O camarão corrupto Callichirus seilacheri foi erigido por Bott (1955) através de uma breve e incompleta descrição carente da maioria das ilustrações sobre as principais estruturas corporais desta espécie. Callichirus seilacheri é considerada, atualmente, como o único representante do gênero Callichirus na costa do Pacífico Leste. Sua distribuição geográfica abrange um amplo gradiente latitudinal de aproximadamente 50º graus (~13º N - 37º S) desde a praia Los Blancos (localidade-tipo), El Salvador até Lenga na costa de Chile. O estado taxonômico da grande população de C. seilacheri tem sido questionado no passado em reiteradas oportunidades. Este questionamento tem relação com a falta de estudos taxonômicos associados com estas populações e com o fato que alguns antecedentes moleculares têm demonstrado que as populações de Callichirus da costa do Pacífico de Nicarágua e México são diferentes. Considerando estes antecedentes, o primero objetivo do presente estudo foi avaliar o estado taxonômico de C. seilacheri na costa do Pacífico Leste da América Central. Para isto, foi coletado material de C. seilacheri em diferentes localidades da costa do Pacifico de América Central, incluindo a localidade-tipo desta espécie na costa de El Salvador. Conforme as observações morfológicas do material coletado, os caracteres diagnósticos de C. seilacheri incluem entre outros aspectos: (i) pedúnculos oculares com uma alongada projeção pré-córneal, cujas pontas divergem lateralmente; (ii) segmentos terminais do pedúnculo antênular e antenal com as bases paralelas; e (iii) pereiópodo maior do primeiro par de pereiópodos dos machos adultos com o mero armado de um lóbulo retangular na região ventral anterior e ísquio curvado para baixo com um gancho na margem ventral. Uma das descobertas mais relevantes foi que todas as fêmeas de C. seilacheri têm ambos gonóporos masculino e feminino, o qual representa o primeiro registro de intersexualidade no gênero Callichirus. O segundo objetivo desta tese foi avaliar se a espécie da costa chilena Callichirus garthi (Retamal, 1975), considerada como uma sinonímia-junior de C. seilacheri da costa do Pacífico da América Central, é uma espécie válida. Para isto, comparou-se o material-tipo de C. garthi e outros lotes desta espécie coletados ao longo da costa de Chile (~18º e 37º S) com os espécimes de C. seilacheri coletados na localidade-tipo (Playa Los Blancos, El Salvador) e em outras localidades da América Central. A comparação morfológica revelou importantes diferenças morfológicas entre os tipos morfológicos identificados em ambas espécies. Entre as principais diferenças e possível mencionar que C. garthi possui as pontas dos pedúnculos oculares paralelas, enquanto que em C. seilacheri são divergentes formando um angulo de 60-70°. Os machos adultos de ambas espécies compartilham quelípodos desiguais na fase adulta, mas somente, C. seilacheri tem um gancho sobre a margem ventral do ísquio e um lóbulo meral retangular. A analise morfológica do material biológico coletado em ambas regiões permite concluir que os espécimes coletados ao longo da costa do Chile pertencem à C. garthi, enquanto que os exemplares coletados na costa do Pacífico da Costa Rica pertencem à espécie C. seilacheri. Conforme a essas informações, C. garthi é uma espécie válida. Portanto, sugere-se a separação da grande população de C. seilacheri em duas diferentes populações: (i) uma pertencente ao C. seilacheri a qual ocorre desde a costa do El Salvador ate Costa Rica (~19º N 08º N), (ii) outra pertencente ao C. garthi a qual ocorre entre os 18º S e 37º S da costa chilena. A hipótese do centro abundância (HCA) é baseada na suposição de que as retricções fisiológicas limitam o desenvolvimento das populações nas bordas do seu intervalo de distribuição. Com o intuito de testar a validez desta hipótese, espécimes da espécie modelo Callichirus garthi foram coletados em sete populações distribuídas ao longo da costa chilena. Em cada população foram avaliados a densidade junto com uma serie de atributos populacionais de C. garthi (e.g. tamanho máximo do corpo, razão sexual, etc). Os principais resultados revelaram que C. garthi foi significativamente menos abundante nas populações locais, perto do centro e aumentou para as bordas norte e sul da sua distribuição na costa do Chile. No extremo sul e norte da distribuição, a estrutura da população foi composta por indivíduos de todos os tamanhos, em contraste com às localidades do centro, onde os indivíduos estavam concentrados em uma faixa de tamanho mais estreita. Surpreendentemente, nas bordas (norte e sul), à razão entre os sexos foi estremamente equilibrada. A abundância local foi levemente correlacionada com a distância entre cada localidade