Thesis (Ph.D.)--Ohio University, August, 2008. / Title from PDF t.p. Includes bibliographical references.

Thesis (Ph. D.)--New York University. / Bibliography: p. 227-229.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1932. / Typescript. With this is bound: The ovarian cycle in the viviparous teleost Xiphophorus helleri / Ralph J. Bailey. Reprinted from Biological bulletin, vol. LXIV, no. 2 (April 1933), p. 206-225. Includes bibliographical references (leaves 53-56).

Thesis (Ph. D.)--University of Chicago, 1922. / "Private edition, distributed by the University of Chicago libraries, Chicago, Illinois." "Reprinted from Biological bulletin, vol. XLV, no. I, July, 1923." Bibliography: p. 75-77. Also available on the Internet.

Das Melanom ist eine der aggressivsten Formen von malignen Tumoren beim Menschen. Bei Fischen der Gattung Xiphophorus kommt es zur spontanen Tumorformation, welche auch durch zwischenartliche Kreuzung herbeiführbar ist. Hybride mit angeborenem Melanom stellen ein nützliches Tiermodell zur Untersuchung der genetischen Grundlage der Tumorentwicklung dar. Ihre Tumorigenese hängt mit der pigmentzellspezifischen Überexpression der durch eine Mutation aktivierten Rezeptortyrosinkinase Xmrk zusammen. In reinrassigen Fischen wird die onkogene Funktion des xmrk durch den Genlocus R, welcher molekular noch nicht identifiziert wurde, unterdrückt. Zusammen mit der Überexpression von xmrk konnten mittels einer RNA-Seq Analyse weitere Gene gefunden werden, welche differenziell in den Proben von malignen und benignen Geweben des Xiphophorus exprimiert werden. Des Weiteren ist bekannt, dass die Methylierung des xmrk Promotors Einfluss auf die Expression des Genes hat. Um die Daten der durch RNA-Seq gefundenen Kandidatengene zu validieren, wurde deren Expression in malignen und benignen Geweben der Flossen und des Rumpfes mittels qPCR quantifiziert. Zusätzlich dazu wurde die Expression einiger humaner Orthologe dieser Gene in Proben aus humanen Melanomzelllinien gemessen. Mir war es möglich zu zeigen, dass mit Ausnahme von cdkn2ab, mitfb und xirp2b alle Kandidatengene signifikant unterschiedlich in mindestens einem Vergleich von benignem und malignem Gewebe exprimiert waren. Das mit xmrk verglichen gegensätzliche Expressionsmuster von pdcd4a macht es zu einem vielversprechenden Kandidaten als vom R-Locus codierten Tumorsuppressorgen. In den humanen Melanomzelllinien konnte ausschließlich von PDGFRB keine erhöhte Expression in irgendeiner Probe nachgewiesen werden. Während die Expression von PDCD4, C-MYC und MITF in mindestens drei der vier Zelllinien mittelstark erhöht war, ließ sich bei KIT eine enorm gesteigerte Überexpression in Zellen der Linie Hermes3a nachweisen. Da drei der fünf analysierten Gene und ihre Orthologen ähnliche Expressionsmuster in Proben des Xiphophorus und der humanen Melanomzelllinien zeigen, deuten diese Ergebnisse auf die Nützlichkeit des Tiermodells zur Identifizierung entscheidender Gene und Signalwege im malignen Melanom hin. Ein zweites Ziel der Arbeit war das Erlangen tieferer Einblicke in die Methylierung des Xiphophorus Melanoms auf einer globalen und promotor- spezifischen Ebene. Um die Hypothese einer Reduzierung der globalen Methylierung zu testen, führte ich eine kolorimetrische Quantifizierung der 5-mC DNA in Kontroll- und Tumorgeweben aus. Diese Vorgehensweise zeigte zum ersten Mal eine signifikante Verminderung der methylierten globalen DNA in den benignen Läsionen und malignen Melanomen der Flossen verglichen mit dem Kontrollgewebe. Um herauszufinden, on diese Demethylierung direkt mit der Überexpression des xmrk verbunden ist, analysierte ich als nächstes die Methylierung eines CpG Dinukleotids des xmrk Promotors mithilfe von methylierungssensitiven Restriktionsendonukleasen. Obwohl nur in den Proben des exophytischen Tumorwachstums als Krebsgewebe eine verringerte Methylierung des CpG Dinukleotids verglichen mit den Kontrollen nachgewiesen werden konnte, zeigte sich die Stelle in Zellen der Xiphophorus Melanomzelllinie PSM komplett unmethyliert. Diese Ergebnisse deuten stark daraufhin, dass eine differenzierte Methylierung das onkogene Potential dieser Zellen bewirkt. Um die Effekte veränderter globaler und promotor-spezifischer Methylierung auf die Tumorigenese besser zu verstehen, sind weitere Untersuchungen nötig. / Melanoma is among the most aggressive forms of malignant tumors in humans. In fish of the genus Xiphophorus melanoma tumor formation happens spontaneously in nature and can also be induced by interspecific crossing. Hybrid fish with hereditary melanoma are an established animal model for the study of the genetic origin of tumor development. Their tumorigenesis is linked to the overexpression of the mutationally activated receptor tyrosine kinase Xmrk in pigment cells. In purebred fish the molecularly still unrevealed locus R is suppressing the oncogenic function of xmrk. Along with the overexpression of xmrk a RNA-Seq analysis revealed even more differentially expressed genes in the tissues of malignant melanoma in Xiphophorus compared to benign tissues. Furthermore, there has already been gained evidence that the methylation status of the xmrk promotor has effects on its overexpression. To validate the RNA-Seq data of the candidate genes, gene expression in malignant and benign tissues of the fins and trunk was quantified using qPCR. Additionally, the expression of some human orthologues of these genes was also analyzed in samples of human melanoma cell lines. I was able to demonstrate that with the exception of cdkn2ab, mitfb and xirp2b all candidate genes are significantly differentially expressed in at least one set of tissues varying in dignity. The opposite expression pattern of pdcd4a compared to xmrk makes it a promising candidate as the at the R locus encoded tumor suppressor gene. In the human melanoma cell lines only the expression of PDGFRB wasn't increased in any of the samples. While the expression of PDCD4, C-MYC and MITF was moderately higher in at least three of the four cell lines, KIT was shown to be hugely overexpressed in Hermes3a. As three of the five analyzed genes and its orthologues show a similar expression pattern in samples of the Xiphophorus and the human melanoma cell lines, these findings point out the usefulness of the animal model to find new genes and pathways within the malignant melanoma. A second aim of the thesis was to gain a deeper insight into to methylation regulation in the Xiphophorus melanoma on a global and a promoter-specific level. To test the hypothesis that global methylation is reduced in the melanoma cells, I performed a colorimetric quantification of 5-mC DNA in control and tumor tissues. This approach showed for the first time a significantly decreased amount of global DNA in the benign and malignant samples deriving from the fins compared to control tissues. To find out if this demethylation is directly linked to the overexpression of xmrk, I analyzed the methylation of CpG site in the xmrk promotor using methylation sensitive restriciton endonucleases. Interestingly in the samples of the Xiphophorus melanoma cell line PSM the CpG site wasn't methylated at all. Although only the samples of the exophytic tumor growth as a tumoric tissue were less methylated than the control, the cells of the Xiphophorus melanoma cell line PSM were completely unmethylated. These results imply that differential methylation triggers the oncogenic potential of these cells. To improve the understanding of the effects global and promoter-specific methylation has on tumorigenesis, further studies are necessary.

