Orientador: Fosca P. Pereira Leite / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-07-19T22:55:07Z (GMT). No. of bitstreams: 1 Arantes_IsabelCristina_M.pdf: 8344654 bytes, checksum: d646b8f7e725dc5c2819b61352bc3d45 (MD5) Previous issue date: 1994 / Resumo: Os ermitões utilizam conchas vazias de gastrópodes para protegerem o frágil abdômen devido à falta de calcificação. Essa associação pode influenciar vários aspectos da sua biologia. Foram feitas amostragens mensais durante o ano de 1990 na região do Araçá, em frente à Ilha de Pernambuco (23º48'57¿S e 45º24'27"W), para constatar as associações entre as espécies de ermitão e as conchas de gastrópodes. Quatro espécies de ermitão foram encontradas, Clibanarius vittatus, C. sclopetarius, C. antillensis e Pagurus criniticomis. Para Clibanarius vittatus, por ser a espécie mais abundante, foram estudadas as relações entre as características morfométricas das conchas com o tamanho do animal e, em laboratório, observou-se a preferência por conchas de diferentes tamanhos e espécies. Todos os animais coletados foram medidos e diferenciados quanto ao sexo, e as respectivas conchas analisadas quanto ao comprimento total e abertura, peso e volume interno. As relações entre o comprimento total, peso e abertura das conchas e o tamanho de C. vittatus foram significativas. Machos e fêmeas ocupavam conchas de Siratus senegalensis e Cymatium parthenopeum semelhantes quanto ao comprimento e peso. Entretanto, quando utilizavam conchas de Stramonita haemastoma, as fêmeas ocupavam as maiores e mais pesadas. Em laboratório, os machos apresentaram preferências por conchas maiores que as das fêmeas. Estas apresentaram preferência por conchas de Cymatium, enquanto que no ambiente ocupavam especialmente conchas de Siratus. A oferta de conchas na natureza pode não estar suprindo as necessidades de conchas de tamanhos ideais para o macho, mas pode ser satisfatória às fêmeas. As preferências das fêmeas podem estar relacionadas com o custo energético de carregar uma concha pesada, além do peso dos ovos, durante a fase reprodutiva / Abstract: Hermit crabs utilize empty gastropod shells to protect their vulnerable abdomen because of the lack of calcification. This association of crabs, and shells can influence many aspects of crab biology. Samples were collected monthly during 1990 at Araçá region, in front of Pernambuco island (23º48'57"S e 45º24'27"W), to verify the relationship between hermit crab, and shell species. Four hermit crab species were found, Clibanarius vittatus, C. sclopetarius, C. antillensis and Paguros criniticornis. For Clibanarius vittatus, the most common species, the relationship between shell character morphometrics, and animal size was studied. C. vittatus's preference for shells of different species, and sizes was also observed in laboratory. All the animals were measured, and sexed, and shells' attributes, such as total length, aperture length, weight, and internal volume were measured. The relationship between animal size, and total length, aperture lentgh, and weight of the shells were significant for C. vittatus. Shell utilization patterns in nature varied between sexes and shell species. Males and females utilized similar shells of Siratus senegalensis and Cymatium parthenopeum, for size and weight. However, the shells of Stramonita haemastoma were used, females occupied the bigger and heavier ones. In laboratory, males showed preference for bigger shells, where as females preferred shells of Cymatium although in nature they occupied mainly Siratus. The availability of shells in the environment may be not sufficient for the males, generally bigger than females. On the other hand, for females shells seem relatively available. The energetic costs of carrying a heavy shell (besides the eggs, during the reproductive phase) may determine the preference of females / Mestrado / Mestre em Ciências Biológicas

