Altas taxas de mortalidade embrionária ocorrem no período de implantação em embriões provenientes de IATF e FIV. Acredita-se que a falha no desenvolvimento do saco vitelino possa estar relacionada com estas perdas embrionárias. O objetivo deste trabalho foi descrever as características macroscópicas, microscópicas e expressão gênica do processo de involução do saco vitelino, visto que os dados da literatura são contraditórios ao descrever os processos de formação e involução do saco vitelino em embriões provenientes de IATF e FIV. Foram utilizados 19 embriões de IATF e 16 de FIV, provenientes da Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brasil. Observou-se que não houve diferença significativa no tamanho dos embriões nas idades de 25 (p=0,1), 35 (p=0,1), 40 (p=0,4) e 45 (p=0,36) dias, quando comparados os grupos de IATF e FIV. Entretanto entre os embriões de IATF e FIV com 30 dias de gestação, apresentaram diferença estatística (p= 0,03). A curva de crescimento (Crow-Rump) aumentou gradativamente e linearmente em função da idade gestacional. Macroscopicamente, observou-se que aos 25 dias de gestação, o saco vitelino apresentava formato de T e coloração translúcida. Uma porção central se encontrava próxima a região ventral do embrião emitindo duas extremidades que eram finas membranas, estas acompanhavam o alantóide juntamente com os vasos umbilicais. Aos 30 a 35 dias, apresentava coloração amarelada, sua parte central e extremidades bem diminuídas mostrando sinais de um início de regressão. Aos 40 a 45 dias, o seu processo de involução estava caracterizado, com extremidades enoveladas e região central assemelhando-se a um pequeno grão achatado que se encontrava em contato com o âmnio. Microscopicamente, o saco vitelino era formado por três camadas: o endoderma, camada única que reveste a cavidade vitelínica, uma camada intermediária mesenquimal vascular e o mesotélio, camada simples voltada ao exoceloma. O epitélio do saco vitelino formava dobras ou criptas que se projetavam para a luz. Imunohistoquímica notou-se marcação para os anticorpos VEGF, Flt-1 e KDR. Observou-se que não houve diferença significativa na expressão do mRNA do VEGF, KDR e Flt-1 entre os grupos e nas idades gestacionais analisadas, aos 30 dias notou-se uma tendência entre os grupos em relação a expressão do Flt-1. Os fatores de crescimento b FGF, FGFR-1 e FGFR-2, não apresentaram diferenças em suas expressões, somente aos 35 dias de gestação para o mRNA FGFR-1. Conclui-se que o saco vitelino está intensamente relacionado com o desenvolvimento embrionário, na qual sua involução e desaparecimento relacionam-se com o crescimento e \"maturação\" embrionária. / High mortality rates occur during embryonic implantation embryos from IVF and FTAI. It is believed that the failure in the development of the yolk sac may be related to these embryonic losses. The objective of this study was to describe the macroscopic, microscopic and mRNA expression features of the yolk sac involution process, since the literature relates contradictory information regarding the processes of formation and involution of the yolk sac in IVF and TAI embryos. Nineteen embryos were used for FTAI and 16 IVF; the specimes were obtained from the Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brazil. No significant differences were observed in the size of embryos aged 25 (p = 0.1), 35 (p = 0.1), 40 (p = 0.4) and 45 (p = 0.36) days in the groups of IVF and FTAI. However between FTAI and IVF embryos with 30 days of gestation, a difference (p = 0.03) was detectable. The growth curve (Crow-Rump) increased gradually and linearly with gestational age. Macroscopically, it was observed that after 25 days of gestation, the yolk sac was \"T-shaped\" and transparent colored. A central portion was observed in the vicinity of the ventral ends of the embryo exhibiting two thin membranes, which accompanied the allantois with the umbilical vessels. At 30 to 35 days this portion becomes yellowish and its central part together with the extremities was reduced displaying clearly signs of regression. At 40 to 45 days, the process of involution was presented in a curled fashion and ends with the central region resembling a flat small grain in contact with the amnion. Microscopically, the yolk sac was formed by three layers: the endoderm, single layer lining the vitelline cavity, an intermediate layer and vascular mesenchymal mesothelium, exoceloma focused on single layer. The epithelium of the yolk sac formed folds or crypts, which projected to the light. Immunohistochemistry marking was noted for antibodies VEGF, Flt-1 and KDR. No significant difference were observed in mRNA expression of VEGF, KDR and Flt-1 between groups and the gestational ages analyzed at 30 days there has been a trend among the groups regarding the expression of Flt-1. The growth factors bFGF, FGFR-1 and FGFR-2 showed no differences in their expressions, with the exception of 35 days of gestation for the m-RNA FGFR 1. It was concluded that the yolk sac is strongly related to embryonic development, in which the involution and disappearance are related to growth and \"maturity\" stage.

