Orientador: Antonio Leão Castilho / Abstract: The present study was divided into two chapters that aimed to verify the spatiotemporal abundance of juveniles and adults, analyzing how the variation of the environmental factors (salinity, water temperature, rainfall, phi, organic matter) affect the distribution patterns and describe the population dynamics of Litopenaeus schmitti in Cananéia, south coast of the São Paulo state, Brazil. With a shrimp boat the shrimp and environmental factors were sampled monthly from July 2012 to May 2014, covering both the marine and estuarine environment. In the laboratory, shrimps were separated according to sex, measured on carapace length (CC) and classified for gonadal development stage. Growth and longevity were estimated from the von Bertalanffy equation and the sex ratio was tested by applying the Binomial test. A total of 1102 individuals were collected, 273 juveniles and 829 adults, with CC ranging from 16.7 to 39.6 mm in males and 16.7 to 44.9 mm in females. Adults were captured predominantly in sampling stations with marine influence, while juveniles were found in all sampling stations, especially those with estuarine influence. The presence of the majority of juvenile individuals in the estuarine environment evidences the importance of the estuary in the ontogeny and protection of the species, since this environment presents physical barriers (wide variation of salinity) for its predators. The environmental factor that most influenced the juveniles was the temperature and the... (Complete abstract click electronic access below) / Resumo: O presente estudo foi dividido em dois capítulos que tiveram como objetivos verificar a abundância espaço-temporal de juvenis e adultos, analisando como a variação dos fatores ambientais (salinidade, temperatura da água, pluviosidade, phi, matéria orgânica) afetam os padrões de distribuição da espécie, e descrever a dinâmica populacional de Litopenaeus schmitti em Cananéia, litoral sul do estado de São Paulo, Brasil. Com um barco camaroeiro os camarões e os fatores ambientais foram amostrados mensalmente de julho de 2012 a maio de 2014, abrangendo tanto o ambiente marinho quanto o estuarino. Em laboratório, os camarões foram separados quanto ao sexo, mensurados quanto ao comprimento da carapaça (CC) e classificados quanto ao estágio de desenvolvimento gonadal. O crescimento e a longevidade foram estimados a partir da equação de Von Bertalanffy e a razão sexual foi testada aplicando-se o teste Binomial. Um total de 1102 indivíduos foi coletado, sendo 273 juvenis e 829 adultos, com o CC variando de 16,7 a 39,6 mm nos machos e 16,7 a 44,9 mm nas fêmeas. Os adultos foram capturados predominantemente nas estações amostrais com influência marinha, enquanto que os juvenis foram encontrados em todas as estações amostrais, com destaque as que possuíam influência estuarina. A presença da maioria dos indivíduos juvenis no ambiente estuarino evidencia a importância do estuário na ontogenia e proteção da espécie, uma vez que este ambiente apresenta barreiras físicas (ampl... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Programa de Pós-Graduação em Biologia Celular e do Desenvolvimento, Florianópolis, 2015 / Made available in DSpace on 2015-06-02T04:08:15Z (GMT). No. of bitstreams: 1 333914.pdf: 1463583 bytes, checksum: 4b55a96be627a5ad68a55463453ece36 (MD5) Previous issue date: 2015 / A incidência de doenças virais tem implicado em significativas perdas econômicas para a carcinicultura. Nesse contexto, a busca por estratégias que aumentem a capacidade dos camarões de sobreviver a surtos virais é de grande interesse para a aquicultura, como ativar as defesas antivirais utilizando a tecnologia de RNA de interferência (RNAi). O presente trabalho avaliou o efeito do silenciamento gênico do vírus da mionecrose infecciosa (IMNV) na sobrevivência de camarões Litopenaeus vannamei e no perfil transcricional de genes associados ao seu sistema imune. Animais previamente tratados com dsRNA específico para o genoma do IMNV (dsRNA-IMNV) foram desafiados com uma dose letal desse mesmo vírus (1,02×106 cópias virais), enquanto camarões do grupo controle