Orientador: Darlene Aparecida de Oliveira Ferreira / Banca: Izabel de Carvalho / Banca: Leila de Menezes Stein / Resumo: A dissertação que apresentamos traz como questão principal o problema da caracterização social dos grupos de produtores rurais que controlam (mesmo que parcialmente) os próprios meios de produção. O estudo se divide em duas partes: a primeira, um debate teórico em que discutimos as diversas nominações atribuídas a esses agricultores ao longo da literatura nacional, procurando estabelecer uma definição coerente para cada uma delas, bem como suas implicações teóricas e políticas na análise do mundo rural. O conceito de agricultura familiar emergiu da discussão teórica como a maneira mais adequada de se definir o grupo social em questão. Na segunda parte, realizou-se um estudo empírico tomando como base o município de Conchas, interior de São Paulo, no intuito de testar até que ponto a definição adotada correspondia às características dos agricultores observadas em campo. / Abstract: The papert we present brings as ultimate issue the problem of the social characterization of the groups of agricultural producers that controls (fully or partially) the means of production. The study is divided in two parts: the first one, a theoretical quarrel where we discuss the diverse nominations attributed to these agriculturists throughout national literature, looking for to establish a coherent definition for each one of these nominations, as well as its implications, theoreticians and politics in the analysis of the agricultural world. The concept of family farm business emerged of the theoretical quarrel as the adjusted way to define the social group in question. In the second part, an empirical study was become fulfilled taking as base the town of Conchas, at the countryside of São Paulo state, in intention to test the adopted definition to see if it matches to the characteristics of the agriculturists observed in field. / Mestre

Made available in DSpace on 2014-06-11T19:29:47Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-04-11Bitstream added on 2014-06-13T21:00:13Z : No. of bitstreams: 1 volpi_gc_me_arafcl.pdf: 1655826 bytes, checksum: 4b25b334015c6264ee03e76bcd004257 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A dissertação que apresentamos traz como questão principal o problema da caracterização social dos grupos de produtores rurais que controlam (mesmo que parcialmente) os próprios meios de produção. O estudo se divide em duas partes: a primeira, um debate teórico em que discutimos as diversas nominações atribuídas a esses agricultores ao longo da literatura nacional, procurando estabelecer uma definição coerente para cada uma delas, bem como suas implicações teóricas e políticas na análise do mundo rural. O conceito de agricultura familiar emergiu da discussão teórica como a maneira mais adequada de se definir o grupo social em questão. Na segunda parte, realizou-se um estudo empírico tomando como base o município de Conchas, interior de São Paulo, no intuito de testar até que ponto a definição adotada correspondia às características dos agricultores observadas em campo. / The papert we present brings as ultimate issue the problem of the social characterization of the groups of agricultural producers that controls (fully or partially) the means of production. The study is divided in two parts: the first one, a theoretical quarrel where we discuss the diverse nominations attributed to these agriculturists throughout national literature, looking for to establish a coherent definition for each one of these nominations, as well as its implications, theoreticians and politics in the analysis of the agricultural world. The concept of family farm business emerged of the theoretical quarrel as the adjusted way to define the social group in question. In the second part, an empirical study was become fulfilled taking as base the town of Conchas, at the countryside of São Paulo state, in intention to test the adopted definition to see if it matches to the characteristics of the agriculturists observed in field.