e o rio mais próximo e foi fortemente correlacionada com a estrutura da população. Aparentemente, a proximidade de um rio oferece as condições ideais para que C. garthi forme populações densas e equilibradas, o qual seria um reflexo do sucesso a nível individual que esta espécie atinge em direção às bordas. O modelo de distribuição de abundância de C. garthi pode ser considerado como uma exceção nas populações naturais devido que na maioria dos estudos prévios nesta matéria o modelo usualmente reportado é o modelo normal de abundância para um dos extremos. As generalizações acerca do padrão reprodutivo dos invertebrados bentônicos costeiros propõem que as espécies tropicais tenham reprodução do tipo continua, como reflexo das baixas oscilações térmicas ao longo de todo o ano (hipótese de Orton). Com o propósito de testar esta suposição em um ambiente tropical da América, foram realizados durante um ano amostragens mensais em uma população de C. seilacheri da costa do Pacífico de Costa Rica (América Central). Os resultados revelaram que C. seilacheri tem uma marcada sazonalidade reprodutiva, a qual contradiz os padrões esperados em regiões tropicais. As fêmeas ovígeras estiveram presentes desde maio até novembro, o que está estatisticamente relacionado com a diminuição da salinidade durante a estação chuvosa. A temperatura não teve influência sobre a periodicidade reprodutiva. Aparentemente, à reprodução em C. seilacheri está fortemente relacionada com a variabilidade climática da costa do Pacífico na Costa Rica, sendo desencadeada por oscilações na salinidade. Uma comparação entre C. garthi da costa chilena e C. seilacheri da costa do Pacífico da América Central revelou importantes diferenças na produção de ovos. Callichirus seilacheri produz uma media de quatro menos ovos e menores que C. garthi da costa do Chile. A variabilidade nas características reprodutivas (produção e tamanho dos ovos) e de tamanho corporal dos indivíduos observada nestas duas espécies de camarões corruptos sugerem a imposição de importantes restrições fisiológicas dadas principalmente por a temperatura em ambas às regiões da América. Os aspectos examinados no presente estudo constituem novos antecedentes sobre a taxonômia, morfologia e ecología em um dos gêneros da família Callianassidae mais representativos nas praias de areia da América. Cada um destas informações pode ser considerado de especial relevância para a conservação das populações de C. garthi e C. seilacheri no Pacífico Leste. Especialmente, porque ambas espécies são frequentemente recolhidas como isca para a pesca comercial e esportiva em algumas pequenas vilas costeiras. / The ghost shrimp Callichirus seilacheri was erected by Bott (1955) through a brief and poorly illustrated description. This species is considered as the only one current representative of genus Callichirus along the eastern Pacific. Their distribution encompasses approximately 50 degree of latitude, from Los Blancos, El Salvador (13º N) to Lenga, Chile (37º S). The identity of the large population of C. seilacheri along the Pacific coast of Americas has been questioned by different researchers whom have indicated the existence of more than one species of Callichirus for the Pacific coast of Mexico and the Pacific coast of Costa Rica and Nicaragua. Considering this information, the first focus of this thesis was realize a redescription of C. seilacheri based on specimens collected in different localities from Central America, including the type-locality of this species. According to the observations, the diagnostic features of C. seilacheri include (i) the eyestalks with elongated precorneal projection with tips diverging laterally; (ii) terminal segments of antennular peduncle and antennal with the parallel base and (iii) adult male with larger cheliped with rough rectangular lobe over anterior ventral margin of merus, ischium curved downward with a hook on ventral margin. All females analyzed herein have both male and female gonopores, thus firstly recorded the presence of intersex within the genus Callichirus. The second objective of this thesis was test whether Callichirus garthi (Retamal, 1975) which is considered as a junior synonymy of C. seilacheri constitute a valid species for the eastern Pacific. For this, a full comparison between specimens of the genus Callichirus collected along the Pacific coast of Central America and Chilean coast, including the type-locality of C. seilacheri and C. garthi was realized. The comparison revealed important morphological differences between both species. For example, in C. garthi the tips of the eyestalk are parallel while diverge distally forming an about 60-70° angle in C. seilacheri. Adult males of both species