evaluated by REST® software. Values are shown as mean ± standard error. Embryos with 8 or more cells produced higher (P < 0.01) blastocysts rates in D7 and D8 compared to embryos with less than eight cells, showing its greatest development potential, regardless if they were parthenogenetic or fertilized. Embryos of Part &#8805; 8 group had higher (P <0.05) blastocyst rate in D7 (63.6 ± 3.4%) than IVF &#8805; 8 (45.3 ± 8.9%), but at D8 rate was similar (56.7 ± 3.0% and 44.2 ± 8.9% for Part &#8805; 8 and IVF &#8805; 8, respectively; P <0.05). There was no difference (P < 0.05) on blastocyst rates at D7 and D8 between embryos with less than 8 cells derived from parthenogenesis or fertilization. There was no difference in total cell number (92.0 ± 3.4, 102.30 ± 4.8), apoptotic cells (10.8 ± 1.22, 9.5 ± 1.07) and apoptotic index (11.4 ± 1.27, 9.8 ± 1.8) for parthenogenetic and IVF blastocysts, respectively. In 8-cell embryos analyzed, there was a subexpression of genes DNAJB1, HSPA1L, HSF1 and GLUT1 for Part in relation to FIV, while HSF2 was overexpressed in Part compared to FIV. In conclusion, bovine parthenogenetic embryos differ to embryos fertilized in vitro in hability of preimplantation development in vitro and gene expression, which may limit the use of parthenogenesis in studies of embryonic development. / A partenogênese tem sido descrita como um método alternativo para produzir embriões para estudos com células embrionárias, principalmente em humanos para os quais existe a restrição do uso de embriões fecundados. Procedimentos utilizados na partenogênese são também necessários para se produzir embriões por transferência nuclear com células somáticas nas espécies domésticas. Contudo, muitos aspectos biológicos, celulares e moleculares dos embriões partenogenéticos ainda são desconhecidos. Este estudo objetivou comparar a cinética do desenvolvimento, índice de apoptose e expressão de genes de estresse e metabolismo celular em embriões bovinos partenogenéticos e embriões fecundados in vitro. Oócitos (n=1541) obtidos de ovários de matadouro foram maturados in vitro e submetidos à ativação partenogenética (4,62 µM ionomicina por 5 min seguido de 2 mM 6-DMAP por 4h) ou fecundação in vitro (2 x 106 espermatozoides/ml por 20h, com sêmen de uma única partida). Com 72h pós-ativação/fecundação (hpaf) parte dos embriões (8 células) foram congelados para posterior análise da expressão gênica e outra parte foi separada em grupos de alto ou baixo potencial de desenvolvimento: Part&#8805;8 - embriões partenogenéticos com 8 ou mais células (alto potencial de desenvolvimento); Part<8 - embriões com menos de 8 células (baixo potencial de desenvolvimento); FIV&#8805;8 - embriões fecundados in vitro com 8 ou mais células; e FIV<8: embriões com menos de 8 células. Os embriões foram cultivados em meio CR2aa com 2,5% SFB em 5%CO2, 5%O2, 90% N2 a 38,5ºC e avaliadas as taxas de blastocistos com 168hpaf (D7) e 196hpaf (D8). Embriões com 8 células obtidos após 72hpfa foram analisados quanto à expressão gênica. Blastocistos em D8 foram fixados e posteriormente foram avaliados pela técnica de TUNEL o índice apoptótico. Os dados foram comparados por análise de variância e as médias por teste de Student Newman Keuls. A expressão gênica foi avaliada pelo software REST®. Os valores são mostrados como média±erro padrão. Embriões com 8 ou mais células produziram maiores (P<0,01) taxas de blastocistos no D7 e D8 do que os embriões com menos de oitos células mostrando seu maior potencial de desenvolvimento, independentemente se foram partenogenéticos ou fecundados. Embriões do grupo Part&#8805;8 apresentaram maior (P<0,05) taxa de blastocistos no D7 (63,6±3,4%) que os FIV&#8805;8 (45,3±8,9%), porém a taxa no D8 foi semelhante (56,7±3,0% e 44,2±8,9% para Part&#8805;8 e FIV&#8805;8, respectivamente; (P<0,05). Não houve diferença (P<0,05) quanto às taxas de blastocistos no D7 e D8 entre embriões com menos de 8 células oriundos da partenogênese ou fecundação. Não houve diferença quanto ao número total de células (92,0±3,4; 102,30±4,8), células apoptóticas (10,8±1,22; 9,5±1,07) e índice apoptótico (11,4±1,27; 9,8±1,8) nos blastocistos partenogenéticos e FIV, respectivamente. Nos embriões com 8 células analisados, houve subexpressão dos genes DNAJB1, HSPA1L, HSF1, GLUT1 nos Part em relação aos FIV, enquanto o HSF2 esteve sobrexpresso nos Part quando comparado aos FIV. Conclui-se que embriões partenogenéticos bovinos possuem diferenças se comparados aos embriões fecundados in vitro quanto à capacidade de desenvolvimento pré-implantacional in vitro e expressão gênica, o que pode limitar o uso da partenogênese em estudos sobre desenvolvimento embrionário.