foram tratados com dsRNA vírus-inespecífico (gene IGSF4D de Danio rerio). O tratamento com dsRNA-IMNV conferiu uma proteção específica contra o IMNV, levando a uma sobrevivência de 90 % (30 dias), enquanto todos os animais tratados com dsRNA inespecífico morreram em 17 dias. A expressão de 46 genes, de diferentes categorias funcionais, foi quantificada por PCR quantitativa em tempo real nos diferentes grupos experimentais nos tempos 0 h, 24 h e 48 h pós-infecção. Dos genes analisados, 39 (85 %) apresentaram a expressão inalterada em todas as condições analisadas. Por outro lado, seis genes (13 %) pertencentes às categorias funcionais defesa antiviral (Lv-Ago2 e LvDcr2), defesa antimicrobiana (Litvan ALF-D e LvCrustin), reconhecimento (LvGal) e apoptose (LvIAP) foram diferencialmente expressos entre os animais que sobreviveram ou não sobreviveram à infecção por IMNV. Em conjunto, a expressão desses genes constitui uma assinatura transcricional associada a uma resposta imune eficaz contra o IMNV. De maneira interessante, o gene de coagulação (LvClot) apresentou expressão diferenciada apenas em função do tratamento com o dsRNA inespecífico (24h e 48 h pós-desafio), apresentando maiores níveis de transcritos nos animais que sucumbiram à infecção. A expressão de LvClot pode representar assim um indicativo precoce de estado mionecrótico em camarões. Com base nos resultados obtidos no presente estudo, foi possível identificar uma assinatura transcricional em hemócitos de L. vannamei associada à indução de proteção antiviral específica nos camarões, utilizando a técnica de RNAi. Assim, este é o primeiro relato da identificação de efetores do sistema imune potencialmente envolvidos na proteção e sobrevivência de camarões frente à infecção pelo IMNV.<br> / Abstract: The incidence of viral diseases has been implied in significant economic losses to the shrimp farming industry. In this context, strategies to improve the ability of shrimp to survive viral outbreaks are a major goal for aquaculture, such as the activation of shrimp antiviral defenses by using the RNA interference (RNAi) technology. The present study evaluated the effect of the RNAi-mediated gene silencing of the Infectious Myonecrosis Virus (IMNV) on the survival of Litopenaeus vannamei shrimp and on the expression profile of genes associated to its immune system. Animals previously treated with dsRNA specific to the IMNV genome (dsRNA-IMNV) were challenged with a lethal dose of the virus (1.02×106 viral copies), while shrimp from the control group were treated with a nonspecific dsRNA (dsRNA-IGSF4D from Danio rerio). The dsRNA-IMNV treatment provided a specific protection against the IMNV, reaching a survival rate of 90 % (up to 30 days), whereas all animals treated with the nonspecific dsRNA died within 17 days. The expression of 46 genes, from different functional categories, was assessed in all experimental groups at 0 h, 24 h and 48 h post-infection. From those, 36 genes (85 %) did not change their expression pattern in all analyzed conditions. On the other hand, six genes (13 %) from the functional categories antiviral defense (Lv-Ago2and LvDcr2), antimicrobial defense (Litvan ALF-D and LvCrustin), microbial recognition (LvGal) and apoptosis (LvIAP) were differentially expressed between shrimp that survived and not survived to IMNV infection. Taken together, the expression of these genes can be considered as a transcriptional signature associated with an efficient immune response against IMNV. Interestingly, the coagulation gene (LvClot) was differently expressed only in the treatment with nonspecific dsRNA (24 h and 48 h post-infection), showing higher levels of transcripts in animals that succumbed to the infection. The expression of LvClot could be an indicative of an initial state of myonecrosis in shrimps. Based on these results, it was identified a hemocyte transcriptional signature in L. vannamei associated with the induction of a specific antiviral protection in shrimp, using the RNAi approach. Therefore, this is the first report on the identification of immune effectors potentially involved in shrimp protection and survival to IMNV infection.