A inseminação artificial em tempo fixo (IATF), juntamente com a inseminação artificial pós-cervical (IAPC), permite uma redução expressiva da mão de obra e uma melhor utilização dos machos geneticamente superiores. O objetivo deste estudo foi avaliar a eficiência do eCG associado agonista de GnRH (buserelina) ou apenas buserelina para indução e sincronização da ovulação em porcas desmamadas e submetidos a apenas uma IATF. Um total de 495 femeas suínas desmamadas foram utilizadas, cuja detecção de estro foi realizada uma vez ao dia (07:30). As fêmeas foram divididas em três tratamentos: Controle (n = 165) - a primeira IA foi realizada no início do estro (0 h) e repetida a cada 24 h, posteriormente, durante o estro; Tratamento 2: eCG + GnRH (n = 165) – femeas receberam uma injeção intramuscular de 600 UI de eCG após o desmame, e uma injeção intramuscular de buserelina (10 μg) 86-89 h após a administração de eCG; Tratamento 3: GnRH - fêmeas receberam uma injeção intramuscular de buserelina (10 μg) 86-89 h após o desmame. Nos três grupos era realizada a exposição ao macho no dia seguinte ao desmame uma vez ao dia, e aquelas fêmeas que demostrassem estro eram retiradas dos protocolos de IATF. Os grupos tratados receberam uma única IA, 118-120 h após o desmame (30-33 h após a buserelina). A IAPC contou com doses homospérmicas (1,5 x 109 células de espermáticas/50 ml) que foram utilizadas nas femeas em todos os três tratamentos. Não houve diferenças entre os tratamentos quanto ao número de femeas em estro até o terceiro dia (P>0,05). O intervalo entre o desmame e a ovulação foi significativamente maior (P< 0,05) no grupo controle (141,5 ± 1,58 h) comparativamente aos tratamentos eCG+GnRH (133,3 ± 1,60 h) e GnRH (135,9 ± 1,57 h). As femeas do tratamento eCG+GnRH ovularam em média antes (P<0,05) em relação ao grupo GnRH (44,5 ± 1,74 vs 48,2 ± 1,73 h). A taxa de parto foi superior (P<0,05) no grupo de controle, comparativamente aos tratamentos eCG + GnRH e GnRH, mas não houve diferença (P>0,05) em leitões nascidos totais entre os tratamentos. O uso do GnRH, com ou sem a administração prévia de eCG, antecipa a ovulação nas fêmeas suínas desmamadas. Nas fêmeas tratadas com eCG+GnRH ou GnRH, há em maior percentual de fêmeas inseminadas fora do momento ideal, em relação à ovulação, o que pode prejudicar o desempenho reprodutivo, sendo necessários ajustes no momento de aplicação dos hormônios ou no momento da inseminação para estudos subsequentes. / Fixed-time artificial insemination (FTAI) together with post-cervical artificial insemination (PCAI), allows an expressive reduction in labor requirements and a wider use of higher indexing boars. The aim of this study was to evaluate the efficiency of eCG agonist of GnRH (buserelin) or just buserelin for induction and synchronization of ovulation in weaned sows and submitted to FTAI .A total of 495 weaned sows whose estrus detection was performed once daily (07:30 A.M), starting a day after weaning. The sows were allocated into three treatment groups: Control (n=165) – the first AI was performed at estrus onset (0 h) and repeated every 24 h thereafter during estrus; Treatment 2: eCG + GnRH (n=165) – sows received an intramuscular injection of 600 UI eCG after weaning, and an intramuscular injection of buserelin (10 μg) 86-89 h after eCG administration; Treatment 3: GnRH - sows received an intramuscular injection of buserelina (10 μg) 86-89 h after weaning. . In the three groups it was performed boar exposure in the following day to weaning once a day, and those sows that demonstrated oestrus were removed from FTAI protocols. Treated groups received a single AI, 118-120 h after weaning (30-33 h after buserelin). PCAI with homospermic doses (1.5 x 109 total of sperm cells/50 ml) were respectively performed in sows in all treatment group. There were no differences (P> 0,05) in number of sows that showed oestrus until the third day between treatments. The interval between weaning and ovulation was significantly higher (P< 0,05) in the control group (141,5 ± 1,58 h) comparatively the treatments eCG+GnRH (133,3 ± 1,60 h) and GnRH (135,9 ± 1,57 h). Sows of treatment eCG+GnRH ovulated earlier (P< 0,05) comparing to the GnRH group (44,5 ± 1,74 vs 48,2 ± 1,73 h). Higher (P<0,05) farrowing rate in control group comparatively to treatments eCG+GnRH e GnRH was observed, but there were no differences (P> 0,05) in total piglets born between treatments. Use of GnRH, with or without previous administration of eCG anticipates ovulation in sows weaned, in the sows treated with eCG + GnRH or GnRH, there is a greater percentage of females inseminated outside the ideal time, with respect to ovulation, it can harm the reproductive performance, with necessary adjustments at the time of application hormones or at the time of insemination for subsequent studies.

Made available in DSpace on 2015-03-26T13:46:37Z (GMT). No. of bitstreams: 1 texto completo.pdf: 241274 bytes, checksum: 50b26ed9f1cc9fd1c0b7833ac2ec66ba (MD5) Previous issue date: 2006-07-31 / The effect of the hormonal induction was studied with 90 goats of the Toggenburg breed of three different categories (lactating, not lactating and nulliparous), in the following parameters: the average interval of the end of the hormonal treatment to the beginning of estrous (IE), the average duration of estrous (DE) and the gestation rate (GR). All the goats used in the study had been submitted to an pre-induction of estrous by artificial photoperiod per 60 days (14 of light and 10 of dark), and the hormonal inductions had occurred in three stages, with 65, 73 and 100 days after the end of the treatment with light. The hormonal induction consisted in the use of intravaginal sponges contends 60 mg of acetate of medroxiprogesterone (MAP, Day 0) per six days, in the intravulvo-submucosal administration of 37,5 &#956;g of synthetic prostaglandin, and in 200 UI of