share unequal chelipeds; however, only C. seilacheri has a hook on ventral margin of ischium and its merus is armed with a rectangular lobe over anterior ventral margin. Additional material collected along the Chilean coast and from different localities from the Pacific coast of Costa Rica allowed confirms the taxonomic status of both species in the Eastern Pacific. All specimens collected along the Chilean coast belong to C. garthi while those of Costa Rica fit to form of C. seilacheri from El Salvador. According to the results, C. garthi is a valid species. Therefore, it is postulated the separation of large population of C. seilacheri into two different populations: (i) one belonging to C. seilacheri which distributes from Mexico to Costa Rica (~19º 08º N), (ii) other assigned to C. garthi which encompasses, at least, between 18º S 37º S of the Chilean coast. The abundant-centre hypothesis (ACH) is based on the assumption that physiological constraints limit populations at the edges of their distributional range. In the third chapter is examined the applicability of the ACH in a marine system by testing whether physiological predictions are reflected in large-scale variations of life-history traits. In order to achieve this information, specimens of Callichirus garthi were collected in the intertidal zone at seven sampling sites. In each site we evaluated (i) local abundance, (ii) maximum body size, (iii) sex-ratio, (iv) length-size frequency distribution, and (v) size at maturity. The main outcomes revealed that C. garthi is a species of low occurrence which was present in seven of all visited beaches (N = 61). Callichirus garthi was clearly less abundant within local populations near the centre and increased towards the northern and southern edges of its distribution. This pattern of abundance was significantly fitted by the abundant edge model. At the southern end and northern of the distribution, population structure was composed by individuals of all sizes in contrast with localities from the centre where individuals were concentrated in a narrow size range. Surprisingly at the edges (north and south), the sex ratio was extremely balanced. Physical variables (SST, grain size) had not influences on variation of life-history traits; however, the local abundance was slightly correlated with the distance between each locality and the closest river and was strongly correlated with population structure. Overall, the observed pattern was intimately linked with input of nutrients from the closest rivers. The findings allow separate the great population of C. garthi along the Chilean coast in two kinds: (A) stable populations (at the border of distribution), and (B) unstable populations (at the centre of distribution). In both cases, the population demography is controlled by dense-dependent factors. The results underline the importance of incorporating ecological, physiological and life-history studies in future tests of the ACH. Generalizations about breeding patterns of near-shore benthic marine invertebrates have proposed that tropical species have continuous reproduction as a reflection of constant temperature throughout the year (Ortons hypothesis). Based on this assumption, it was analysed the reproductive periodicity and egg production in the intertidal ghost shrimp Callichirus seilacheri from the Central Pacific coast of Costa Rica. In order to obtain such information, a total of 449 individuals were collected monthly from December 2011 to November 2012. The results showed that females with ovaries filled with mature oocytes were present throughout the whole year. However, the presence of ovigerous females was restricted to the period between May and November which fits to a seasonal breeding cycle. Apparition of egg-bearing females was statistically correlated with the decrease of salinity detected during rainy season, but not with temperature. Compared to C. garthi from temperate waters, C. seilacheri produced on the average four times less eggs and smaller. Variability in reproductive features and body size of these species seem to reflect physiological constraints imposed by temperature in cold and warm regions. The aspects examined in each chapter of this thesis constitute new antecedents about morphology, taxonomy, and ecology of one of the most representative genus of the family Callianassidae from American coast. Each one of these informations may be considered of especial importance for the management and conservation of the Callichiruss populations from the eastern Pacific. Especially, because both species herein analyzed are frequently collected as bait for commercial and recreational fishing in some small villages along Pacific coast of America.