Orientador: Fernando José Zara / Resumo: Esta dissertação visou estudar o sistema reprodutor masculino do camarão Xiphopenaeus kroyeri com o intuito de esclarecer a maturação dos machos e incluir mais informações sobre a morfologia e reprodução dos animais da tribo Trachypeneini. No primeiro capítulo foi testada a sincronia entre a maturidade morfológica, determinada pela presença de petasma, e a maturidade fisiológica, determinada pela observação microscópica de espermatozoides e espermatóforos no vaso deferente (VD). Para isso, o sistema reprodutor de indivíduos machos juvenis e adultos foi analisado histologicamente e descrito. A partir da classe de tamanho de 12 ┤13 mm de comprimento de carapaça (CC) os indivíduos apresentam petasma, contudo a presença de espermatozoides e espermatóforos no sistema reprodutor deu-se somente a partir da classe de 14 ┤15 mm de CC. O sistema reprodutor apresenta um par de testículos lobados fusionados, onde são produzidos os espermatozoides, os quais se mostram prontos ainda neste órgão. Os espermatozoides são direcionados aos VD, os quais são divididos em região proximal (PVD), onde os espermatóforos são montados, e regiões média (MVD) e distal (DVD), onde são produzidos os principais componentes do fluido seminal. Ao final do VD encontra-se a ampola, uma glândula adjacente ao VD que produz secreção ausente de espermatozoides e espermatóforos. No segundo capítulo, foram descritas ultraestruturalmente a espermatogênese e a produção de secreção pelo VD. No testículo, as células germ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In this work, we analyzed the male reproductive system of the shrimp Xiphopenaeus kroyeri to clarify the male maturation and add more information about the morphology and reproduction of the tribe Trachypeneini. In the first chapter, we investigated the synchrony between the morphological maturity, determined by the presence of petasma, and the physiological maturity, determined by the macroscopic observation of spermatozoa and spermatophores in the vas deferens (VD). For this, the male reproductive system of juveniles and adult males was histologically analyzed and described. From the size class of 12 ┤ 13 mm of carapace length (CL) the individuals presented petasma, however the presence of spermatozoa and spermatophores was detected in the male reproductive system from the size class of 14 ┤ 15 mm of CL. The reproductive system presented a pair of fusioned lobed testes, where the spermatozoa are produced, which are fully matured in this organ. The spermatozoa are directed to the VD, which are divided in proximal region (PVD), where the spermatophores are packed, and medial (MVD) and distal (DVD) regions, where the main compounds of the seminal fluid are produced. At the end of the VD is located the ampoule, an adjacent glandule which produces secretion absent of spermatozoa and spermatophores. In the second chapter, the spermatogenesis and the seminal fluid production were ultrastructurally described. In the testis, the germ cells showed three stages of spermiogenesis, beco... (Complete abstract click electronic access below) / Mestre

Another important gene, heat shock factor (MeHSF) was also cloned using homology based PCR because it was suggested to participate in the transcriptional regulation of many essential components of ovarian maturation including vitellogenin gene and several proteins for hormones metabolism. The complete cDNA sequence of MeHSF was 2211 bp in length, which encoded a 622 amino acid protein. The translated MeHSF protein shared high similarity with those of other species, especially in the N terminal region. RT-PCR showed that MeHSF was universally expressed in most of the female tissues investigated including ovary, central nervous system, heart, gill, gut and muscle. However, its expression was not detectable in eyestalk and hepatopancreas. MeHSF was highly expressed in immature ovaries, and decreased dramatically with the progress of ovarian maturation. Since the synthesis of vitellogenin in ovary showed an opposite trend, the result suggested that MeHSF probably functioned as a transcriptional repressor to vitellogenin. Four HSFs isoforms generated from alternative splicing were obtained in immature ovaries, suggesting a possible universal role of HSF in coordinating transcription of different target genes during shrimp ovarian maturation. / As an important component of enzymatic scavenger systems, glutathione peroxidases (GPx) play important roles in maintaining the balance between reactive oxygen species (ROS) production and cellular scavenging