El modelo de biomasa dinámica de Schaefer fue usado para evaluar el estado de la pesquería del recurso “concha negra” (Anadara tuberculosa y Anadara similis), usando los datos de captura, esfuerzo y CPUE procedentes de los registros del IMARPE de los manglares de Puerto Pizarro, durante el periodo 1996 - 2010. El modelo aplicado es resuelto por error de proceso, que es uno de los más utilizados para estimar los parámetros de los modelos cuantitativos, el proceso de ajuste se hizo mediante la suma de la máxima verosimilitud, el cual presentó un buen ajuste del modelo en función a las características del recurso. La incertidumbre de los parámetros del modelo y las variables derivadas fueron obtenidas mediante el bootstrap no-paramétrico. Los parámetros poblacionales estimados fueron: K: 66036 kg, r: 0.23 kg*año-1 y q: 0.000125, con esta base se calcularon los principales puntos de referencia RMS: 3806 kg ERMS: 1489 viajes y FRMS: 0.12. Así como, la relación entre la biomasa actual y la biomasa prístina B2010/K: 0.2, B2010/B1996: 0.41 y la relación entre la mortalidad por pesca actual y la mortalidad por pesca al RMS F/FRMS: 2. De acuerdo con el análisis de estos resultados el recurso se encuentra sobreexplotado, encontrándose el 74% de la biomasa por debajo de aquella del máximo rendimiento sostenible. Las proyecciones de la biomasa sugieren que el nivel de explotación no debe sobrepasar los 1000 kg para evitar el colapso de la pesquería. Palabras claves: Concha negra, Evaluación, Modelo de biomasa dinámica, Manglar, Tumbes. / --- The model of Schaefer biomass dynamics was used to assessement the status of the fishery for Anadara tuberculosa y Anadara similis as a single resource black shell. Catch, effort and CPUE data from IMARPE registered in Port Pizarro for the period 1996-2010. The model is solved by process error, which is one of the most used for estimating the parameters of quantitative models; the adjustment was made by maximum likelihood, which presents a good fit of the model according to the biology of the resource. The uncertainty of the model parameters and derived variables were obtained by nonparametric bootstrap. The parameter of the biomass dynamics model were: K: 66036 kg, r: 0.23 kg*year-1 and q: 0.000125, the estimation of the point reference were: RMS: 3806 kg, ERMS: 1489 tasks y FRMS: 0.12. Also the relationship between B2010/K: 0.2, B2010/B1996:0.41 and F2010/FRMS: 2 to make a diagnosis of the resource status. The results of the analysis indicate that the resource is overexploited, found 74% of the biomass at maximum sustainable yield, the biomass projections suggest that the exploitation rate should not exceed 1000 kg to avoid collapse of the fishery. Keywords: black shell, Assessment, Biomass dynamics model, Mangrove, Tumbes.

Made available in DSpace on 2018-08-01T23:30:06Z (GMT). No. of bitstreams: 1 tese_10143_mestrado final CORRIGIDO Adriana.pdf: 4344368 bytes, checksum: 7247e25bdbd385877f17456e194bcc8b (MD5) Previous issue date: 2016-08-09 / Esta pesquisa visa compreender as relações entre o artesanato, o patrimônio cultural imaterial, a memória coletiva e a identidade, analisando as influências que um exerce sobre o outro. Para tanto, realizamos um estudo de campo entre os artesãos e moradores de Piúma, pois o ofício é não registrado como patrimônio cultural imaterial. Sendo assim, analisamos os modos de fazer do grupo, as semelhanças e diferenças em relação aos demais municípios que também produzem artesanato de conchas, bem como as relações, sociais e políticas envolvidas Inicialmente apresentamos o referencial teórico utilizado para a análise dos dados coletados e as discussões acerca da memória coletiva. Em seguida, observamos a trajetória histórica do artesanato de conchas depois da década de 1960, as principais diferenças existentes no município de Piúma e empreenderemos a discussão de Patrimônio Cultural, baseada no modo de enxergar dos artesãos e moradores. Por fim, analisaremos as peças e a relação com a identidade construída a partir do artesanato de conchas.

El presente trabajo de tesis tiene como objetivo identificar y cuantificar las diferentes alteraciones morfológicas de espermatozoides de Argopecten purpuratus, ocasionadas por el proceso de criopreservación, y relacionarlas con su motilidad y capacidad de fecundación post-descongelamiento. Se utilizaron conchas de abanico maduras obtenidas de cultivos suspendidos de la Bahía la Herradura de Guayacán, Coquimbo-Chile. Se disectó la gónada y la porción masculina se cortó en trozos para liberar los espermatozoides en AMFE. Estos, fueron incubados en una solución crioprotectora compuesta por ME2SO al 10%, 125 mM de sacarosa y 10% de vitelo del huevo de gallina. Se congeló los espermatozoides a una tasa de 8,8 ºC/min. El descongelamiento se realizó 24 horas después mediante inmersión en un recipiente con agua a 50º C durante 20 seg y luego en otro a temperatura ambiente. Posteriormente