The development of mouse (Mus musculus) haploid parthenogenetic embryos does not reach the same level as normal embryos. The aim of this diploma thesis was to find out whether haploid parthenogenetic embryos of mice differ in the nucleocytoplasmic ratio. The volume of the nucleus increases with ploidity. The nucleocytoplasmic ratios of haploid embryos do not significantly change between the two-cell and four-cell stage (p = 0.052), there is a significant difference (p < 0.001) for diploid and tetraploid embryos. Non-standard nucleocytoplasmic ratio could be related to the problematic development. Understanding the regulation of preimplantational development of parthenogenetic embryos will increase the efficiency of haploid embryonic stem cell derivation.

Background. The phylum Tardigrada consists of over 1,300 species that inhabit terrestrial, freshwater and marine environments throughout the world. In terrestrial habitats they live primarily in mosses, lichens, leaf litter and soil, whereas tardigrades in freshwater and marine environments are mainly found in sediments and on aquatic plants. More than 65 species have been previously reported in the state of Tennessee, USA. Methods. Tardigrades present in moss cushions (Grimmia sp.) collected from a xerothermic habitat on the East Tennessee State University campus, Johnson City, TN, USA, were extracted, mounted on slides, identified, and counted. Additional samples of fresh dried moss were used for integrative analyses, including morphological analysis with phase contrast (PCM) and scanning electron microscopy (SEM), as well as molecular analyses of COI, 18S rRNA, 28S rRNA, and ITS-2 of the Macrobiotus and Milnesium species. Results. Five species were found, including two species new to science: Viridiscus miraviridis sp. nov. and Macrobiotus basiatus sp. nov. Viridiscus miraviridis sp. nov. differs from other members of the genus mainly by having a different type of dorsal cuticle and some other, more subtle, morphometric characters. In addition to the two new species, Viridiscus perviridis and Viridiscus viridissimus were present, and males of Vir. viridissimus were found for the first time, the first record of males in the genus Viridiscus. Macrobiotus basiatus sp. nov. is most similar to Macrobiotus nelsonae, but it differs from Mac. nelsonae mainly by the stylet supports being situated in a more anterior position, shorter and narrower egg processes, and a smaller number of areoles around the egg processes. Moreover, the identification of Milnesium inceptum was confirmed as the first record for the USA by analysis of COI.

Multiple stressors have caused a decline in coral populations. Broadcast spawning corals once dominated the Florida Reef Tract (FRT), but since their decline, smaller brooding corals, soft corals, and macroalgae are replacing them. Brooding corals are more resilient to current threats in part because they are reproductive throughout much of the year and their larvae are competent to settle after release. Despite the ubiquity of brooders on Florida reefs, much of their reproductive strategy remains unknown. This study aimed to examine paternity as a function of colony size and density in Porites astreoides, a common brooding coral in the FRT. Porites astreoides colonies were configured in arrays at three densities that were replicated three times. A focal colony was surrounded by six other colonies, separated from the focal colony at different distances (1m, 7m, and 15m) representing high, moderate, and low population densities, respectively. All arrays were placed in the field but were separated from the reef and naturally occurring P. astreoides colonies by at least 50 m. Four days before the new moon, colonies were transported to the laboratory for larval collection. Over a four day period, a total of 3,184 larvae were collected from 24 colonies, 13 of which released larvae over consecutive days. The resulting larvae were genotyped using seven microsatellite markers. All larvae had the exact genotypes of the colony from which the larvae were collected, i.e. maternal- egg donor. This suggested the larvae were parthenogenically produced and no sperm was used to fertilize the eggs. This is the first study to suggest that parthenogenesis is occurring in P. astreoides. In today's oceans that have been depleted of corals, parthenogenesis may be an advantageous reproductive strategy used to boost populations. However, parthenogenesis reduces the genetic diversity which could hinder successful sexual reproduction in the future causing fragmented populations.