ability. In this research, a full length GPx gene (MeGPx) which had been identified using RAP-PCR previously was cloned and characterized. MeGPx might play a pivotal role in preventing oocytes from oxidative damage and balancing ROS production. The present data on shrimp GPx provides insights on the regulation of ROS in the ovarian maturation process. / Four candidate genes possibly participating in the regulation of ovarian maturation were obtained by random sequencing the libraries, including metallothionein, two zinc finger proteins and member 4 of wingless-type MMTV integration site family (WNT4). The zinc finger protein containing a plant homeodomain, was only expressed in the eyestalk of female with immature ovaries, but not that of female with early mature and mature ovaries. The full length cDNA sequence of shrimp WNT4 gene (MeWNT4) was obtained using RACE technique. RT-PCR showed that the expression of MeWNT4 in eyestalk decreased with the maturation of shrimp ovaries. Interestingly, MeWNT4 was strongly expressed in the central nervous system and gut of both female and male shrimp. It was suggested that WNT4 could antagonize the testis determining factor (SRY), and play an essential role in suppressing the formation of testis, and at the same time, controlling of female development. Thus, the identification WNT4 from crustacean would contribute to our understanding on the sex determination mechanism. / In this thesis, two genes, heat shock protein 90 and heat shock factor, possibly playing important roles in shrimp ovarian maturation were identified and characterized. / Shrimp farming plays an important economic role in Southeast Asian countries. Yet further development of this industry is seriously restricted by the environmental deterioration and the prevalence of fetal diseases. Moreover, the failure of sexual maturation of cultured female shrimp forms a bottleneck to the further development of shrimp aquaculture. With the aim to produce shrimp without totally depending on the wild stocks, many studies have been focused on the endocrine regulation of shrimp ovarian maturation. In order to enhance our understanding on the molecular events occurred during ovarian maturation, in this research, several candidate genes are identified, and their potential roles in ovarian maturation are studied in the shrimp Metapenaeus ensis. / Since heat shock protein 90 gene is one of the essential components of steroid hormone signal cascades in vertebrates, it was cloned and isolated by homology cloning strategy. The complete cDNA sequence of shrimp Hsp90 ( MeHsp90) was 2524 by in length, which encoded a 720 amino acid polypeptide. The MeHsp90 coding region was interrupted by four introns. MeHsp90 was differentially expressed in eyestalk, ovary and hepatopancreas at different ovarian maturation stages, and consistently expressed in other tissues including heart, gill, gut, muscle and central nervous system. In vitro ovary explant assay revealed that MeHsp90 expression in immature ovary could be induced by the addition of exogenous estradio1-17beta. MeHsp90 was highly expressed in pre-vitellogenic oocytes, and its expression decreased with the progress of maturation, and finally stopped in late-vitellogenic oocytes. The co-regulation of MeHsp90 and vitellogenin by estrogen hormone suggested a possible regulatory role of Hsp90 in vitellogenin synthesis of the shrimp. / Wu, Long Tao. / "May 2008." / Adviser: Chu Ka Hou. / Source: Dissertation Abstracts International, Volume: 70-03, Section: B, page: 1409. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (p. 92-113). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Ma, Ka Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 88-103). / Abstracts in English and Chinese. / ABSTRACT --- p.i / ACKNOWLEDGEMENTS --- p.vii / CONTENTS --- p.ix / LIST OF TABLES --- p.xi / LIST OF FIGURES --- p.xii / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Molecular phylogenetics --- p.1 / Chapter 1.2 --- Phylogeny of the penaeoid shrimps --- p.2 / Chapter 1.2.1 --- Interfamilial relationships of Penaeoidea --- p.3 / Chapter 1.2.2 --- Ingergeneric relationships of Penaeidae --- p.8 / Chapter 1.2.3 --- Interspecific relationships of Penaeus s.l --- p.11 / Chapter 1.3 --- Molecular markers for decapods phylogenetics studies --- p.14 / Chapter 1.3.1 --- Mitochondrial markers --- p.14 / Chapter 1.3.2 --- Nuclear markers --- p.16 / Chapter Chapter 2 --- Molecular phylogeny of superfamily Penaeoidea / Chapter 2.1 --- Introduction --- p.19 / Chapter 2.2 --- Materials and methods --- p.21 / Chapter 2.3 --- Results --- p.28 / Chapter 2.4 --- Discussion --- p.40 / Chapter 2.5 --- Conclusions --- p.48 / Chapter Chapter 3 --- Molecular phylogeny of genus Penaeus sensu lato / Chapter 3.1 --- Introduction --- p.50 / Chapter 3.2 --- Materials and methods --- p.50 / Chapter 3.3 --- Results --- p.56 / Chapter 3.4 --- Discussion --- p.74 / Chapter 3.5 --- Conclusions --- p.84 / Chapter Chapter 4 --- General conclusions --- p.85 / References --- p.88