se cuantificó el número de espermatozoides móviles en un microscopio de luz; también se cuantificó la cantidad de espermatozoides lesionados en la cabeza, acrosoma, pieza media y flagelo en un microscopio electrónico de barrido. Finalmente, se determinó la capacidad fecundante de los espermatozoides inseminando ovocitos frescos. La motilidad varió significativamente entre los tratamientos (C, PRE y POST) en cuanto a porcentaje de espermatozoides móviles y tiempo de actividad. Se observó distintas alteraciones en la morfología del espermatozoide congelado-descongelado. Algunas tuvieron la cabeza deforme, hinchada o lisada y la membrana celular plegada o rota. También fue observada, reacción acrosómica y mitocondrias fuera de su posición normal o ausentes. Pero las alteraciones más comunes se encontraron en el flagelo, el cual sufrió ruptura, rigidez y pérdida de su estructura lineal. En C y PRE el porcentaje de espermatozoides ilesos fue mayor (87,7 ± 3,4 % y 79,0 ± 3,0 % respectivamente), mientras que en POST fue 14,2 ± 2,8 %. La estructura lesionada en la mayor cantidad de espermatozoides POST, fue el flagelo (77,0 ± 3,0 %); el porcentaje de espermatozoides con lesión en la cabeza fue 55,1 ± 7,4 % y los que tuvieron el acrosoma reaccionado fueron 28,7 ± 3,3 %. La pieza media fue afectada en el 23,9 ± 4,1 % de espermatozoides. Los porcentajes de fecundación fueron 68,3 ± 6,6 %, 67,9 ± 4,2 % y 58,2 ± 7,3 % para C, PRE y POST respectivamente, no encontrándose diferencias significativas entre ellos. Se correlacionó la motilidad total cuantificada a los 5 y a los 30 minutos, las lesiones ocurridas en las diferentes estructuras espermáticas y los porcentajes de fecundación; encontrándose mayor correlación entre las alteraciones y la motilidad que entre las alteraciones y los porcentajes de fecundación. La correlación entre la motilidad y fecundación fue baja (0,650 y 0,668 con la motilidad a los 5 y 30 min respectivamente). La incubación durante el tiempo de equilibrio en la solución crioprotectora utilizada, no tiene un efecto tóxico que altere la motilidad o la morfología de los espermatozoides, sino más bien, tiene un efecto activador de la motilidad. Esta propiedad de la solución crioprotectora, se suma a la de protección, que les daría a los espermatozoides durante el congelamiento-descongelamiento, que es la etapa del proceso de criopreservación durante la cual los espermatozoide son afectado negativamente.. Las cabezas destrozadas de los espermatozoides sería consecuencia de la lisis celular o del crecimiento de cristales de hielo, todo esto como consecuencia de desórdenes osmóticos. La rigidez de los flagelos podría ser por alteraciones en la estructura del axonema. Posiblemente, la reacción acrosomal sería ocasionada por los cambios de polaridad ocurridos por el congelamiento-descongelamiento. En general, las lesiones en cualquiera de las estructuras espermáticas están relacionadas finalmente con la membrana celular. / The present work identifies and quantifies the morphological external alterations of spermatozoa of the scallop A. purpuratus due to long-term cryopreservation. Moreover, it relates the injuries to motility and fertilization capacity. Sexually mature scallops were collected from scallops farmed in La Herradura Bay, Coquimbo - Chile. The male portion of the gonad was cut in pieces (5 mm), which were after placed in a petri dish containing filtered sea water. Liberation of sperm was induced, which made it possible to obtain a spermatic solution. The sperm was equilibrated for 5 min in a crioprotective solution containing 10% ME2SO, 125 mM sucrose and 10% yolk of hen eggs diluted in sea water. Subsequently, the amples were frozen at a rate of 8,8 ºC/min. After 24 hours, the samples were firstly thawed while immersed for 20 seconds in water of 50º C, finally they were thawed while immersed in water at room temperature. The percentage of motility, fertilization of fresh oocytes and injured spermatozoa (in head, acrosome, middle piece and tail) were quantified with a light microscopy, stereoscopy and scanned electron microscopy, respectively. Both the freeze-thawing treatment and the post dilution time had significant effects on the spermatic motility. Moreover, it was observed a significant interaction between the post-dilution