Synalpheus shrimp species of the gambarelloides group are the only marine organisms displaying the highest level of social functioning, eusociality. Their social hierarchies are equally complex compared to the reproductive abnormalities that have been recently discovered. For instance, snapping shrimp of the genus Synalpheus were thought to be gonochoric, i.e. developing as independent sexes, until scanning electron microscopy studies revealed intersexed gonopores in several species. This project analyzed both the species composition, and accompanying reproductive structures, of Synalpheus spp. (Caridea: Alpheidae) comprised of densely aggregating communal and pair-living colonies in the Florida Keys, Florida. Colonies of pair-living and communal Synalpheus spp. were observed from hosts Spheciospongia vesparium and Spongia sp. from hard bottom assemblages of the Florida Reef Tract in order to assess differing population structures. Comparisons were made of the measures of overall and relative abundance, frequency by species and sex category, and variation in growth by species and sex category, for each individual colony. We then used scanning electron microscopy (SEM) images to determine the secondary sexual characteristics of three species (Synalpheus brooksi, S. herricki, and S. cf. herricki) which range in social behavior. Species were widely consistent in both host choice and distribution, across all sampling areas. The abundance of communal species Synalpheus brooksi was much greater than expected at all sites, in comparison to previously published work. While Synalpheus longicarpus was reported at higher frequencies in prior studies, our results yielded a much lower frequency of this species, often found in pairs, rather than dense aggregations. Average sizes of ovigerous and non-ovigerous individuals in dense colonies of Synalpheus brooksi and S. pectiniger did not differ significantly. However, total body length of individuals differed within species groups, specifically related to sex and presence of ova. Within colonies of S. brooksi, mid-development, or ‘transitional’ individuals, were discovered in nearly all populations. However, the reproductive and social function of these individuals displaying mixed sexual characteristics could not be determined from this study. Individuals of S. brooksi displaying ‘transitional’ external morphology, i.e. masculine abdominal pleura paired with clutches of eggs, displayed higher incidences of intersex gonopores per colony than did conspecific non-ovigerous and ovigerous individuals. These results suggest that colonies of S. brooksi may be comprised of a subset of helpers, or individuals undergoing a transitional sexual development phase, similar to prior published findings of intersexed helpers among eusocial colonies (Toth and Bauer 2007). In comparison to S. brooksi colonies, nearly all colonies of S. herricki and S. cf. herricki were composed of intersex individuals. In conjunction with previous instances documented in eusocial Synalpheus paraneptunus groups, the data provide substantial evidence of intersexing at all levels of social organization in Synalpheus spp. (pair living, communal, and eusocial). These findings nonetheless provide a clearer picture of how social structure and life history influence adaptation of a particular reproductive strategy. Quantifying features of Spheciospongia vesparium populations and comparing results to neighboring hosts, such as Spongia sp., provided evidence for potential influences of host choice, and variation in growth and reproductive capacity temporally and spatially. These observations of species’ growth patterns and abundances contribute greatly to our understanding of life history of Synalpheus spp., and, furthermore, adaptation of social organization.

[pt] Neste estudo, foram determinadas as concentrações de Cu, Fe, Zn, Cd, Pb e Ni no tecido muscular e fígado de três espécies de peixe (Mugil liza – tainha, iliófago; Micropogonias furnieri – corvina, carnívoro e Trichiurus lepturus – peixe espada, piscívoro), no tecido muscular de uma espécie de camarão (Farfantepenaeus brasiliensis – camarão rosa, onívoro) e também, no tecido muscular, vísceras e ovos de uma espécie de siri (Callinectes spp. – siri azul, onívoro) coletados na Baía da Ilha Grande. As concentrações dos elementos foram determinadas por espectrometria de massa com fonte de plasma indutivamente acoplado (ICP-MS). A metodologia analítica apresentou ótima exatidão e precisão quando comparada com amostras certificadas (DORM-3 e TORT-2). As concentrações de Cu, Zn, Cd, Ni e Pb apresentaram-se abaixo dos limites estabelecidos pela legislação brasileira e americana para a ingestão humana de pescado. As maiores concentrações de Cu, Fe, Zn, Cd, Ni e Pb foram observadas nos crustáceos. Nos peixes, as maiores concentrações dos elementos analisados foram observadas no fígado, e no siri, nas vísceras. O sexo dos indivíduos não influenciou no acúmulo dos elementos para os peixes, porém influenciou na acumulação de Cu, Fe, Zn, Cd e Ni no siri. Nos peixes, a maioria dos elementos não apresentou correlação com o comprimento dos indivíduos, com exceção do Pb, que apresentou correlação negativa na tainha e na corvina, e do cádmio, que apresentou correlação negativa na corvina. Nos crustáceos, alguns elementos também apresentaram correlação negativa com estes parâmetros, indicando que, tanto nos peixes quanto nos crustáceos, estes elementos não foram acumulados ao longo do crescimento. As concentrações encontradas foram diferentes nas estações seca e chuvosa. / [en] In this study the concentrations