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Programa de Pós-Graduação em Biotecnologia, Florianópolis, 2011 / Made available in DSpace on 2012-10-26T00:21:17Z (GMT). No. of bitstreams: 1 289632.pdf: 1916144 bytes, checksum: 67af3bb6cca1c4c3c30dc049f92d0a4b (MD5) / O vírus da mionecrose infecciosa (IMNV) causa uma doença progressiva em camarões de cultivo com substanciais perdas econômicas no Brasil e na Indonésia. Métodos imunológicos simples e rápidos para a detecção do IMNV ainda não são disponíveis devido à falta de anticorpos monoclonais (AcMo) contra o vírus. Neste trabalho, dois fragmentos do gene da proteína do capsídeo do IMNV, abrangendo os aminoácidos 105-297 (IMNV105-297) e 300-527 (IMNV300-527), foram clonados e expressos em Escherichia coli. Ambas sequências nucleotídicas e aminoacídicas deduzidas de IMNV105-297 e IMNV300-527 apresentaram elevada identidade com sequências de IMNV isolados no Brasil (99%) e na Indonésia (98%). Dentre os hibridomas que foram obtidos, seis AcMo anti-rIMNV105-297 (1.1D, 1.3H, 3.3G, 3.9G, 4.6C, 5.4H) e dez AcMo anti-rIMNV300-527 (1.3G, 1.3H, 1.8C, 2.9C, 2.9E, 3.3A, 9.7F, 9.8D, 9.8H, 11.2D) foram selecionados para se avaliar suas reatividades contra a proteína do capsídeo viral presente em lisado de camarões infectados por IMNV. Nos ensaios de imunodot-blot, cinco AcMo anti-rIMNV105-297 e oito AcMo anti-rIMNV300-527 apresentaram distintas sensibilidades contra lisados de tecido muscular infectado por IMNV, bem como contra rMNV105-297 ou rIMNV300-527. Dentre esses, três AcMo anti-rIMNV105-297 (1.3H, 4.6C, 5.4H) e cinco AcMo anti-rIMNV300-527 (1.3H, 1.8C, 2.9E, 3.3A, 11.2D) demonstraram alta especificidade contra a proteína nativa do IMNV nos ensaios de Western-blot, visto que reconheceram somente uma proteína de 100 kDa - massa esperada para a proteína do capsídeo do IMNV. Nas imunohistoquímicas, dois AcMo anti-rIMNV105-297 (1.3H, 4.6C) e quatro AcMo anti-rIMNV300-527 (1.8C, 2.9E, 3.3A, 11.2D) ligaram-se a inclusões virais presentes em fibrose muscular e em áreas de necrose coagulativa. Concluindo, esses seis AcMo foram sensíveis e específicos em todos os imunoensaios realizados e poderão ser utilizados em testes de imunodiagnóstico de rotina para prevenção e controle da disseminação do IMNV. / Infectious myonecrosis virus (IMNV) has been causing a progressive disease in farm-reared shrimp with substantial economical loses in northeastern Brazil and Indonesia. Simple and rapid immunological methods for IMNV detection are not yet available due to the lack of monoclonal antibodies (MAbs) against the virus. In this report, two fragments of the IMNV major capsid protein gene, comprising amino acids 105-297 (IMNV105-297) and 300-527 (IMNV300-527), were cloned and expressed in Escherichia coli. Both nucleotide and deduced amino acid sequences of IMNV105-297 and IMNV300-527 displayed high identity to IMNV isolated from Brazil (99%) and Indonesia (98%). Among several clones secreting antibodies against the IMNV recombinant proteins, six MAbs anti-rIMNV105-297 (1.1D, 1.3H, 3.3G, 3.9G, 4.6C, 5.4H) and ten MAbs anti-rIMNV300-527 (1.3G, 1.3H, 1.8C, 2.9C, 2.9E, 3.3A, 9.7F, 9.8D, 9.8H, 11.2D) were selected to evaluate their reactivity against the native IMNV capsid protein from IMNV-infected shrimp. In immunodot-blot, five MAbs anti-rIMNV105-297 and eight MAbs anti-rIMNV300-527 presented distinct levels of sensitivities against IMNV-infected muscle tissue homogenate and against rIMNV105-297 or rIMNV300-527. From those, three MAbs anti-IMNV105-297 (1.3H, 4.6C, 5.4H) and five MAbs anti-IMNV300-527 (1.3H, 1.8C, 2.9E, 3.3A, 11.2D) demonstrated high specificity against the native IMNV protein in Western-blot, since they only recognized a 100 kDa protein - the predicted mass of IMNV capsid protein. In immunohistochemistry, two MAbs anti-rIMNV105-297 (1.3H, 4.6C) and four MAbs anti-rIMNV300-527 (1.8C, 2.9E, 3.3A, 11.2D) bound to viral inclusions present in muscle fibrosis and coagulative necrosis areas. In conclusion, these six MAbs were sensitive and specific in all immunoassays performed herein, and can be used in routine immunodiagnostic tests to prevent and control IMNV dissemination.