time and the freeze-thaw treatment (C, PRE o POST). Spermatozoa exposed to the pre-freezing treatment (PRE) remained motile for a longer while than the ones exposed to the control treatment (C). The motility in the spermatozoa in the post-thaw treatment was always much lower than in the PRE and C treatments. The morphology of the spermatozoids was affected in several ways by the freeze-thawing treatment. Some had their head deformed or swollen, others had their cell membrane folded or broken. Acrosome reaction, anomalous positions or absence of mitochondria were also observed. Broken or stiff or lineal structure loosed of tail were the most frequent injuries. The C and PRE had higher percentage of unhurt sperm (87,7 ± 3,4 % and 79,0 ± 3,0 % respectively), while the PRE samples had 14,2 ± 2,8 % of unhurt sperm. The tail was the spermatic structure that most commonly injured during the freezing-thawing process (77,0 ± 3,0 %). The percentage of sperm with head injury was 55,1 ± 7,4 % and with acrosome reaction was 28,7 ± 3,3 %. The middle piece was affected in 23,9 ± 4,1 % of the spermatozoa. The percentage of fertilization was 68,3 ± 6,6 %, 67,9 ± 4,2 % and 58,2 ± 7,3 % for C, PRE and POST respectively, which were not significantly different. The injuries were correlated with motility (at 5 and 30 min) and with fertilization success. There was a better correlation between injuries and motility than between injuries and fertilization success. The correlation between motility and fertilization was low (0,605 and 0,668 with motility at 5 min and 30 min respectively). The crioprotective solution did not have any toxic effects on the spermatozoa, on the contrary, it rather had an activating effect on motility. Moreover, the crioprotective solution provided protection during the freezing thawing process, which is a critical process in cryopreservation in which spermatozoa suffer of decreased motility. The crioprotective solution did not have a significant effect on the external morphology of the spermatozoa. Hence, again the freezing thawing process is causing the injuries. The heads destroyed of the spermatozoa would be consequence of the accumulation of ice crystals. The tails could be rigid due to alterations in the axonomic structure. Possibly the acrosome reaction could be by the bipolarity changes due to the process of freezing-thawing. As a rule, the injury in any structure of the spermatozoa is related to the cell membrane.

ROCHA, V. P. Filogenia morfológica e molecular e aspectos biogeográficos da subfamília chioninae (Bivalvia: veneridae). 2016. 121 f. Tese (Doutorado em Ciências Marinhas Tropicais) - Instituto de Ciências do Mar, Universidade Federal do Ceará, Fortaleza, 2016. / Submitted by Geovane Uchoa (geovane@ufc.br) on 2016-06-24T15:32:56Z No. of bitstreams: 1 2016_tese_vprocha.pdf: 5549832 bytes, checksum: 086e4291a3c9cb6ab828911520b7e337 (MD5) / Approved for entry into archive by Nadsa Cid (nadsa@ufc.br) on 2016-06-24T16:04:58Z (GMT) No. of bitstreams: 1 2016_tese_vprocha.pdf: 5549832 bytes, checksum: 086e4291a3c9cb6ab828911520b7e337 (MD5) / Made available in DSpace on 2016-06-24T16:04:58Z (GMT). No. of bitstreams: 1 2016_tese_vprocha.pdf: 5549832 bytes, checksum: 086e4291a3c9cb6ab828911520b7e337 (MD5) Previous issue date: 2016

Submitted by Ginamara Lima (ginaj@pucrs.br) on 2012-05-04T12:42:51Z No. of bitstreams: 1 437942.pdf: 14499337 bytes, checksum: 95040a95e0a346b6d9760a8fda191ce6 (MD5) / Made available in DSpace on 2012-05-04T12:42:51Z (GMT). No. of bitstreams: 1 437942.pdf: 14499337 bytes, checksum: 95040a95e0a346b6d9760a8fda191ce6 (MD5) / Este estudo teve como objetivo avaliar a citotoxicidade e bioatividade do cimento de ionômero de vidro modificado com carbonato de cálcio de conchas. Para os testes de toxicidade celular foi utilizando a Saccharomyces cerevisiae como organismo modelo. A indução de citotoxicidade foi avaliada por dois testes diferentes usando a S. Cerevisiae cepa selvagem wild-type FF18733: (1) a sobrevivência da célula de levedura e (2) formação de colônias petite (mutantes respiratórios). Para os testes de bioatividade foi avaliada a variação de peso das amostras e análise de deposição de hidroxiapatita pela MEV e EDS após imersão em solução simuladora de plasma sangüíneo. Os resultados dos testes de sobrevivência