Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O gênero Synbranchus, principal grupo representante da família Synbranchidae no Brasil é composto por três espécies: Synbranchus marmoratus Bloch, 1795, com distribuição do Norte da Argentina até o México; Synbranchus madeirae Rosen e Rumney, 1972, encontrado no Rio Madeira, Bacia Amazônica; e Synbranchus lampreia Favorito et al., 2005, encontrada no Rio Goiapi, Ilha de Marajó. Após a descrição de S. marmoratus (primeira espécie descrita para o gênero) outras doze espécies foram descritas para o grupo até o início do século XX. Em função da imprecisão das descrições e/ou ausência de holótipos conhecidos, Günther (1870) e Ringuelet (1967) colocaram-as em sinonímia com S. marmoratus. Incrédulos da distribuição geográfica tão ampla da espécie Norte da Argentina até o México e fundamentados na imprecisão das descrições, trabalhos posteriores verificaram a presença de outras espécies sobrepondo certos sítios de distribuição da espécie Synbranchus marmoratus. Como resultado, foram descritas as espécies Synbranchus madeirae e Synbranchus lampreia, além de uma nova espécie para o gênero Ophisternon, O. aenigmaticum Rosen e Greenwood, 1976. Ainda hoje, acredita-se que a espécie Synbranchus marmoratus seja, na realidade, formada por um complexo de espécies (ROSEN e GREENWOOD, 1976). Em função da escassez de estudos osteológicos completos e de caracteres diagnósticos bem definidos, as espécies do gênero Synbranchus são difíceis de serem distinguidas e a espécie S. marmoratus é frequentemente confundida com a espécie O. aenigmaticum. Além disso, trabalhos anteriores (FAVORITO-AMORIM, 1992, 1998) revelaram a existência de espécimes do gênero Synbranchus que não possuem algumas características diagnósticas para esse gênero. O presente estudo apresentou, pela primeira vez, a osteologia completa da espécie Synbranchus marmoratus, comparou exemplares dessa espécie oriundos de diferentes localidades e examinou alguns dos caracteres diagnósticos definidos para o gênero Synbranchus e suas espécies. Não foi possível encontrar diferenças significativas entre os espécimes provenientes de diferentes localidades, que justificasse a separação destes em espécies distintas. O resultado mostrou que alguns dos caracteres diagnósticos para o gênero Synbranchus não são exclusivos para esse grupo ou são extremamente variáveis entre os espécimes, reiterando a necessidade de uma rediagnose para esse gênero, anteriormente proposta por Favorito-Amorim (1992,1998) e Favorito et al. (2005). O resultado do exame dos caracteres diagnósticos das espécies do gênero Synbranchus apresentou um resultado semelhante, revelando que diversas características utilizadas para a distinção das espécies são, na realidade compartilhadas ou sobrepõem-se. Os resultados sugerem que a descrição completa da osteologia se S. marmoratus revelou-se uma importante ferramenta taxonômica e mostrou que faz-se necessária a realização de estudos osteológicos envolvendo as demais espécies do gênero Synbranchus.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O gênero Synbranchus, principal grupo representante da família Synbranchidae no Brasil é composto por três espécies: Synbranchus marmoratus Bloch, 1795, com distribuição do Norte da Argentina até o México; Synbranchus madeirae Rosen e Rumney, 1972, encontrado no Rio Madeira, Bacia Amazônica; e Synbranchus lampreia Favorito et al., 2005, encontrada no Rio Goiapi, Ilha de Marajó. Após a descrição de S. marmoratus (primeira espécie descrita para o gênero) outras doze espécies foram descritas para o grupo até o início do século XX. Em função da imprecisão das descrições e/ou ausência de holótipos conhecidos, Günther (1870) e Ringuelet (1967) colocaram-as em sinonímia com S. marmoratus. Incrédulos da distribuição geográfica tão ampla da espécie Norte da Argentina até o México e fundamentados na imprecisão das descrições, trabalhos posteriores verificaram a presença de outras espécies sobrepondo certos sítios de distribuição da espécie Synbranchus marmoratus. Como resultado, foram descritas as espécies Synbranchus madeirae e Synbranchus lampreia, além de uma nova espécie para o gênero Ophisternon, O. aenigmaticum Rosen e Greenwood, 1976. Ainda hoje, acredita-se que a espécie Synbranchus marmoratus seja, na realidade, formada por um complexo de espécies (ROSEN e GREENWOOD, 1976). Em função da escassez de estudos osteológicos completos e de caracteres diagnósticos bem definidos, as espécies do gênero Synbranchus são difíceis de serem distinguidas e a espécie S. marmoratus é frequentemente confundida