Made available in DSpace on 2015-11-18T13:19:30Z (GMT). No. of bitstreams: 2 rafael_piergiorge_ioc_mest_2013.pdf: 1065627 bytes, checksum: 76cbdb2a4d850d707ebf2b5a419be9ee (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015-04-14 / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil / Xiphopenaeus kroyeri (Heller, 1862) é uma das espécies brasileiras mais importantes comercialmente, sendo explotada em toda a costa, principalmente nas regiões Sul e Sudeste. A partir da década de 60 sua explotação apresentou um grande crescimento, com oscilações até a década de 80. No entanto, após a década de 80 ocorreu um grande declínio na captura e, no período de 1990-1994, estão registrados os mais baixos valores das últimas décadas. Desde então o recurso encontra-se no limite de sustentabilidade. Em 2006, foram identificadas duas espécies crípticas em Xiphopenaeus no Atlântico. Entretanto, os órgãos responsáveis desconsideram essa informação, o que provavelmente acarretará consequências graves para uma ou ambas as espécies. O presente trabalho teve como objetivo: (1) ajustar as condições de amplificação de loci EPIC e caracterizar populações das duas espécies; (2) realizar análise populacional e demográfica histórica de Xiphopenaeus sp. I, em dez pontos ao longo da costa, através de polimorfismos do gene COI; e (3) ajustar as condições de amplificação de loci microssatélites e analisar quatro populações do Brasil. Apesar de resultarem em amplificações, os marcadores EPIC foram monofórmicos nas duas espécies. Em relação ao marcador COI, oito novos haplótipos foram observados para Xiphopenaeus sp. I e cinco para Xiphopenaeus sp. II, totalizando quinze e oito haplótipos descritos, respectivamente. Apesar da identificação de novos haplótipos, o sistema de identificação molecular descrito para a identificação das espécies, continua válido As análises de \0444 apontaram que a população de Caracas (Venezuela) é geneticamente diferenciada das localidades brasileiras, enquanto que, no Brasil, Poças é diferente de Ubatuba, Santos e Cananéia. Por meio de análises de variância, confirmou-se a existência de uma separação pela ressurgência próxima de Cabo Frio. Foi verificado também que Xiphopenaeus sp. I sofreu expansão há 99,57 mil anos, provavelmente na última glaciação. Com microssatélites, apenas as análises de variância resultaram em valores significativos. Ao fim do estudo, os dados obtidos foram agrupados com informações prévias e foi construída uma distribuição consensual para a espécie. Todos esses resultados trazem importantes contribuições para o manejo de Xiphopenaeus spp, pois, além da existência de duas espécies, Xiphopenaeus sp. I é constituída por pelo menos oito estoques geneticamente diferenciados. Este é o primeiro trabalho a realizar uma amostragem tão ampla em um estudo populacional da espécie. Além disso, os dados produzidos servirão como parâmetro de comparação para outros estudos de organismos marinhos da região / Xiphopenaeus kroyeri (Heller, 1862), is one of Brazil's most important c ommercial species being exploited across the coast mostly in the South and Southeast. From the 60's its exploitation showed large growth, with fluctuations up to the 80, but after the 80's there was a huge decline in the catch and in the period 1990-1994, are recorded the lowest values of decades. Since then t he resource is within the limits of sustainability. In 2006 were identified two cryptic species in Xiphopenaeus in the Atlantic. However, the agencies responsible disrega rd this information, which probably will result in consequences for one or bot h species. This study focus to: (1) optimize the amplification conditions of EPIC loci and characterize populations of the two species, (2) perform analysis of historical dem ographic and population Xiphopenaeus sp. I, in ten points along the coast, through COI gene polymorphisms, and (3) standardize the amplification conditions of microsatellite loci and analyze four populations of Brazil. Despite the amplifications, EPIC markers were monomorphic in both species. In COI marker, eight new haplotypes w ere observed for Xiphopenaeus sp. I and five Xiphopenaeus sp. II, totaling three eight haplotypes. Even with the identification of new haplotypes, the identificatio n system described for molecular identification of species remains valid. The Φ analysis showed that Venezuela is differentiated from Brazilian localities, while in