eCG (Day 4) intramuscular . In the animals of T1 and T2 the sponges were inserted and had been removed and hormones managed always in the morning and in the ones of T3 in the afternoon. The goats of T1 had been inseminated 11,3 ± 4,0 h after the beginning of estrous, and those that remained in estrous received a second insemination with 35,2 ± 6,0 h, those of T2 and T3 received two doses of semen each, witch these in fixed time with regard to the removal of the sponge, corresponding the 35,0 ± 2,2 h and 58,2 ± 2,1 h in T2, and 46,3 ± 1,4 h and 66,1 ± 2,3 h in T3. About to the interval estrous 1st insemination and estrous 2nd insemination in T2 was -10,7 ± 18,9 h and 12,5 ± 19,5 h, and T3 -8,3 ± 25,4 h and 14,4 ± 24.4 h. It had significant difference (P< 0,05) with relation to the intervals 1stAI, estrous 1stAI, 2ndAI and estrous 2ndAI in function of the treatments. The percentage of animals in estrous also differed (P< 0,05) between the treatments, T1 64,3 %, T2 96,1 % and T3 65,5 %. The IE was on 49,0 ± 22,0 h and DF 32,5 ± 19,6 h, not occurring difference between categories and treatments (P> 0,05), but yes with regard to the group of induction of estrous, being IE and DF 30,8 ± 12,9 h and 40,2 ± 25,0 in GI, 54,8 ± 13,9 h and 32,3 ± 13,3 h in GII, and 66,7 ± 23,2 h and 21,7 ± 13,3 h in GIII (P< 0,05). A negative correlation was detected (r-0,47) enters DF and IE (P< 0,05). The GR was of 21,0 % in T1, 30.8 % in T2 and 34.5 % in T3, not having difference (P> 0,05). (2) Interval from sponge removal to ovulation (IO), interval from estrous to ovulation (IEO) and interval from inseminations to ovulation. The transrectal ultrasonografic monitoring was made with a probe of 5 MHZ in 30 animals, witch 10 of each treatment of 1st group estrous induction, to each six hours from the sponge removal until the ovulation. The IO did not present difference (P> 0,05) between the treatments and categories, being 49,9 ± 8,2 h, 54,4 ± 10,1 h and 53,4 ± 12,3 h for T1, T2 and T3, respectively. The IEO also did not differ (P> 0,01), being 24,3 ± 6,7 h, 23,7 ± 12,3 h and 18,1 ± 26,3 h. through this monitoring we could observe that the inseminations had been made with - 12,1 ± 10,6 h and 11,4± 7,9 h in T1, -18,0 ± 10,6 h and 10,6 ± 12,4 h in T2, and - 6,4 ± 12,6 h and 10,6 ± 12,4 h in T3, not having difference (P> 0,05) in relation to the treatments. The ovulation rate was of 80 in T1, 100% in T2 and 100% in T3, revealing to be a good protocol for induction of estrous in goats out of the reproductive season. With these data about ovulation the TAI protocol could be adjusted, getting better results. / Estudou-se com 90 cabras da raça Toggenburg de três diferentes categorias (lactantes, não lactantes e nulíparas) o efeito da indução hormonal nos seguintes parâmetros: o intervalo médio do final do tratamento hormonal ao início do estro (IE), a duração média do estro (DE) e a taxa de gestação (TG). Todas as cabras utilizadas no estudo foram submetidas a uma pré-indução de estro pelo fotoperíodo artificial por 60 dias (14 de luz e 10 de escuro), e as induções hormonais ocorreram em três etapas, com 65, 73 e 100 dias após o termino do tratamento com luz. A indução hormonal consistiu no uso de esponjas intravaginais contendo 60 mg de acetato de medroxiprogesterona (MAP, Dia 0) por seis dias, na administração intravulvo-submucosal de 37,5 &#956;g de prostaglandina sintética, e intramuscular de 200 UI de eCG (Dia 4). Nos animais de T1 e T2 as esponjas foram inseridas e removidas e os hormônios administrados sempre pela manha, e nos de T3 pela tarde. As cabras pertencentes ao T1 foram inseminadas 11,3 ± 4,0 h após o início do estro, e aquelas que permaneceram em estro receberam uma segunda inseminação com 35,2 ± 6,0 h, as do T2 e T3 receberam duas doses de sêmen cada, sendo estas em tempo fixo com relação à remoção da esponja, correspondendo a 35,0 ± 2,2 h e 58,2 ± 2,1 h em T2, e 46,3 ± 1,4 h e 66,1 ± 2,3 h em T3. Quanto ao intervalo estro 1ª inseminação e estro 2ª inseminação em T2 foi de 10,7 ± 18,9 h e 12,5 ± 19,5 h, e T3 8,3 ± 25,4 h e 14,4 ± 24,4 h. Houve diferença significativa (P< 0,05) com relação aos intervalos 1ªIA, estro 1ªIA, 2ªIA e estro 2ªIA em função dos tratamentos. A percentagem de animais em estro também diferiu (P< 0,05) entre os tratamentos sendo, T1 64,3%, T2 96,1% e T3 65,5%. O IE foi em média 49,0 ± 22,0 h e a DE 32,5 ± 19,6 h, não ocorrendo diferença entre categorias e tratamentos (P> 0,05), mas sim com relação ao grupo de indução de estro, sendo IE e DE de 30,8 ± 12,9 h e 40,2 ± 25,0 em GI, 54, 8 ± 13,9 h e 32,3 ± 13,3 h em GII, e 66,7 ± 23,2 h e 21,7 ± 13,3 h em GIII (P< 0,05). Detectou-se uma correlação negativa (r-0,47) entre DE e IE (P< 0,05). A TG foi de 21,0% em T1, 30,8% em T2 e 34,5% em T3, não havendo diferença (P> 0,05). Intervalo da retirada da esponja à ovulação (IO), intervalo do estro a ovulação (IEO) e intervalo das inseminações à ovulação. Foi feito o monitoramento ultrassonográfico transretal com auxilio de uma probe de 5 MHZ de 30 animais, sendo 10 de cada tratamento do 1º grupo de indução de estro, a cada oito horas a partir da retirada da esponja ate a ovulação. O IO não apresentou diferença (P> 0,05) entre os tratamentos e categorias, sendo 49,9 ± 8,2 h, 54,4 ± 10,1 h e 53,4 ± 12,3 h para T1, T2 e T3, respectivamente. O IEO também não diferiu (P> 0,01), sendo 24,3 ± 6,7 h, 23,7 ± 12,3 h e 18,1 ± 26,3 h. Através desse monitoramento pudemos observar que as inseminações foram feitas com 12,1 ± 10,6 h e 11,4± 7,9 h em T1, -18,0 ± 10,6 h e 10,6 ± 12,4 h em T2, e 6,4 ± 12,6 h e 10,6 ± 12,4 h em T3, não havendo diferença (P> 0,05) com relação aos tratamentos. A taxa de ovulação foi de 80% em T1, 100% em T2 e 100% em T3, mostrando-se ser um bom protocolo para indução de estro em cabras fora da estação reprodutiva. Com esses dados sobre ovulação podese ajustar o protocolo de IATF, obtendo-se melhores resultados.