mostraram que as diferentes concentrações de carbonato de cálcio adicionado ao CIV (1%, 5% e 10% em peso) induziram uma ligeira perda de viabilidade celular em S. cerevisiae em relação ao controle negativo, porém não foi o suficiente para ser considerado como uma indução de toxicidade significativa. Em relação às colônias petite, não foi observada a indução de formação de mutantes respiratórios em qualquer uma das concentrações testadas, indicando que o CIV modificado não induziu estresse oxidativo em células de S. cerevisiae. Na avaliação da bioatividade, os resultados mostraram que após a imersão todos os grupos testados apresentaram uma diminuição no peso das amostras, que pode ser atribuído à dissolução do cimento de ionômero de vidro. Na quarta semana de imersão houve uma variação significativa na percentual médio de peso para todos os grupos. A análise de MEV mostrou uma ligeira deposição de hidroxiapatita sobre a superfície dos grupos com a adição de pó de conchas em comparação ao grupo controle. / This study aimed to evaluate the cytotoxicity and bioactivity of the glass ionomer cement modified with calcium carbonate shells. For testing cell toxicity was using Saccharomyces cerevisiae as a model organism. The induction of cytotoxicity was evaluated by two different tests using wild-type strain FF18733 of S. cerevisiae: (1) yeast cell survival and (2) formation of petite colonies (respiratory mutants). For tests of bioactivity was evaluated weight change of samples and analysis of hydroxyapatite deposition by SEM and EDS after immersion in simulated blood plasma. The results of survival tests showed that different concentrations of calcium carbonate added to the GIC (1%, 5% and 10% by weight) induced a slight loss of cell viability in S. cerevisiae in relation to the negative control, but it was not enough to be considered as a significant induction of toxicity. In relation to the petite colonies, there was no induction of mutants respiratory formation in any of the concentrations tested, indicating that the GIC did not induce altered oxidative stress in cells of S. cerevisiae. In the evaluation of bioactivity, the results showed that after immersion all groups tested showed a decrease in weight of the samples, which can be attributed to the dissolution of glass ionomer cement. In the fourth week of immersion there was a significant variation in the average percentage of weight for all groups. The SEM analysis showed a slight deposition of hydroxyapatite on the surface of the groups with the addition of powdered shells compared to the control group.

El presente trabajo de tesis tiene como objetivo identificar y cuantificar las diferentes alteraciones morfológicas de espermatozoides de Argopecten purpuratus, ocasionadas por el proceso de criopreservación, y relacionarlas con su motilidad y capacidad de fecundación post-descongelamiento. Se utilizaron conchas de abanico maduras obtenidas de cultivos suspendidos de la Bahía la Herradura de Guayacán, Coquimbo-Chile. Se disectó la gónada y la porción masculina se cortó en trozos para liberar los espermatozoides en AMFE. Estos, fueron incubados en una solución crioprotectora compuesta por ME2SO al 10%, 125 mM de sacarosa y 10% de vitelo del huevo de gallina. Se congeló los espermatozoides a una tasa de 8,8 ºC/min. El descongelamiento se realizó 24 horas después mediante inmersión en un recipiente con agua a 50º C durante 20 seg y luego en otro a temperatura ambiente. Posteriormente se cuantificó el número de espermatozoides móviles en un microscopio de luz; también se cuantificó la cantidad de espermatozoides lesionados en la cabeza, acrosoma, pieza media y flagelo en un microscopio electrónico de barrido. Finalmente, se determinó la capacidad fecundante de los espermatozoides inseminando ovocitos frescos. La motilidad varió significativamente entre los tratamientos (C, PRE y POST) en cuanto a porcentaje de espermatozoides móviles y tiempo de actividad. Se observó distintas alteraciones en la morfología del espermatozoide congelado-descongelado. Algunas tuvieron la cabeza deforme, hinchada o lisada y la membrana celular plegada o rota. También fue observada, reacción acrosómica y mitocondrias fuera de su posición normal o ausentes. Pero las alteraciones más comunes se encontraron en el flagelo, el