Brazil Poças is different from Ubatuba, Santos and Cananéia. Through variance analysis was confirmed the existence of a separation next of the upwelling in Cabo Frio. We a lso noticed that Xiphopenaeus sp. I expanded on 99,570 years ago, probably in the las t glaciation. Using microsatellites markers, the variance analysis was the only with si gnificants values. At the end of this study, the data were clustered with previous i nformations and the consensus distribution was constructed for the species. All t hese results supply contributions to management of Xiphopenaeus spp., because beyond the existence of two species, Xiphopenaeus sp. I consists of at least eight genetically diffe rent stocks. This is the first work to perform as wide a sampling in a study population of the species. In addition, the data produced will serve as a point o f reference for other studies of marine organisms in the region.

Xiphopenaeus kroyeri (Heller, 1862) has a wide marine distribution, occurring from North Carolina (EUA) to Rio Grande do Sul (Brazil), being the only Penaeidae found in the Western Atlantic. The species inhabits sandy or muddy ground in shallow waters. It s an important fishery resource to communities that live on the coast. The population structure analysis of sea-bob shrimp from São Francisco River mouth, Alagoas-Brazil, was the main objective of this study, and specifically the growth rate of the species. Along the research 4267 specimens of Xiphopenaeus kroyeri were collected, from which 47.93% males and 52.07% females, proportion differing from expected (1:1). The total length for males varied from 28.27 to 108.44mm (77.16 ± 0.27) with width of 80.17mm. The female reached higher length, varying from 11.20 to 125.51mm (82.03 ± 0.33) with width of 114.31mm. The average being significantly different (p> 0.0001). The weigh/length rate has showed exponential tendency, with allometric growth pattern. Moreover, the growth curves plotted in this study show that, after 24 months, the individuals from São Francisco River mouth had not reached the total length expected to the species yet, registering inferior values compared to other regions. Recruitment peaks differ from the ones established to the region, being the main peak in August/10 and the secondary in January/10, therefore the legal protection period to the species requires a review. Data evaluated in this study suggest the importance of further research, more specifically to evaluate the sustainability of the Xiphopenaeus kroyeri population, as well as to verify the efficiency of the laws applied to juvenile protection purposes. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O camarão Xiphopenaeus kroyeri (Heller, 1862) apresenta distribuição geográfica extensa, ocorrendo da Carolina do Norte (EUA) ao estado do Rio Grande do Sul e é a única espécie do gênero do Atlântico Ocidental. Habita águas costeiras rasas com fundo de areia e lama e é um recurso pesqueiro extremamente importante para as populações em sua área de ocorrência. O objetivo deste estudo foi analisar a estrutura populacional do camarão sete-barbas na região da foz do rio São Francisco Alagoas, assim como o crescimento dos indivíduos da espécie em questão. Foram coletados 4.267 exemplares, dos quais 47,93 % eram machos e 52,07 % fêmeas, diferindo do esperado para a população (1:1). O comprimento total dos machos variou de 28,27 a 108,44 mm (77,16 ± 0,27) com amplitude de 80,17 mm. As fêmeas atingiram comprimentos maiores variando de 11,20 a 125,51 mm (82,03 ± 0,33) com amplitude de 114,31 mm. As médias foram significativamente diferentes (p> 0,0001). A relação peso/comprimento para Xiphopenaeus kroyeri apresentou tendência exponencial, com padrão de crescimento alométrico. No entanto, as curvas de crescimento plotadas neste estudo mostram que, ao fim de 24 meses, exemplares da espécie na região da foz do Rio São Francisco ainda não alcançaram o comprimento máximo teórico esperado, registrando valores inferiores aos de outras regiões. Com relação ao recrutamento, os períodos diferem daqueles que norteiam a época de defeso da espécie no Estado, sendo o pico principal registrado em agosto/10 e o secundário em janeiro/10. Os dados registrados sugerem a importância de estudos mais específicos para avaliar os níveis de sustentabilidade da população de Xiphopenaeus kroyeri assim como a necessidade de verificar a eficiência do defeso na proteção dos juvenis.