A inseminação artificial em tempo fixo (IATF), juntamente com a inseminação artificial pós-cervical (IAPC), permite uma redução expressiva da mão de obra e uma melhor utilização dos machos geneticamente superiores. O objetivo deste estudo foi avaliar a eficiência do eCG associado agonista de GnRH (buserelina) ou apenas buserelina para indução e sincronização da ovulação em porcas desmamadas e submetidos a apenas uma IATF. Um total de 495 femeas suínas desmamadas foram utilizadas, cuja detecção de estro foi realizada uma vez ao dia (07:30). As fêmeas foram divididas em três tratamentos: Controle (n = 165) - a primeira IA foi realizada no início do estro (0 h) e repetida a cada 24 h, posteriormente, durante o estro; Tratamento 2: eCG + GnRH (n = 165) – femeas receberam uma injeção intramuscular de 600 UI de eCG após o desmame, e uma injeção intramuscular de buserelina (10 μg) 86-89 h após a administração de eCG; Tratamento 3: GnRH - fêmeas receberam uma injeção intramuscular de buserelina (10 μg) 86-89 h após o desmame. Nos três grupos era realizada a exposição ao macho no dia seguinte ao desmame uma vez ao dia, e aquelas fêmeas que demostrassem estro eram retiradas dos protocolos de IATF. Os grupos tratados receberam uma única IA, 118-120 h após o desmame (30-33 h após a buserelina). A IAPC contou com doses homospérmicas (1,5 x 109 células de espermáticas/50 ml) que foram utilizadas nas femeas em todos os três tratamentos. Não houve diferenças entre os tratamentos quanto ao número de femeas em estro até o terceiro dia (P>0,05). O intervalo entre o desmame e a ovulação foi significativamente maior (P< 0,05) no grupo controle (141,5 ± 1,58 h) comparativamente aos tratamentos eCG+GnRH (133,3 ± 1,60 h) e GnRH (135,9 ± 1,57 h). As femeas do tratamento eCG+GnRH ovularam em média antes (P<0,05) em relação ao grupo GnRH (44,5 ± 1,74 vs 48,2 ± 1,73 h). A taxa de parto foi superior (P<0,05) no grupo de controle, comparativamente aos tratamentos eCG + GnRH e GnRH, mas não houve diferença (P>0,05) em leitões nascidos totais entre os tratamentos. O uso do GnRH, com ou sem a administração prévia de eCG, antecipa a ovulação nas fêmeas suínas desmamadas. Nas fêmeas tratadas com eCG+GnRH ou GnRH, há em maior percentual de fêmeas inseminadas fora do momento ideal, em relação à ovulação, o que pode prejudicar o desempenho reprodutivo, sendo necessários ajustes no momento de aplicação dos hormônios ou no momento da inseminação para estudos subsequentes. / Fixed-time artificial insemination (FTAI) together with post-cervical artificial insemination (PCAI), allows an expressive reduction in labor requirements and a wider use of higher indexing boars. The aim of this study was to evaluate the efficiency of eCG agonist of GnRH (buserelin) or just buserelin for induction and synchronization of ovulation in weaned sows and submitted to FTAI .A total of 495 weaned sows whose estrus detection was performed once daily (07:30 A.M), starting a day after weaning. The sows were allocated into three treatment groups: Control (n=165) – the first AI was performed at estrus onset (0 h) and repeated every 24 h thereafter during estrus; Treatment 2: eCG + GnRH (n=165) – sows received an intramuscular injection of 600 UI eCG after weaning, and an intramuscular injection of buserelin (10 μg) 86-89 h after eCG administration; Treatment 3: GnRH - sows received an intramuscular injection of buserelina (10 μg) 86-89 h after weaning. . In the three groups it was performed boar exposure in the following day to weaning once a day, and those sows that demonstrated oestrus were removed from FTAI protocols. Treated groups received a single AI, 118-120 h after weaning (30-33 h after buserelin). PCAI with homospermic doses (1.5 x 109 total of sperm cells/50 ml) were respectively performed in sows in all treatment group. There were no differences (P> 0,05) in number of sows that showed oestrus until the third day between treatments. The interval between weaning and ovulation was significantly higher (P< 0,05) in the control group (141,5 ± 1,58 h) comparatively the treatments eCG+GnRH (133,3 ± 1,60 h) and GnRH (135,9 ± 1,57 h). Sows of treatment eCG+GnRH ovulated earlier (P< 0,05) comparing to the GnRH group (44,5 ± 1,74 vs 48,2 ± 1,73 h). Higher (P<0,05) farrowing rate in control group comparatively to treatments eCG+GnRH e GnRH was observed, but there were no differences (P> 0,05) in total piglets born between treatments. Use of GnRH, with or without previous administration of eCG anticipates ovulation in sows weaned, in the sows treated with eCG + GnRH or GnRH, there is a greater percentage of females inseminated outside the ideal time, with respect to ovulation, it can harm the reproductive performance, with necessary adjustments at the time of application hormones or at the time of insemination for subsequent studies.

Altas taxas de mortalidade embrionária ocorrem no período de implantação em embriões provenientes de IATF e FIV. Acredita-se que a falha no desenvolvimento do saco vitelino possa estar relacionada com estas perdas embrionárias. O objetivo deste trabalho foi descrever as características macroscópicas, microscópicas e expressão gênica do processo de involução do saco vitelino, visto que os dados da literatura são contraditórios ao descrever os processos de formação e involução do saco vitelino em embriões provenientes de IATF e FIV. Foram utilizados 19 embriões de IATF e 16 de FIV, provenientes da Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brasil. Observou-se que não houve diferença significativa no tamanho dos embriões nas idades de 25 (p=0,1), 35 (p=0,1), 40 (p=0,4) e 45 (p=0,36) dias, quando comparados os grupos de IATF e FIV. Entretanto entre os embriões de IATF e FIV com 30 dias de gestação, apresentaram diferença estatística (p= 0,03). A curva de crescimento (Crow-Rump) aumentou gradativamente e linearmente em função da idade gestacional. Macroscopicamente, observou-se que aos 25 dias de gestação, o saco vitelino apresentava formato de T e coloração translúcida. Uma porção central se encontrava próxima a região ventral do embrião emitindo duas extremidades que eram finas membranas, estas acompanhavam o alantóide juntamente com os vasos umbilicais. Aos 30 a 35 dias, apresentava coloração amarelada, sua parte central e extremidades bem diminuídas mostrando sinais de um início de regressão. Aos 40 a 45 dias, o seu processo de involução estava caracterizado, com extremidades enoveladas e região central assemelhando-se a um pequeno grão achatado que se encontrava em contato com o âmnio. Microscopicamente, o saco vitelino era formado por três camadas: o endoderma, camada única que reveste a cavidade vitelínica, uma camada intermediária mesenquimal vascular e o mesotélio, camada simples voltada ao exoceloma. O epitélio do saco vitelino formava dobras ou criptas que se projetavam para a luz. Imunohistoquímica notou-se marcação para os anticorpos VEGF, Flt-1 e KDR. Observou-se que não houve diferença significativa na expressão do mRNA do VEGF, KDR e Flt-1 entre os grupos e nas idades gestacionais analisadas, aos 30 dias notou-se uma tendência entre os grupos em relação a expressão do Flt-1. Os fatores de crescimento b FGF, FGFR-1 e FGFR-2, não apresentaram diferenças em suas expressões, somente aos 35 dias de gestação para o mRNA FGFR-1. Conclui-se que o saco vitelino está intensamente relacionado com o desenvolvimento embrionário, na qual