of Cu, Fe, Zn, Cd, Pb and Ni were determined in muscle tissue and liver of three fish species (Mugil liza – mullet, illiophagus; Micropogonias furnieri – croaker, carnivorous, and Trichiurus lepturus – cutlassfish, piscivore), in the muscle tissue of a shrimp species (Farfantepenaeus brasiliensis – pink shrimp, omnivore) and in muscle tissue, entrails and eggs of a soft crab (Callinectes spp. – siri azul, omnivore) sampled from the Ilha Grande Bay. The concentrations of the elements were determined by inductively coupled plasma mass spectrometry (ICP-MS). The methodology showed good accuracy and precision when tested with certified reference materials (DORM-3 and TORT-2). The concentrations of Cu, Zn, Cd, Ni and Pb were below the limits established by the Brazilian and American legislation for human ingestion of fish products. The highest concentrations of Cu, Fe, Zn, Pb, Cd and Ni were found in the crustacean samples. In fish, the highest concentrations were observed in the liver and, in the soft crab samples, in the entrails. Sex did not influence element accumulation in fish, but influenced the accumulation of Cu, Fe, Zn, Cd and Ni in the soft crab. In fish, most elements did not show any correlation to length and weight, with the exception of Pb, which showed negative correlations in mullet and croaker and Cd which showed negative correlations in croaker. In crustaceans, some elements also presented negative correlations to these parameters, indicating that both in fish and crustaceans these elements are not accumulated during organism growth. The concentrations of the elements analyzed were statistically different in the dry and rainy seasons.

The purpose of this research was to develop and standardize a novel feeding bioassay with Macrobrachium rosenbergii for use in the laboratory and allowing it to be easily deployed under field conditions. Standardization of the test aimed to minimize feeding rate variations and to ensure that subsequent statistical analyses have sufficient power to consistently detect changes in feeding rates. These were accomplished through the development of a post-exposure feeding toxicity test under laboratory, microcosm and in situ/field conditions. This procedure was proven to be repeatable and economical. M. rosenbergii as test animals were available in terms of quantity and uniformity in sizes. The standard guidelines and procedures for M. rosenbergii bioassay developed from this study include the size of test animals (9-10 mm), density in exposure containers (10 animals in 500 mL of medium in the laboratory, 10 animals in field chambers with 98.6 mL volume), exposure time (24 hours), feeding period for post-exposure feeding (4 hours) and number of replicates for the feeding test (10 replicates for individual measurements). The tiered approach used in the preliminary risk assessment of pesticide using TOXSWA was capable of screening the risk level of pesticide in the study area, identifying profenofos and dimethoate as test chemicals for the lethal and sub-lethal experiments. This model was beneficial in the preliminary risk assessment of pesticides in the tropics, since it was not necessary to set up laboratory work. This method could also provide preliminary data to support the environmental planner and decision/policy maker. This is an alternative way to develop a cost efficient model to inform and warn the risk of pesticide use. The effects of pH, temperature and hardness on control post-exposure feeding rates of M. rosenbergii were assessed and indicated that M. rosenbergii was very sensitive to acidic and basic conditions. The use of post-exposure feeding inhibition as the endpoint under laboratory conditions revealed that prawns were sensitive to pesticides (chlorpyrifos, dimethoate and profenofos) and a heavy metal (zinc). Post-exposure feeding rate inhibition could be used as a sublethal endpoint as the EC50 values obtained for chlorpyrifos and zinc were lower than their lethal levels. Mortality of prawn was also another endpoint used to define the toxicity of pesticides such as carbendazim, in which mortality occurred during exposure, but post-exposure feeding rate of the surviving animals did not decrease. The microcosm experiments were able to link the laboratory toxicity tests and the effects observed in the field. Microcosm studies provided another dimension to studies looking at pesticide effects on aquatic systems. In this research, carbendazim affected feeding and survival rates in the microcosm set-up but in the laboratory only mortality showed a significant difference (P < 0.05). In situ bioassays were able to show the effects of pesticides on post-exposure feeding rates using the methods developed. Post-exposure feeding rates were significantly lower than control in farms using pesticides while in uncontaminated sites (pesticide-free), the post-exposure feeding rates did not decrease. However, mortality was observed even in the uncontaminated sites which could be attributed to other factors such as low dissolved oxygen and presence of some other unidentified chemical substances. The degree of mortality and the effect on feeding rates depends not only on the type and concentration of the known pesticide but also on water quality parameters. The basic methods developed for in situ bioassay from this research is a simple, easy and fast way to determine the effect of pesticides because the results can be seen in the field. The procedures developed and results obtained from this study can be used as a basis for further toxicity studies on M. rosenbergii and other potential tropical species.