com a espécie O. aenigmaticum. Além disso, trabalhos anteriores (FAVORITO-AMORIM, 1992, 1998) revelaram a existência de espécimes do gênero Synbranchus que não possuem algumas características diagnósticas para esse gênero. O presente estudo apresentou, pela primeira vez, a osteologia completa da espécie Synbranchus marmoratus, comparou exemplares dessa espécie oriundos de diferentes localidades e examinou alguns dos caracteres diagnósticos definidos para o gênero Synbranchus e suas espécies. Não foi possível encontrar diferenças significativas entre os espécimes provenientes de diferentes localidades, que justificasse a separação destes em espécies distintas. O resultado mostrou que alguns dos caracteres diagnósticos para o gênero Synbranchus não são exclusivos para esse grupo ou são extremamente variáveis entre os espécimes, reiterando a necessidade de uma rediagnose para esse gênero, anteriormente proposta por Favorito-Amorim (1992,1998) e Favorito et al. (2005). O resultado do exame dos caracteres diagnósticos das espécies do gênero Synbranchus apresentou um resultado semelhante, revelando que diversas características utilizadas para a distinção das espécies são, na realidade compartilhadas ou sobrepõem-se. Os resultados sugerem que a descrição completa da osteologia se S. marmoratus revelou-se uma importante ferramenta taxonômica e mostrou que faz-se necessária a realização de estudos osteológicos envolvendo as demais espécies do gênero Synbranchus.

Rivulus marmoratus (Cyprinodont; Aplochelidae) is the only vertebrate known to reproduce by obligate internal self-fertilization. Selfing is the ultimate form of inbreeding and leads to virtual homozygosity in approximately ten generations, so that this mode of reproduction is effectively clonal. Populations of this species consist of arrays of homozygous clones, with high clonal diversity and low representation of each clone. Males occur at low frequency (<1%) in most populations, with a few notable exceptions. Females have never been recorded.High frequencies of males have been reported on two occasions; high numbers of males were reported on the island of Curacao in the 1970's, and males comprised up to 24% of the populations sampled on several Belize Cays during 1990-91. Hermaphrodites collected from one island, Twin Cays, proved to be heterozygous at mini-satellite loci when progeny tested and DNA fingerprinted. This was evidence that recent outcrossing had occurred.Earlier experiments with Floridian clonal lines had suggested that sex in this species was thermolabile; incubation of embryos at low temperatures (19° C)) produced up to 100% males. It was suggested that this might be part of an environmental sex determination (ESD) system in this species.If the induction of males is indeed part of and ESD in this species, then it may be that the temperature at which males are induced in a particular clone will be related to the temperature regime that clone encounters in nature. Therefore, it was hypothesized that the temperature at which males are induced would be related to the geographic origin of each clone, so that clones from the cooler extremes of the range would produce males at a lower threshold temperature than clones from the more equatorial center of the range. This study used laboratory reared descendants of clones from Vero Beach, Florida, Rio de Janiero, Brazil (the extreme north and south of the range), and from several Belize barrier islands and the Belize mainland (near the center of the range) in an effort to detect differences in the production of males between clonal lines based on their geographic origin. Eggs were collected from these hermaphrodites and incubated at 26° C)C, 22.5° C), or 19 ° C), representing normal, and below average temperatures for this species. These embryos were hatched and reared to sexual maturity at which time their sex was evaluated.Incubation at 26° C) resulted in 2-17% males, incubation at 22.5° C) resulted in 6-53% males, and incubation at 19° C) resulted in 47-74% males. However, the variation in male induction between clonal lines did not correspond to a geographic hypothesis.Also, significant numbers of deformed individuals were noted, particularly among individuals incubated at the lower two temperatures. Incubation at 26° C) resulted in 0-24% deformed, incubation at 22.5° C) resulted in 6-66% deformed, and incubation at 19° C) resulted in 36-87% deformed. It is clear that development at these lower temperatures is difficult for this species, and it may be that low temperature developmental stress is not a general challenge to this species. / Master of Science