Tang Chi-Yan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 89-112). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.vi / Table of contents --- p.viii / List of Tables --- p.xi / List of Figures --- p.xii / List of Abbreviations --- p.xiv / Chapter Chapter 1. --- General introduction --- p.1 / Chapter Chapter 2. --- Literature review --- p.4 / Chapter 2.1 --- History and prevalence of food allergy --- p.4 / Chapter 2.2 --- Mechanism and clinical symptoms of food allergy --- p.6 / Chapter 2.3 --- Tropomyosin as a major shellfish allergen --- p.13 / Chapter 2.4 --- Use of animal model in the studies of food allergy --- p.22 / Chapter 2.5 --- Future approaches for treatment of food allergy --- p.27 / Chapter Chapter 3. --- Cloning and expression of recombinant tropomyosin --- p.30 / Chapter 3.1 --- Introduction --- p.30 / Chapter 3.2 --- Materials and Methods --- p.31 / Chapter 3.2.1 --- Design of PCR primers for amplification of tropomyosin gene --- p.31 / Chapter 3.2.2 --- Cloning of PCR-amplified cDNA into vector --- p.32 / Chapter 3.2.3 --- Transformation of competent E. coli Ml5 cells --- p.34 / Chapter 3.2.4 --- Confirmation of DNA sequence of the cloned vector --- p.34 / Chapter 3.2.5 --- Induction of the recombinant protein --- p.35 / Chapter 3.2.6 --- Purification and storage of the recombinant protein under native condition --- p.36 / Chapter 3.2.7 --- Concentration measurement and storage of the recombinant protein --- p.37 / Chapter 3.2.8 --- Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.38 / Chapter 3.2.9 --- Regeneration of the Ni-NTA column --- p.40 / Chapter 3.3 --- Results and discussion --- p.42 / Chapter 3.3.1 --- DNA sequence of the cloned vector --- p.42 / Chapter 3.3.2 --- Expression of the recombinant protein --- p.42 / Chapter 3.3.3 --- Sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE) --- p.43 / Chapter Chapter 4. --- Induction of hypersensitive response to shrimp tropomyosin in mice --- p.47 / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Materials and methods --- p.52 / Chapter 4.2.1 --- Mice and reagents --- p.52 / Chapter 4.2.2 --- Animal sensitization and challenge --- p.53 / Chapter 4.2.3 --- Morphological and behavioral changes --- p.54 / Chapter 4.2.4 --- Tropomyosin-specific IgE level --- p.55 / Chapter 4.2.5 --- Passive cutaneous anaphylaxis (PCA) reaction --- p.56 / Chapter 4.2.6 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.56 / Chapter 4.2.7 --- Cytokine profiles of splenoctyes --- p.58 / Chapter 4.2.8 --- Histological examination of small intestine --- p.59 / Chapter 4.2.9 --- Statistical analysis --- p.59 / Chapter 4.3 --- Results --- p.63 / Chapter 4.3.1 --- Morphological and behavioral changes after challenge --- p.63 / Chapter 4.3.2 --- Tropomyosin-specific IgE level --- p.63 / Chapter 4.3.3 --- Passive cutaneous anaphylaxis (PCA) --- p.64 / Chapter 4.3.4 --- Tropomyosin-specific cellular proliferation level of splenocytes --- p.68 / Chapter 4.3.5 --- Cytokine profiles of splenocytes --- p.70 / Chapter 4.3.6 --- Histology of small intestines --- p.76 / Chapter 4.4 --- Discussion --- p.79 / Chapter Chapter 5. --- General conclusion --- p.88 / References --- p.89

Thesis (Ph. D.)--University of Hong Kong, 1992.