sua involução e desaparecimento relacionam-se com o crescimento e \"maturação\" embrionária. / High mortality rates occur during embryonic implantation embryos from IVF and FTAI. It is believed that the failure in the development of the yolk sac may be related to these embryonic losses. The objective of this study was to describe the macroscopic, microscopic and mRNA expression features of the yolk sac involution process, since the literature relates contradictory information regarding the processes of formation and involution of the yolk sac in IVF and TAI embryos. Nineteen embryos were used for FTAI and 16 IVF; the specimes were obtained from the Sociedade Agropecuária Imaculada Conceição-Redenção/PA, Brazil. No significant differences were observed in the size of embryos aged 25 (p = 0.1), 35 (p = 0.1), 40 (p = 0.4) and 45 (p = 0.36) days in the groups of IVF and FTAI. However between FTAI and IVF embryos with 30 days of gestation, a difference (p = 0.03) was detectable. The growth curve (Crow-Rump) increased gradually and linearly with gestational age. Macroscopically, it was observed that after 25 days of gestation, the yolk sac was \"T-shaped\" and transparent colored. A central portion was observed in the vicinity of the ventral ends of the embryo exhibiting two thin membranes, which accompanied the allantois with the umbilical vessels. At 30 to 35 days this portion becomes yellowish and its central part together with the extremities was reduced displaying clearly signs of regression. At 40 to 45 days, the process of involution was presented in a curled fashion and ends with the central region resembling a flat small grain in contact with the amnion. Microscopically, the yolk sac was formed by three layers: the endoderm, single layer lining the vitelline cavity, an intermediate layer and vascular mesenchymal mesothelium, exoceloma focused on single layer. The epithelium of the yolk sac formed folds or crypts, which projected to the light. Immunohistochemistry marking was noted for antibodies VEGF, Flt-1 and KDR. No significant difference were observed in mRNA expression of VEGF, KDR and Flt-1 between groups and the gestational ages analyzed at 30 days there has been a trend among the groups regarding the expression of Flt-1. The growth factors bFGF, FGFR-1 and FGFR-2 showed no differences in their expressions, with the exception of 35 days of gestation for the m-RNA FGFR 1. It was concluded that the yolk sac is strongly related to embryonic development, in which the involution and disappearance are related to growth and \"maturity\" stage.

A inseminação artificial em tempo fixo (IATF), juntamente com a inseminação artificial pós-cervical (IAPC), permite uma redução expressiva da mão de obra e uma melhor utilização dos machos geneticamente superiores. O objetivo deste estudo foi avaliar a eficiência do eCG associado agonista de GnRH (buserelina) ou apenas buserelina para indução e sincronização da ovulação em porcas desmamadas e submetidos a apenas uma IATF. Um total de 495 femeas suínas desmamadas foram utilizadas, cuja detecção de estro foi realizada uma vez ao dia (07:30). As fêmeas foram divididas em três tratamentos: Controle (n = 165) - a primeira IA foi realizada no início do estro (0 h) e repetida a cada 24 h, posteriormente, durante o estro; Tratamento 2: eCG + GnRH (n = 165) – femeas receberam uma injeção intramuscular de 600 UI de eCG após o desmame, e uma injeção intramuscular de buserelina (10 μg) 86-89 h após a administração de eCG; Tratamento 3: GnRH - fêmeas receberam uma injeção intramuscular de buserelina (10 μg) 86-89 h após o desmame. Nos três grupos era realizada a exposição ao macho no dia seguinte ao desmame uma vez ao dia, e aquelas fêmeas que demostrassem estro eram retiradas dos protocolos de IATF. Os grupos tratados receberam uma única IA, 118-120 h após o desmame (30-33 h após a buserelina). A IAPC contou com doses homospérmicas (1,5 x 109 células de espermáticas/50 ml) que foram utilizadas nas femeas em todos os três tratamentos. Não houve diferenças entre os tratamentos quanto ao número de femeas em estro até o terceiro dia (P>0,05). O intervalo entre o desmame e a ovulação foi significativamente maior (P< 0,05) no grupo controle (141,5 ± 1,58 h) comparativamente aos tratamentos eCG+GnRH (133,3 ± 1,60 h) e GnRH (135,9 ± 1,57 h). As femeas do tratamento eCG+GnRH ovularam em média antes (P<0,05) em relação ao grupo GnRH (44,5 ± 1,74 vs 48,2 ± 1,73 h). A taxa de parto foi superior (P<0,05) no grupo de controle, comparativamente aos tratamentos eCG + GnRH e GnRH, mas não houve diferença (P>0,05) em leitões nascidos totais entre os tratamentos. O uso do GnRH, com ou sem a administração prévia de eCG, antecipa a ovulação nas fêmeas suínas desmamadas. Nas fêmeas tratadas com eCG+GnRH ou GnRH, há em maior percentual de fêmeas inseminadas fora do momento ideal, em relação à ovulação, o que pode prejudicar o desempenho reprodutivo, sendo necessários ajustes no momento de aplicação dos hormônios ou no momento da inseminação para estudos subsequentes. / Fixed-time artificial insemination (FTAI) together with post-cervical artificial insemination (PCAI), allows an expressive reduction in labor requirements and a wider use of higher indexing boars. The aim of this study was to evaluate the efficiency of eCG agonist of GnRH (buserelin) or just buserelin for induction and synchronization of ovulation in weaned sows and submitted to FTAI .A total of 495 weaned sows whose estrus detection was performed once daily (07:30 A.M), starting a day after weaning. The sows were allocated into three treatment groups: Control (n=165) – the first AI was performed at estrus onset (0 h) and repeated every 24 h thereafter during estrus; Treatment 2: eCG + GnRH (n=165) – sows received an intramuscular injection of 600 UI eCG after weaning, and an intramuscular injection of buserelin (10 μg) 86-89 h after eCG administration; Treatment 3: GnRH - sows received an intramuscular injection of buserelina (10 μg) 86-89 h after weaning. . In the three groups it was performed boar exposure in the following day to weaning once a day, and those sows that demonstrated oestrus were removed from FTAI protocols. Treated groups received a single AI, 118-120 h after weaning (30-33 h after buserelin). PCAI with homospermic doses (1.5 x 109 total of sperm cells/50 ml) were respectively performed in sows in all treatment group. There were no differences (P> 0,05) in number of sows that showed oestrus until the third day between treatments. The interval between weaning and ovulation was significantly higher (P< 0,05) in the control group (141,5 ± 1,58 h) comparatively the treatments eCG+GnRH (133,3 ± 1,60 h) and GnRH (135,9 ± 1,57 h). Sows of treatment eCG+GnRH ovulated earlier (P< 0,05) comparing to the GnRH group (44,5 ± 1,74 vs 48,2 ± 1,73 h). Higher (P<0,05) farrowing rate in control group comparatively to treatments eCG+GnRH e GnRH was observed, but there were no differences (P> 0,05) in total piglets born between treatments. Use of GnRH, with or without previous administration of eCG anticipates ovulation in sows weaned, in the sows treated with eCG + GnRH or GnRH, there is a greater percentage of females inseminated outside the ideal time, with respect to ovulation, it can harm the reproductive performance, with necessary adjustments at the time of application hormones or at the time of insemination for subsequent studies.