The mangrove killifish, Kryptolebias marmoratus, has the potential of becoming a strong model organism for a range of biological disciplines thanks to its ability to self-fertilise, a process only known to occur in invertebrate animals until its discovery. Selfing, a natural occurrence in this species, has lead to the formation of clonal lineages composed of highly homozygous individuals. The aim of this thesis was to further establish K. marmoratus in the field of developmental biology by providing an information infrastructure to help advance research on this peculiar animal and further promote its place in the pantheon of model organisms. To do so, I first set out to standardise K. marmoratus embryology by providing defined developmental stages with clear visual representations of key embryonic structures. This staging series is an essential tool that will ensure repeatability and consistency within and across different laboratories. Secondly, I examined several techniques for embryonic manipulation and for imaging that can be used in an array of experimental designs. Using these techniques I demonstrated microinjection of embryos by monitoring the yolk syncytial layer and its nuclei, and time-lapse analyses of the yolk surface during embryonic development. Finally, I applied the knowledge gained from my first two studies and examined Bmp signalling in K. marmoratus embryos and its influence on body patterning. By inhibiting this pathway, I found a new phenotype characterised by an extremely short and split body axis. These data highlighted the importance of studying known signalling pathways in unknown organisms as species-specific differences may improve our understanding of fundamental developmental processes. This thesis demonstrates that with its easily obtainable and manipulated embryos, K. marmoratus can be used for embryological research in the same light as other model organisms such as zebrafish or medaka. The rising amount of information on mangrove killifish will help further take advantage of this unique and intriguing species, and supports the use of this hermaphroditic vertebrate as a strong comparative model in developmental biology.

Thesis (Ph. D.)--Virginia Polytechnic Institute and State University, 1993. / Vita. Abstract. Includes bibliographical references (leaves 109-117). Also available via the Internet.

The ability of natural selection to promote and preserve genetic variation at Major Histocompatibility Complex (MHC) class I loci was examined in two fish species known to have low genetic variation either as a consequence of their breeding system or population structure. The tempo and modes of molecular evolution acting on these loci has also been discussed. The marine killifish Rivulus marmoratus is the only vertebrate known to exist in nature in homozygous form. The findings of this study suggest that MHC class I genes at one locus in R. marmoratus have rapidly accumulated variation, particularly in region of the gene encoding functional important domains. This rapid accumulation of variation is likely due to the increased intensity of natural selection acting on these genes resulting from the homozygosity of the species. The contention that the variation characterized at one MHC locus is of recent origin is further supported by an analysis of the mitochondrial control region if R. marmoratus, which suggests that the individuals included in this study are recently diverged. The variation characterized here is the first evidence of genetic variation at coding loci in this species, and may be evidence of the cost of homozygosity for R. marmoratus. Variation at a single MHC class I locus was also characterized in several members of the Death Valley pupfish species complex. Two of these species, Cyprinodon diabolis and Cyprinodon salinus have undergone intense historical or ongoing population bottlenecks. Despite the action of random genetic drift acting on these species, both maintain multiple MHC class I alleles at one locus. The alleles characterized in C. diabolis and C. salinus were also recovered from other members of the Death Valley complex. This suggests that overdominant natural selection has acted to maintain ancestral genetic variation at this locus in both species. This result has important implications for the design of conservation programs for these endangered species, and may have broader implications for the design of captive breeding programs of species with highly reduced population sizes in general. / Ph. D.

No description available.