cual sufrió ruptura, rigidez y pérdida de su estructura lineal. En C y PRE el porcentaje de espermatozoides ilesos fue mayor (87,7 ± 3,4 % y 79,0 ± 3,0 % respectivamente), mientras que en POST fue 14,2 ± 2,8 %. La estructura lesionada en la mayor cantidad de espermatozoides POST, fue el flagelo (77,0 ± 3,0 %); el porcentaje de espermatozoides con lesión en la cabeza fue 55,1 ± 7,4 % y los que tuvieron el acrosoma reaccionado fueron 28,7 ± 3,3 %. La pieza media fue afectada en el 23,9 ± 4,1 % de espermatozoides. Los porcentajes de fecundación fueron 68,3 ± 6,6 %, 67,9 ± 4,2 % y 58,2 ± 7,3 % para C, PRE y POST respectivamente, no encontrándose diferencias significativas entre ellos. Se correlacionó la motilidad total cuantificada a los 5 y a los 30 minutos, las lesiones ocurridas en las diferentes estructuras espermáticas y los porcentajes de fecundación; encontrándose mayor correlación entre las alteraciones y la motilidad que entre las alteraciones y los porcentajes de fecundación. La correlación entre la motilidad y fecundación fue baja (0,650 y 0,668 con la motilidad a los 5 y 30 min respectivamente). La incubación durante el tiempo de equilibrio en la solución crioprotectora utilizada, no tiene un efecto tóxico que altere la motilidad o la morfología de los espermatozoides, sino más bien, tiene un efecto activador de la motilidad. Esta propiedad de la solución crioprotectora, se suma a la de protección, que les daría a los espermatozoides durante el congelamiento-descongelamiento, que es la etapa del proceso de criopreservación durante la cual los espermatozoide son afectado negativamente.. Las cabezas destrozadas de los espermatozoides sería consecuencia de la lisis celular o del crecimiento de cristales de hielo, todo esto como consecuencia de desórdenes osmóticos. La rigidez de los flagelos podría ser por alteraciones en la estructura del axonema. Posiblemente, la reacción acrosomal sería ocasionada por los cambios de polaridad ocurridos por el congelamiento-descongelamiento. En general, las lesiones en cualquiera de las estructuras espermáticas están relacionadas finalmente con la membrana celular. / --- The present work identifies and quantifies the morphological external alterations of spermatozoa of the scallop A. purpuratus due to long-term cryopreservation. Moreover, it relates the injuries to motility and fertilization capacity. Sexually mature scallops were collected from scallops farmed in La Herradura Bay, Coquimbo - Chile. The male portion of the gonad was cut in pieces (5 mm), which were after placed in a petri dish containing filtered sea water. Liberation of sperm was induced, which made it possible to obtain a spermatic solution. The sperm was equilibrated for 5 min in a crioprotective solution containing 10% ME2SO, 125 mM sucrose and 10% yolk of hen eggs diluted in sea water. Subsequently, the amples were frozen at a rate of 8,8 ºC/min. After 24 hours, the samples were firstly thawed while immersed for 20 seconds in water of 50º C, finally they were thawed while immersed in water at room temperature. The percentage of motility, fertilization of fresh oocytes and injured spermatozoa (in head, acrosome, middle piece and tail) were quantified with a light microscopy, stereoscopy and scanned electron microscopy, respectively. Both the freeze-thawing treatment and the post dilution time had significant effects on the spermatic motility. Moreover, it was observed a significant interaction between the post-dilution time and the freeze-thaw treatment (C, PRE o POST). Spermatozoa exposed to the pre-freezing treatment (PRE) remained motile for a longer while than the ones exposed to the control treatment (C). The motility in the spermatozoa in the post-thaw treatment was always much lower than in the PRE and C treatments. The morphology of the spermatozoids was affected in several ways by the freeze-thawing treatment. Some had their head deformed or swollen, others had their cell membrane folded or broken. Acrosome reaction, anomalous positions or absence of mitochondria were also observed. Broken or stiff or lineal structure loosed of tail were the most frequent injuries. The C and PRE had higher percentage of unhurt sperm (87,7 ± 3,4 % and 79,0 ± 3,0 % respectively), while the PRE samples had 14,2 ± 2,8 % of unhurt sperm. The tail was