In the modern poultry industry, intense genetic selection for meat production has negatively influenced the reproductive performance of commercial birds. Parthenogenesis, embryonic development in unfertilized eggs without any sperm-egg interactions, is known to hinder the normal fertilization process and could be one of the reasons for this reduced reproductive performance in the poultry industry. Therefore, the overall objective of this research was to gain a better understanding of the process of parthenogenesis using Chinese painted quail as the model. Studies on Chinese painted quail reproduction revealed that they are very inefficient in sustained sperm storage and that number of sperm penetrating the egg and subsequent embryonic development potentially alter egg transit time through the oviduct. This poor sperm storage capacity and high sperm-egg interaction requirement might be responsible for the occurrence of parthenogenesis in this species; and in fact, this makes Chinese painted quail an excellent choice for parthenogenesis research. Further, dams selected for parthenogenesis as well as embryonic development, including parthenogen size, alter egg components by possibly delaying the transit time of the egg through the oviduct. Also, both dams and sires selected for the parthenogenesis trait appear to influence their progenies performance, including 1st wk mortality and occurrence of parthenogenesis. Additionally, vaccination of virgin hens with live pigeon pox virus increases parthenogenesis as well as parthenogen size and livability by the direct action of the virus on the embryo. Moreover, live Newcastle disease virus under in vitro conditions was found to have similar effects on the embryo. Because parthenogenesis exists in the modern poultry industry, even the accidental selection of the trait in either males or females could have a negative impact on overall chick production and performance. Also, as vaccination is a routine practice in the industry, it is possible that vaccination of birds that carry the trait will reduce fertility and hatchability due to enhanced parthenogenesis. Overall, currently it appears that, parthenogenesis is adversely affecting the poultry industry; and therefore, additional research on the accurate determination of losses in the poultry industry due to parthenogenesis could further benefit the industry.

Three aspects of Xanthoparmelia cumberlandia and Xanthoparmelia coloradoënsis populations found at two elevations are explored: clustering of secondary chemicals and the resulting implications for taxonomic distinctions, the usefulness of thallus size as an indirect measure of sexual fecundity, and the frequency of sexual reproduction. First, we use clustering of 46 chemicals produced by X. cumberlandia and X. coloradoënsis to evaluate the adequacy of the current taxonomic distinction between them. Using principal components analysis and UPGMA, we find that the currently recognized species boundaries indicated by the presence of stictic acid in X. cumberlandia and salazinic acid in X. coloradoënsis are supported by distinct differences in their chemotypes (combinations of secondary chemicals). Norstictic acid, which the literature also associates with X. cumberlandia, is found frequently in both X. cumberlandia and X. coloradoënsis, and is not a good distinguishing characteristic. No chemical difference between sexually fecund and sterile individuals was found. Second, we test the claim that thallus size can be used as an indirect measure of sexual fecundity. By comparing the number of apothecia, the total area of the apothecia, and the presence or absence of apothecia with thallus area, we found positive correlations between these measures of sexual fecundity and thallus size which are statistically significant. However, the total variation explained by these predictors is limited, and is significantly affected by elevation and micro-environmental features such as proximity to trees. We conclude that size is not a reliable synonym for sexual fecundity in X. cumberlandia and X. coloradoënsis. Third, we make inferences concerning the frequency of sexual reproduction based on the frequency of sexual structures, rare chemicals, and unique chemotypes. We predicted that sexual reproduction would be more frequent at lower elevations, consistent with a common pattern found in plants and animals. The frequency of sexual structures indicates that sexual reproduction is more common at the lower elevation, while frequency of rare chemicals and chemotypes implies that outcrossing is more common at the upper elevation. Since these indicators lead to opposing conclusions, we encourage the use of molecular markers to estimate the frequency of outcrossing directly.

Previous work has shown that active noise control (ANC) can be applied to axial cooling fans. Optimization of the control source and error sensor placement is desired to maximize the attenuation using ANC. A genetic algorithm was developed to find the optimal placement of control sources for a given primary source. The optimal configuration of control sources around a single primary source was shown to be a linear arrangement of the sources. This holds true for both two-dimensional as well as three-dimensional configurations. The higher-order radiation of the linear arrangement has also been verified experimentally, but the improvement in the experimental apparatus was not as dramatic as the theoretical model. Multiple flow visualization techniques have been used to find optimal near field error sensor locations. When there is little obstruction to the flow field of the fan, minimal airflow is found along the near field null that is created by minimizing the sound power of the system. Surface mounting of the error sensors can lead to a small increase in the signal-to-noise ratio of the error sensors if vortices exist in the near field of the fan due to obstructions in the main flow. It has also been shown that the introduction of the ANC system does not affect the flow field of the fan.

The Tardigrada are hydrophilous, segmented, molting micrometazoans that occupy a diversity of niches in freshwater, marine, and terrestrial habitats. A sister group of the arthropods, this phylum of bilaterally symmetrical lobopods, most less than 1 mm in length, have a hemocoel, a complete digestive tract, a dorsal gonad with one or two gonoducts, and a dorsal lobed brain with a ventral nerve cord and five ganglia. About 1000 species have been described based on the morphology of sclerified structures, especially the claws and buccal-pharyngeal apparatus. Reproduction occurs through fertilized or unfertilized eggs, with individuals being either gonochoric, unisexual, or hermaphroditic, and eggs are deposited either freely or within the shed exuvium. Parthenogenesis, very frequent in limnic and terrestrial tardigrades, allows them to colonize new territories by passive dispersal of a single individual. Quiescence (cryptobiosis: anhydrobiosis, anoxybiosis, cryobiosis, and osmobiosis) and diapause (encystment and resting eggs) occur during the tardigrade life history. Ecological parameters and global distribution patterns are poorly known or understood. Methods for collection, microscopy, and culturing have been developed.