Made available in DSpace on 2014-12-17T14:10:29Z (GMT). No. of bitstreams: 1 NirleiHCB_DISSERT.pdf: 6542994 bytes, checksum: b970fd5055cebafb782b426aa47838fa (MD5) Previous issue date: 2012-03-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Reproductive strategy of the hermaphrodite fish the marbled swamp eel, Synbranchus marmoratus (Bloch, 1795) (Osteichthyes: Synbranchidae) was investigated in the Marechal Dutra reservoir, Acari, Rio Grande do Norte, using fish samples captured monthly during the period of July 2010 to July 2011. The limnological parameters and rainfall were verified. The captured fish specimens were numbered, weighed, measured, dissected and their gonads and liver were removed, weighed and identified as to sex and developmental stages of gonads. The length-weight relationship was determined for grouped sex. The size of first gonadal maturation, gonadosomatic index (GSI), condition factor (K) and hepatosomatic index (HSI) were calculated. The development stages of the gonads were investigated using macroscopic and histological techniques, besides the fecundity and reproductive period. The macroscopic characteristics and the histological development of gonads confirmed the four different sexual types in the sampled population: primary males, females, intersex and secondary males. The secondary males were bigger and heavier than all the other types. This species showed a negative allometric growth with a higher increase in length than weight. The size at which 50% of the females began the process of gonadal maturation was 43.5 cm of total length, and for secondary males it was 58.5 cm. Condition factor showed two peaks in February and in April 2011. IGS indicated a reproductive period from July to September 2010 and was negatively correlated with the condition factor and rainfall. IGS showed a negative correlation when compared with the condition factor and the rainy season. HSI was highest in February ad lowest in December. The reproductive period of this species occurs between the end of the rainy season and the beginning of the dry season. S. marmoratus presents a seasonal reproductive strategy which optimizes its reproductive success under the semiarid climatic conditions / A estrat?gia reprodutiva do peixe hermafrodita mussum, Synbranchus marmoratus (Bloch, 1795) (Osteichthyes: Synbranchidae) foi investigada no a?ude Marechal Dutra, Acari, Rio Grande do Norte, utilizando os exemplares capturados mensalmente no per?odo de julho de 2010 a julho de 2011. Foram verificados os par?metros limnol?gicos e a pluviosidade. Os exemplares capturados foram numerados, pesados, medidos, dissecados e o f?gado e as g?nadas foram retiradas, pesadas e identificadas quanto ao sexo e o est?dio de desenvolvimento gonadal. A rela??o peso-comprimento foi determinada para sexos agrupados. O tamanho da primeira matura??o, o ?ndice gonadossom?tico (IGS), o fator de condi??o (K) e o ?ndice hepatossom?tico (IHS) foram determinados. As fases do desenvolvimento das g?nadas foram investigadas usando t?cnicas macrosc?picas e histol?gicas, al?m de determinar a fecundidade e o per?odo reprodutivo da esp?cie. As caracter?sticas macrosc?picas e histol?gicas de desenvolvimento gonadal confirmaram os quatro tipos sexuais diferentes na popula??o dos peixes capturados, sendo: machos prim?rios, f?meas, intersexos e machos secund?rios. Os machos secund?rios apresentaram os maiores valores de comprimento total e peso total. A esp?cie mostrou um crescimento do tipo alom?trico negativo com maior incremento em comprimento do que em peso. O tamanho em que 50% da popula??o das f?meas iniciaram o processo de matura??o gonadal foi de 43,5 cm de comprimento total e para os machos secund?rios foi de 58,5 cm. O fator de condi??o apresentou dois picos, um no m?s de fevereiro e outro m?s de abril de 2011. IGS indicou um per?odo de reprodu??o durante julho a setembro de 2010, mostrando correla??o negativa ao fator de condi??o. IHS mostrou um pico no m?s de fevereiro com o menor valor no m?s de dezembro coincidindo com o per?odo de seca da regi?o. A esp?cie apresenta um per?odo reprodutivo entre o final das chuvas e o in?cio da seca. S. marmoratus mostra uma estrat?gia reprodutiva sazonal que aperfei?oa seu sucesso reprodutivo dentro das condi??es clim?ticas do semi?rido. Palavras chave: mussum, Synbranchus marmoratus, estrat?gia reprodutiva, hermafroditismo, a?ude do semi?rido