Un groupe de robots collaboratifs peut gérer des tâches qui sont difficiles, voire impossibles, à accomplir par un seul. On appelle un ensemble de robots coopérant un système multi-agents (SMA). L'interaction entre agents est un facteur clé dans la commande coopérative qui pose d'importants défis théoriques et pratiques. L'une des tâches du contrôle coopératif est le consensus dont l'objectif est de concevoir des protocoles de commande afin de parvenir à un accord entre leurs états respectifs. Cette thèse améliore la navigation pour les SMA, tout en tenant compte de certaines contraintes pratiques (modèle du robot et contraintes temporelles) dans la conception de contrôleurs coopératifs pour chaque agent, de manière décentralisée. Dans cette thèse, deux directions sont étudiées. D'une part, le taux de convergence est une spécification de performance importante pour la conception du contrôleur pour un système dynamique. La convergence rapide est toujours recherchée pour améliorer les performances et la robustesse. La plupart des algorithmes de consensus existants se concentrent sur la convergence asymptotique, où le temps d'établissement est infini. Cependant, de nombreuses applications nécessitent une convergence rapide généralement caractérisée par une stratégie de commande à temps fini. De plus, la commande à temps fini autorise certaines propriétés intéressantes, mais le temps de stabilisation dépend des conditions initiales des agents. L'objectif ici est de concevoir un protocole de consensus leader-follower à temps fixe pour les SMA décrits en temps continu. Ce problème est étudié en utilisant la théorie de la stabilisation à temps fixe, qui garantit que le temps de stabilisation est borné quelles que soient les conditions initiales. Les contrôleurs et les observateurs à modes glissants sont conçus pour que chaque agent résolve le problème du consensus à temps fixe lorsque le leader est dynamique. D'autre part, par rapport aux systèmes à temps continu, le problème du consensus dans un cadre à temps discret convient mieux aux applications pratiques en raison de la limitation des ressources de calcul pour chaque agent. Le modèle de commande prédictive (MPC) permet de gérer les contraintes de commande et d'état des systèmes. Dans cette thèse, cette méthode est appliquée pour traiter le problème du consensus en temps discret en laissant chaque agent résoudre, à chaque étape, un problème de commande optimale contraint impliquant uniquement l'état des agents voisins. Les performances de suivi sont également améliorées dans cette thèse en ajoutant de nouveaux termes à partir du MPC classique. Les contrôleurs proposés sont simulés et implémentés sur un groupe composé de plusieurs robots réels en utilisant ROS (Robotic Operating System). Dans cette thèse, quelques solutions correspondant au problème de la connexion entre plusieurs robots mobiles de manière décentralisée, du réglage des périodes d'échantillonnage et des paramètres de contrôle sont également abordées. / Nowadays, robots have become increasingly important to investigate hazardous and dangerous environments. A group of collaborating robots can often deal with tasks that are difficult, or even impossible, to be accomplished by a single robot. Multiple robots working in a cooperative manner is called as a Multi-Agent System (MAS). The interaction between agents to achieve a global task is a key in cooperative control. Cooperative control of MASs poses significant theoretical and practical challenges. One of the fundamental topics in cooperative control is the consensus where the objective is to design control protocols between agents to achieve a state agreement. This thesis improves the navigation scheme for MASs, while taking into account some practical constraints (robot model and temporal constraints) in the design of cooperative controllers for each agent, in a fully decentralized way. In this thesis, two directions are investigated. On one hand, the convergence rate is an important performance specification to design the controller for a dynamical system. As an important performance measure for the coordination control of MASs, fast convergence is always pursued to achieve better performance and robustness. Most of the existing consensus algorithms focus on asymptotic convergence, where the settling time is infinite. However, many applications require a high speed convergence generally characterized by a finite-time control strategy. Moreover, finite-time control allows some advantageous properties but the settling time depend on the initial states of agents. The objective here is to design a fixed-time leader-follower consensus protocol for MASs described in continuous-time. This problem is studied using the powerful theory of fixed-time stabilization, which guarantee that the settling time is upper bounded regardless to the initial conditions. Sliding mode controllers and sliding mode observers are designed for each agent to solve the fixed-time consensus tracking problem when the leader is dynamic. On the other hand, compared with continuous-time systems, consensus problem in a discrete-time framework is more suitable for practical applications due to the limitation of computational resources for each agent. Model Predictive Control (MPC) has the ability to handle control and state constraints for discrete-time systems. In this thesis, this method is applied to deal with the consensus problem in discrete-time by letting each agent to solve, at each step, a constrained optimal control problem involving only the state of neighboring agents. The tracking performances are also improved in this thesis by adding new terms in the classical MPC technique. The proposed controllers will be simulated and implemented on a team of multiple Mini-Lab Enova Robots using ROS (Robotic Operating System) which is an operating system for mobile robots. ROS provides not only standard operating system services but also high-level functionalities. In this thesis, some solutions corresponding to problem of connection between multiple mobile robots in a decentralized way for a wireless robotic network, of tuning of the sampling periods and control parameters are also discussed.