by Fong Man Kim. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 127-137). / Abstracts in English and Chinese. / Thesis committee --- p.i / Acknowledgements --- p.ii / Abstract --- p.iii / List of figures --- p.viii / List of tables --- p.xi / Abbreviations --- p.xii / Table of contents --- p.xiv / Chapter CHAPTER 1 --- GENERAL INTRODUCTION --- p.1 / Chapter CHAPTER 2 --- LITERATURE REVIEW --- p.3 / Chapter 2.1 --- MIH from Penaeus vannamei --- p.3 / Chapter 2.1.1 --- General Introduction to P. vannamei --- p.3 / Chapter 2.1.1.1 --- Morphology --- p.3 / Chapter 2.1.1.2 --- Geographical distribution --- p.5 / Chapter 2.1.1.3 --- Economic value --- p.5 / Chapter 2.1.2 --- Physiology of Molting in Crustacean --- p.7 / Chapter 2.1.2.1 --- The molt cycle --- p.7 / Chapter 2.1.2.2 --- Physiological effects of ecdysone --- p.8 / Chapter 2.1.2.3 --- Regulation of the secretion of ecdysone --- p.9 / Chapter 2.1.2.4 --- Physiological effects of Molt-inhibiting hormone --- p.10 / Chapter 2.1.3 --- Cloning of MIH cDNA from P. vannamei --- p.14 / Chapter 2.1.3.1 --- Molecular identity of MIH --- p.14 / Chapter 2.1.3.2 --- Cloning of MIH cDNA --- p.15 / Chapter 2.1.3.3 --- Comparison of the cloned MIH-like cDNA with the CHH/MIH/VIH peptide family --- p.16 / Chapter 2.2 --- Plants as Bioreactors --- p.20 / Chapter 2.2.1 --- Principles & Techniques --- p.20 / Chapter 2.2.2 --- Advantages of plant bioreactors --- p.21 / Chapter 2.2.3 --- Tobacco expression system --- p.22 / Chapter 2.2.3.1 --- Tobacco as model plants --- p.22 / Chapter 2.2.3.2 --- Transformation methods --- p.23 / Chapter 2.2.4 --- Phaseolin --- p.26 / Chapter CHAPTER 3 --- EXPRESSION OF MIH IN TRANSGENIC TOBACCO --- p.28 / Chapter 3.1 --- Introduction --- p.28 / Chapter 3.2 --- Materials & Methods --- p.29 / Chapter 3.2.1 --- Chemicals --- p.29 / Chapter 3.2.2 --- Plant materials --- p.29 / Chapter 3.2.3 --- Bacterial strains and plasmid vectors --- p.30 / Chapter 3.2.4 --- Construction of chimeric genes - --- p.30 / Chapter 3.2.4.1 --- PCR amplification of MIH --- p.30 / Chapter 3.2.4.2 --- Cloning of PCR-amplified MIH into vector pET --- p.31 / Chapter 3.2.4.3 --- Cloning of MIH into vector pBK/Phas-sp and pTZ/Phas --- p.31 / Chapter 3.2.4.4 --- Cloning of MIH into binary vector pBI121 --- p.32 / Chapter 3.2.5 --- Transformation of Agrobacterium with pBI121/Phas-sp-MIH and pBI121 /Phas-MIH by electroporation --- p.39 / Chapter 3.2.6 --- Transformation of tobacco --- p.40 / Chapter 3.2.7 --- Selection of transgenic plants --- p.41 / Chapter 3.2.8 --- GUS assay --- p.42 / Chapter 3.2.9 --- Extraction of leaf genomic DNA --- p.43 / Chapter 3.2.10 --- Extraction of total RNA from developing seeds --- p.44 / Chapter 3.2.11 --- Synthesis of DIG-labeled DNA and RNA probes --- p.45 / Chapter 3.2.12 --- Southern blot analysis of genomic DNA --- p.47 / Chapter 3.2.13 --- Reverse transcriptase - polymerase chain reaction (RT-PCR) --- p.47 / Chapter 3.2.14 --- Northern blot analysis of