It is widely documented that non-human organisms express individual differences in behavioural patterns. For example individuals can be categorised as bold or shy and when these individual behavioural differences are consistent through time, they are termed behavioural types (BTs). In recent years research has identified that BTs often correlate across contexts/situations and these correlations are referred to as behavioural syndromes. Behavioural types and syndromes (i.e. personality) have also been implicated as major factors shaping population dynamics and the ability to buffer environmental disturbance. Recent theoretical predictions have proposed that BT variation may be underpinned by life-history strategies; however, these predictions have been little studied to date. Moreover, little research has focused upon environmental influences and the ontogeny of personality. In this thesis I use the Mangrove killifish (Kryptolebias marmoratus), a naturally occurring clonal vertebrate, as a model organism. This species presents a powerful tool providing the ability to replicate within and between isogenic genotypes in a controlled manner. Moreover the natural clonality expressed by this species permits environmental effects upon BT plasticity and BT-life-history interactions to be investigated within a developmental framework. In chapter 2, I present microsatellite genotyping results which show that the founding individuals used to propagate a laboratory population at The University of Exeter represent 20 genetically distinct homozygous genotypes. I additionally address five research questions exploring genotypic, environmental, and developmental effects upon three commonly studied BTs (exploration, boldness and aggression): Firstly; I ask do adult hermaphrodite and secondary males exhibit personality i.e. repeatable BT expression? In chapter 3, I present results showing that both of the sexes express short term personality. Moreover, I show that that genotype is an important factor influencing BTs expressed, regardless of sex, indicating underlying genetic control. Secondly I ask; does genotype level life-history variation underpin personality trait variation during ontogeny? In chapter 4, I show considerable developmental plasticity in behavioural expression between genotypes but not life-history and I find limited behaviour-life-history relationships during development. Thirdly I ask; does the rearing environment influence life-history and behavioural plasticity? In chapter 5, I show that in comparison to a control treatment, the presence of conspecifics during ontogeny results in an average reduction in behavioural scores; however, life-history was unaffected. In addition, I show that development in a low food environment lowered average exploration and growth rate but had no effect on boldness or aggression. Furthermore, fish exposed to a predation risk simulation during ontogeny exhibited similar behavioural scores as the control, yet this treatment generated BTs i.e. personality. My fourth question asks; does the parental rearing environment (utilised in chapter 5) influence behavioural expression in the next generation? In chapter 6, I show that transgeneratonal effects of each parental rearing environment influenced life-history but had a minimal effect upon behaviour in the next generation. Finally I ask; does kin or familiarity influence plasticity in associations and aggression? In chapter 7, I show that genotypes have the ability to discriminate kin and familiars and modulate aggression and association accordingly. These results support the concept that developmental and environmental induced plasticity may be more important than life-history in shaping behaviour. Furthermore, although adults exhibit personality and genotypic effects appear important, genotype interacts with environmental/experiential influences to differentially shape behavioural plasticity during ontogeny. I suggest that theoretical predictions regarding life-history may be insufficient to explain the complexity of animal personality in this species. I discuss these results within developmental and epigenetic frameworks with reference to the ecological significance of these patterns within this species and the animal kingdom as a whole.

The self-fertilizing mangrove rivulus, Kryptolebias marmoratus, is an amphibious fish capable of reversible gill remodelling when moving between aquatic and terrestrial environments. In this thesis I determined how plastic morphological and physiological respiratory traits were integrated during transitions between environments. In two isogenic lineages, I found that behaviour (increased emersion) of individual fish caused gill morphological changes (enlargement of the interlamellar cell mass (ILCM)) that reduced gill surface area. I also found that large ILCMs that formed after 7 d of air exposure increased both gill ventilation and critical oxygen tension (Pcrit) when fish returned to water. These results indicate that large ILCMs reduce aquatic respiratory function, and increased gill ventilation was unable to maintain oxygen uptake at extreme levels of hypoxia. Ultimately, this study highlights the trade-offs in gill structure and function during the transition between air and water, and demonstrates that differences in behaviour can generate morphological variation.