<p dir="ltr">Nulliparous replacement beef heifers represent an opportunity to improve both genetic potential and lifetime production within the herd. However, advances in reproductive efficiency and synchrony in the herd require heifers to conceive earlier than multiparous cows in order to account for the extended postpartum interval following first-time parturition. Further, those heifers that achieve early calving dates continue to calve early throughout their lifetime. This early calving results in increased weaning weights and better synchrony in subsequent years. One obstacle producers face when breeding nulliparous heifers is the ability to identify which animals have reached puberty at the initiation of the breeding season. This results in the variation observed in synchronization success within a group of heifers. In Chapter 2 our laboratory formulated a study to analyze the efficacy of the 7-d CO-synch + CIDR protocol when utilizing short-term exposure to a progestin (melengestrol acetate, MGA) as a pre-synchronization protocol. Our hypothesis was that the heifers may respond more efficiently to an orally fed progestin as the increase in serum progesterone is less pronounced, when compared to the CIDR. This first study resulted in a protocol application error, in which MGA was fed an extra day (8 vs. 7). This additional day of treatment with MGA following the administration of a prostaglandin eliminated our laboratory’s opportunity to collect meaningful data from this first attempt. Thus, it was pertinent to restart the experiment in an attempt to obtain data for analysis. In order to avoid confounding data, half of the heifers in each treatment received a new treatment, while the other half remained on the treatment assigned during the first attempt. After completing the second round of experiments it was observed that at the first pregnancy check (study day 30), the heifers that were originally assigned to MGA, but were switched to a CIDR for the second attempt numerically outperformed their counterparts in the other three formulated treatment groups. In an attempt to replicate these results, a second study was formulated (Chapter 3), to mimic the timeline from the preliminary study. This resulted in a pre-synchronization treatment protocol prior to the initiation of the breeding season synchronization protocol. Previous studies that have been conducted with similar protocols were designed to provide heifers with a pre-synchronization period that would aid in the attainment of puberty prior to their first attempt at breeding through exposure to progesterone. The purpose of the Chapter 3 experiment was to evaluate the effects of feeding melengestrol acetate (MGA®) as a pre-synchronization for 10 days immediately prior to estrous synchronization and fixed-timed artificial insemination (FTAI). The ten days were chosen as this was a novel protocol that was the result of the preliminary study and the subsequent restart. Ninety-three crossbred heifers (395.67 ± 5.37 kg) were blocked by BW, genetics, and reproductive tract scores and allotted to 3 treatments. The 3 treatment groups were: 7-day CO-synch + CIDR without pre-synchronization (CON, n=31); 25 mg PGF2α (Lutalyse®) followed immediately by MGA feeding for 7 d prior to the start of the 7-day CO-synch + CIDR program (PRE, n=31); and 25 mg PGF2α followed immediately by MGA feeding for 7d followed 10 d later by the start of the 7-day CO-synch + CIDR program (PRE+10, n=31). The 7-day Co-synch + CIDR protocol in all three treatments was initiated on d 0 by administering a 2 cc IM injection of GnRH (Cystorelin®) and placing a CIDR into the vagina. The CIDRs were removed 7 days later and accompanied by 25 mg IM injection of PGF2α. An injection of GnRH occurred 60-66 hours following PGF2α at FTAI with frozen semen from a single bull. Ten days after FTAI, heifers were exposed to a bull. Estrotect® patches were applied throughout the study to assess estrous behavior. Ovaries were visualized by transrectal ultrasonography 24 hours post-FTAI to determine whether ovulation occurred. Pregnancy was determined on days 40, 64, and 109 post-FTAI via transrectal ultrasonography. Blood was collected via jugular venipuncture (d -19 and -12, MGA initiation and termination, days 0, 7, and 9) and serum progesterone determined. Performance data were analyzed using the MIXED procedure and conception data were analyzed using the GLIMMIX procedure of SAS. Heifers in the PRE+10 treatment group had higher levels of progesterone (P=0.04) at d 0 compared to PRE heifers. At d 7 (CIDR removal) there was a tendency (P=0.07) for PRE+10 heifers to have higher levels of progesterone than PRE, but did not differ by d 9 (FTAI, P=0.36). FTAI conception rates in heifers in the PRE treatment group (63%) tended (P = 0.09) to be higher compared to the controls (35%), but not different from PRE+10 (43%), with no differences in season-long pregnancy rates (P > 0.15). Pre-synchronization with MGA immediately prior to FTAI synchronization appears to increase conception rates early in the breeding season in beef heifers. In Chapter 4, our laboratory analyzed the efficacy of supplementation strategies post-insemination. Unlike the first two experiments, the third study focused on reproductive failure that occurs after insemination. The overarching goal remained the same, increasing the reproductive efficiency within our nulliparous heifer herd. A common practice for beef producers in the United States is to use estrous synchronization, and immediately turn heifers out to lush spring pasture immediately following FTAI. The fresh forage is high in water content which lowers dry matter intake (DMI) and creates a negative energy balance. Ultimately, this reduced energy intake can result in a reduction in reproductive performance. In the Chapter 4 our laboratory formulated a supplementation strategy utilizing the SmartFeed™ technology to deliver soybean hulls to our treatment group following insemination for 45 days. The utilization of the SmartFeed® technology provided the opportunity to analyze the efficacy of the supplementation strategy using each individual animal as an experimental unit. Sixty-two nulliparous crossbred heifers were fed in drylot to obtain moderate body condition prior to breeding at d 0 via FTAI and trained to utilize the SmartFeed™ system.Heifers were blocked by weight and body condition score to either the soybean hull supplementation group (SOY) or the non-supplemented control group (CON). . Heifers in the treatment group received their supplementation by entering the SmartFeed™ system, allowing for RFID controlled release of 2.27 kg. per head each day for 45 days. Scales located beneath each feed pan sent real time weight data for regulation and analysis on individual animal feeding behavior. On study d 6, a subset of the nulliparous crossbred heifers (n = 12; n = 6/ treatment) were transported by trailer from the Feldun Purdue Agricultural Center to Purdue West Lafayette main campus (approximately 217.74 km.). The subset of heifers had embryos flushed and evaluated for embryo quality and number of live/dead cells. Ultrasonography was utilized to monitor ovarian activity throughout the duration of the study and to determine pregnancy status 30 days after FTAI and 30 days after the 45-day breeding season. Though conception rates were not statistically (P=0.17) different (SOY 16/25; 64% vs. CON 11/25; 44%) when comparing treatment groups, the numerical differences suggest there is potential in pursuing a similar supplementation strategy following breeding in nulliparous beef heifers. The supplementation of SBH resulted in greater weight gain over