total RNA --- p.48 / Chapter 3.2.15 --- Protein extraction and tricine-SDS-PAGE --- p.49 / Chapter 3.2.16 --- Purification of 6xHis-tag proteins --- p.50 / Chapter 3.2.17 --- Western blot analysis --- p.50 / Chapter 3.2.18 --- In vitro transcription & translation --- p.52 / Chapter 3.2.18.1 --- Construction of transcription vector containing the chimeric MIH gene --- p.52 / Chapter 3.2.18.2 --- In vitro transcription --- p.56 / Chapter 3.2.18.3 --- In vitro translation --- p.56 / Chapter 3.2.19 --- Particle bombardment --- p.57 / Chapter 3.2.19.1 --- Construction of MIH-GUSN fusion chimeric genes --- p.57 / Chapter 3.2.19.2 --- Conditions of particle bombardment --- p.63 / Chapter 3.2.20 --- Codon modification of MIH gene --- p.63 / Chapter 3.3 --- Results --- p.73 / Chapter 3.3.1 --- Construction of chimeric MIH genes --- p.73 / Chapter 3.3.2 --- "Tobacco transformation, selection and regeneration" --- p.73 / Chapter 3.3.3 --- Detection of GUS activity --- p.74 / Chapter 3.3.4 --- Southern blot analysis --- p.79 / Chapter 3.3.5 --- Detection of MIH transcript in transgenic tobacco --- p.83 / Chapter 3.3.5.1 --- RT-PCR --- p.83 / Chapter 3.3.5.2 --- Northern blot analysis --- p.86 / Chapter 3.3.6 --- Detection of MIH protein by Tricine-SDS-PAGE --- p.86 / Chapter 3.3.7 --- Detection of MIH protein by western blot analysis --- p.88 / Chapter 3.3.7.1 --- Western blot analysis using Anti-MIH antibody --- p.88 / Chapter 3.3.7.2 --- Western blot analysis using Anti-His antibody --- p.90 / Chapter 3.3.7.3 --- Western blot analysis using Anti-MIHA & Anti-MIHB antibodies --- p.90 / Chapter 3.3.8 --- Purification of 6xHis-tag proteins by Ni-NTA column --- p.94 / Chapter 3.3.8.1 --- Western blot analysis of proteins purified by Ni-NTA column --- p.97 / Chapter 3.3.9 --- In vitro transcription and translation --- p.100 / Chapter 3.3.9.1 --- In vitro transcription --- p.100 / Chapter 3.3.9.2 --- In vitro translation --- p.100 / Chapter 3.3.10 --- Particle bombardments --- p.103 / Chapter 3.3.10.1 --- Transient expression of MIH in soybean & tobacco leaves --- p.103 / Chapter CHAPTER 4 --- DISCUSSION --- p.107 / Chapter 4.1 --- Transient expression of MIH genes --- p.109 / Chapter 4.1.1 --- In vitro transcription and translation --- p.109 / Chapter 4.1.2 --- Particle bombardments --- p.220 / Chapter 4.2 --- Post-transcriptional gene silencing (PTGS) --- p.114 / Chapter 4.2.1 --- Post-transcriptional cis-inactivation --- p.114 / Chapter 4.2.2 --- Post-transcriptional trans-inactivation --- p.116 / Chapter 4.2.3 --- MIH gene and PTGS --- p.118 / Chapter 4.3 --- Codon usage --- p.119 / Chapter 4.3.1 --- Codon usage of MIH in plants --- p.120 / Chapter 4.3.2 --- Codon modification of MIH and further study on MIH expression in plants --- p.122 / Chapter 4.4 --- Post-translational protein degradation --- p.123 / Chapter 4.4.1 --- Construction of LRP-MIH fusion proteins --- p.123 / CONCLUSION --- p.125 / REFERENCES --- p.127

No description available.