the spermatic structure that most commonly injured during the freezing-thawing process (77,0 ± 3,0 %). The percentage of sperm with head injury was 55,1 ± 7,4 % and with acrosome reaction was 28,7 ± 3,3 %. The middle piece was affected in 23,9 ± 4,1 % of the spermatozoa. The percentage of fertilization was 68,3 ± 6,6 %, 67,9 ± 4,2 % and 58,2 ± 7,3 % for C, PRE and POST respectively, which were not significantly different. The injuries were correlated with motility (at 5 and 30 min) and with fertilization success. There was a better correlation between injuries and motility than between injuries and fertilization success. The correlation between motility and fertilization was low (0,605 and 0,668 with motility at 5 min and 30 min respectively). The crioprotective solution did not have any toxic effects on the spermatozoa, on the contrary, it rather had an activating effect on motility. Moreover, the crioprotective solution provided protection during the freezing thawing process, which is a critical process in cryopreservation in which spermatozoa suffer of decreased motility. The crioprotective solution did not have a significant effect on the external morphology of the spermatozoa. Hence, again the freezing thawing process is causing the injuries. The heads destroyed of the spermatozoa would be consequence of the accumulation of ice crystals. The tails could be rigid due to alterations in the axonomic structure. Possibly the acrosome reaction could be by the bipolarity changes due to the process of freezing-thawing. As a rule, the injury in any structure of the spermatozoa is related to the cell membrane. / Tesis

La concha de abanico (Argopecten purpuratus) es un molusco bivalvo de importancia económica para el Perú, debido a su alto valor nutritivo y su amplia aceptación en los mercados de Estados Unidos, Japón y Europa. En el Perú, no se han realizado estudios a nivel genético poblacional en esta especie, existiendo solamente estudios morfométricos, cuyas variaciones estarían ligadas a cambios en la variabilidad genética del recurso (Cisneros et al., 2008). Además, existen pocos marcadores microsatélites reportados, los cuales son los marcadores moleculares más adecuados para evaluar la diversidad genética de una especie. El objetivo del presente estudio fue detectar microsatélites trinucleótidos “CAT” del genoma de A. purpuratus para lo cual se utilizó el método de librerías genómicas enriquecidas. Se optimizó un protocolo para extracción de DNA Genómico de alta calidad a partir de la gónada masculina y utilizando como buffer de lisis TNES-urea. Asimismo, se construyó una librería genómica enriquecida utilizando esferas magnéticas lo que permitió obtener un elevado porcentaje de clones positivos (65 80%). Finalmente, se obtuvo DNA plasmídico de alta calidad cuyo secuenciamiento servirá para diseñar cebadores que permitan evaluar el polimorfismo de los microsatélites. Palabras clave: concha de abanico, Argopecten purpuratus, librerías genómicas enriquecidas, microsatélites, polimorfismo. / --- The Peruvian scallop (Argopecten purpuratus) is an economically important bivalve in Peru due to its high nutritional qualities and its wide acceptance in the markets of United States, Japan and Europe. In Peru, there have not been studies of this resource at the population genetic level and there are only morphometric studies, one of those reports that variations at this level could be linked to the genetic variability of the resource (Cisneros et al., 2008). In addition, there are only few microsatellite markers reported, this molecular markers are the best to assess the genetic diversity. Due to this reason, the aim of this study was to detected trinucleotide microsatellites with the motif "CAT" from the genome of A. purpuratus using enriched genomic libraries. High quality DNA extraction from the male gonad with TNES-urea lysis buffer was optimized. Furthermore, the enriched genomic libraries constructed using magnetic beads allowed identification of a high percentage of positive clones (65-80%). Finally, the sequencing of the high quality plasmid DNA obtained will enable primers design for the evaluation of the microsatellites polymorphism. Keywords: Peruvian scallop, Argopecten purpuratus, enriched genomic libraries, microsatellite, polymorphism. / Tesis