time (P = 0.04), potentially explaining the numeric improvement in conception rate. The two pre-synchronization studies from Chapters 2 and 3 resulted in numerical improvements in conception rate as a result of exposing replacement heifers to a source of progestin prior to the initiation of their synchronization protocol. The implementation of progesterone priming mitigates the occurrence of short cycles and immature oocyte maturation at the time of ovulation. Based on results from previous studies conducted by our laboratory, the source of progestin greatly determines the timing and concentration of P4 circulating in replacement heifers immediately following treatment. Additionally, the 7 & 7 estrous synchronization protocol has grown in popularity and implementation. This protocol when broken down is very similar to the pre-synchronization protocol our laboratory utilized for the first two studies. Several studies have been conducted to analyze the efficacy of the 7 & 7 and the conception data was comparable to the MGA derived protocol. Thus, one potential direction for the future would be to formulate a research study that compares the two protocols. Since the conception rates were similar, it may be hypothesized that the MGA protocol could be more widely accepted as there is a reduction number of times animals are handled.</p>

L’objectif principal de cette thèse est d’étudier le problème du contrôle de suivi distribué pour les systèmes de formation de multi-robots à contrainte non holonomique. Ce contrôle vise à entrainer une équipe de robots mobile de type monocycle pour former une configuration de formation désirée avec son centroïde se déplaçant avec une autre trajectoire de référence dynamique et pouvant être spécifié par le leader virtuel ou humain. Le problème du contrôle de suivi a été résolu au cours de cette thèse en développant divers contrôleurs distribués pratiques avec la considération d’un taux de convergence plus rapide, une précision de contrôle plus élevée, une robustesse plus forte, une estimation du temps de convergence explicite et indépendante et moins de coût de communication et de consommation d’énergie. Dans la première partie de la thèse nous étudions d’abord au niveau du chapitre 2 la stabilité à temps fini pour les systèmes de formation de multi-robots. Une nouvelle classe de contrôleur à temps fini est proposée dans le chapitre 3, également appelé contrôleur à temps fixe. Nous étudions les systèmes dynamiques de suivi de formation de multi-robots non holonomiques dans le chapitre 4. Dans la deuxième partie, nous étudions d'abord le mécanisme de communication et de contrôle déclenché par l'événement sur les systèmes de suivi de la formation de multi-robots non-holonomes au chapitre 5. De plus, afin de développer un schéma d'implémentation numérique, nous proposons une autre classe de contrôleurs périodiques déclenchés par un événement basé sur un observateur à temps fixe dans le chapitre 6. / The main aim of this thesis is to study the distributed tracking control problem for the multi-robot formation systems with nonholonomic constraint, of which the control objective it to drive a team of unicycle-type mobile robots to form one desired formation configuration with its centroid moving along with another dynamic reference trajectory, which can be specified by the virtual leader or human. We consider several problems in this point, ranging from finite-time stability andfixed-time stability, event-triggered communication and control mechanism, kinematics and dynamics, continuous-time systems and hybrid systems. The tracking control problem has been solved in this thesis via developing diverse practical distributed controller with the consideration of faster convergence rate, higher control accuracy, stronger robustness, explicit and independent convergence time estimate, less communication cost and energy consumption.In the first part of the thesis, we first study the finite-time stability for the multi-robot formation systems in Chapter 2. To improve the pior results, a novel class of finite-time controller is further proposed in Chapter 3, which is also called fixed-time controller. The dynamics of nonholonomic multi-robot formation systems is considered in Chapter 4. In the second part, we first investigate the event-triggered communication and control mechanism on the nonholonomic multi-robot formation tracking systems in Chapter 5. Moreover, in order to develop a digital implement scheme, we propose another class of periodic event-triggered controller based on fixed-time observer in Chapter 6.

La intersección de las Avenidas José Granda y Universitaria, ubicado en el distrito de San Martín de Porres, Lima (conocido como Óvalo José Granda); es un cruce con un diseño geométrico no convencional, que tiene forma de rotonda, pero que no opera como tal; consta de un cruce semaforizado de cuatro accesos que cruzan la rotonda al interior de la misma, en los sentidos Norte-Sur, Sur-Norte, generando entrecruzamientos en todos los carriles de la rotonda y muy bajas velocidades, que son sumamente peligrosos en su operación. Para, mitigar el problema encontrado, se ha realizado el rediseño de la intersección implementando un intercambio vial semaforizado tipo diamante a nivel; para el cual se han aplicado dos alternativas de coordinación semafórica, ambas con control de tiempo fijo. De la modelación del tránsito vehicular en los software AIMSUN para el análisis de microsimulación y SYNCHRO para la optimización en la coordinación semafórica; de los tres escenarios evaluados (situación actual, alternativas 1 y 2), se compararon dos medidas de efectividad, el tiempo de demoras (seg/veh) y el número de paradas (#/veh/km), concluyendo que los resultados operacionales mejoran con la alternativa 1; sin embargo, empeoran con la alternativa 2, sometidas a evaluación contra la situación actual. Se recomienda analizar las alternativas planteadas con semaforización actuada (de tiempo flexible, activado por el tránsito); así como también, con paraderos de transporte público, se ubiquen fuera el área de análisis, para que no afecte la operación del diseño propuesto. / The intersection of José Granda and Universitaria avenues, located in the district of San Martín de Porres, Lima (known as Óvalo José Granda), is a junction with an unconventional geometric design, which is shaped like a roundabout, but does not operate as such. It consists of a traffic light crossing with four accesses that cross the roundabout inside it, in the North-South, South-North directions, generating intersections in all the lanes of the roundabout and very low speeds, which are extremely dangerous in its operation. To mitigate the problem encountered, the intersection has been redesigned by implementing a diamond-type traffic light interchange, at level; for which two alternatives of traffic light coordination have been applied, both with fixed time control. From vehicle traffic modeling in AIMSUN software for microsimulation analysis and SYNCHRO for traffic light coordination optimization; Of the three evaluated scenarios (current situation, alternatives 1 and 2), two measures of effectiveness were compared, the delay time (sec / veh) and the number of stops (# / veh / km), concluding that the operational results improve with alternative 1; however, they worsen with alternative 2, evaluated against the current situation. It is recommended to analyze the alternatives proposed with activated traffic lights (flexible time, activated by traffic); as well as, with public transport stops, are located outside the analysis area, so that they do not affect the operation of the proposed design. / Tesis