Copy of author's previously published article inserted. / Bibliography: leaves 198-217. / 249 leaves : ill., maps ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / This thesis explores the impact of sustained disturbances on the ecology of algal dominated biofilms in the Lower River Murray, South Australia. It focuses on the physical effects of regulation through changes to the light environment and water level regime, and the local effects of grazing. The nutritional signficance of biofilms for the abundant populations of prawns and shrimps in the Lower Murray is also examined. / Thesis (Ph.D.)--University of Adelaide, Dept. of Zoology, 1997?

x, 53 leaves : ill., map ; 29 cm Notes Typescript Thesis (M.S.)--University of Oregon, 1986 Includes vita and abstract Bibliography: leaves 48-53 Another copy on microfilm is located in Archives

In this Master thesis the behaviour of anemone shrimp Ancylomenes longicarpus was studied in situ in the Red Sea. The diurnal activity, signalling behaviour and also cleaning behaviour was observed and analyzed.

The spatial and temporal patterns in the population demographs and feeding ecology of the caridean shrimp, Palaemon peringueyi, was investigated over a 12 month period in the permanently open Kariega Estuary situated on the south-east coastline of South Africa. Shrimps were sampled monthly from three stations corresponding to the lower, middle and upper reaches of the Kariega Estuary. The shrimps were collected at the different stations using a pull net. Temperature, salinity, dissolved oxygen and % cover of submerged vegetation were measured at each sampling station. Finally, the feeding ecology of the shrimp was investigated employing stable carbon (δ¹³C) and nitrogen (δ¹⁵N) isotopes analysis. Temperature, salinity, dissolved oxygen and vegetation cover showed distinct seasonal patterns. The shrimps attained total abundances and biomass of between 0 and 245.6 individuals per m² (ind.m⁻²) and between 0 and 13.6 grams wet weight per m² (g wwt.m⁻²), respectively. Mean abundance ranged between 2.3 (± 3.0) and 88 (± 27.3) ind.m⁻² in the lower reach, between 0.9 (± 0.4) to 21.5 (± 4.5) ind.m⁻² in the middle reach and 0 to 1.6 (± 0.5) ind.m⁻² in the upper reach. Mean biomass ranged between 0.4 (± 0.5) and 4.4 (± 1.4) g wwt.m⁻² in the lower reach, between 0.1 (± 0) to 4.3 (± 0.9) g wwt.m⁻² in the middle reach and between 0 to 1.6 (± 0.5) g wwt.m⁻² in the upper reach. Greatest abundances and biomasses were recorded in winter following heavy rainfall within the catchment area. Freshwater inflow which is due to rainfall is thought to provide a cue for the recruitment of marine breeding organisms. The spatial patterns in the distribution of the shrimps showed that juveniles occupied the lower and middle reaches and adults the entire length of the estuary. There were no significant relationships between the abundance and biomass of the shrimps and the selected physico-chemical variables (p > 0.05 in all cases). Results of the stable isotope study indicated that the mean δ¹³C of the shrimps in the lower, middle and upper reaches were -12.4 (± 1.3) ‰, -15.6 (± 0.5) ‰ and -17.5 (± 0.5) ‰, respectively. The δ¹⁵N isotopes of the shrimps in the lower reach was 11.6 (± 0.5) ‰ with middle and upper reaches having similar δ¹⁵N values, 13.5 (± 0.4) ‰ and 13.4 (± 0.4) ‰, respectively. There was no significant difference in the δ¹³C and δ¹⁵N isotope (p > 0.05) between the sexes of the shrimps. Two-way ANOVA indicated a significant difference in the δ¹³C isotope of the shrimps between the sites (p < 0.05). The difference in the δ¹³C and δ¹⁵N ratios of the shrimps suggests spatial variability in the diet of P. peringueyi within the Kariega Estuary. The diet of the shrimps in the lower reach is primarily composed of Nanozostera capensis epibionts. In the middle reach the diet of the shrimps is likely comprised of the detritus of benthic algae. In the upper reach of the estuary, P. peringueyi likely derives its carbon from a combination of epibionts, benthic algae and detritus derived from Codium.