Swordtail fishes of the genus Xiphophorus have been studied as a model of sexual selection for many years. Many single-trait manipulation studies have been performed, determining female preferences for individual male traits. I characterized how five traits (standard body length, body depth, dorsal fin width, sword length and vertical bar number) correlate to one another within natural variation of populations of X. birchmanni, X. malinche and three hybrid populations and created synthetic 3- dimensional animations exhibiting these traits within ranges of natural variation. I then performed choice tests on females of the above populations using a computer system that automatically played these stimulus videos and simultaneously tracked a female’s position within a test tank to determine female preference for different male phenotypes. Only X. birchmanni females showed significant preferences. Their preferences were in line with past research of univariate trait manipulation experiments. They showed significant preference for larger bodies and dorsal fins and smaller or no swords. They also showed a non-significant preference for vertical bar numbers. My results also confirmed univariate studies in which X. malinche females showed reduced preference for conspecific males and being rather indifferent to the presence of swords. Hybrid females were also shown to have reduced preferences for any specific trait, suggesting that they express recombinant preferences, which can also be explained by reduced color vision at low levels of light.

Thesis (M.S.)--Ohio University, November, 2009. / Release of full electronic text on OhioLINK has been delayed until September 1, 2010. Title from PDF t.p. Includes bibliographical references.

The aim of this work was to investigate the nutrition of a popular ornamental fish, the swordtail (Xiphophorus helleri), under intensive aquaculture conditions. The study focused on the formulation and manufacture of artificial dry diets, as well as the influence of livefeed supplementation on growth and nutritional indices of both juvenile and broodstock fish. A combination of 65% fishmeal and 35% soya oil cake meal was found to closely approximate the essential amino acid profile of X helleri (r² = 94,3%) and lysine was identified as the first limiting amino acid. The influence of dietary protein and energy on the growth of X helleri was investigated by means of a growth trial comparing a range of nine test diets combining 3 crude protein (45,38 and 30%) and 3 digestible energy (16,5, 14,5 and 12,5 kJ/gram) levels. A high crude protein content (45% dry matter) and a protein to energy ratio of ≥ 27 mg protein per kJ digestible energy was shown to promote optimal growth rate and feed conversion ratio in juvenile X helleri. The final body protein and lipid content of the fish were significantly correlated with the dietary protein to energy ratio. The nutritional value of identical dietary formulations prepared by drum-drying or cold extrusion techniques and presented either as flake or crumble particles was evaluated in a growth trial and through analysis of vitamin C levels in the diets. Up to 27% of vitamin C activity was destroyed during the drum-drying of flake diets, while 80% of the remaining vitamin C was lost through leaching within 10 minutes of immersion in water. A crumble-type diet was shown to be acceptable to X helleri, and yielded significantly better growth rate and feed conversion than the equivalent flake feed. Almost 19% of fish fed the flake diets developed vitamin C deficiency symptoms including scoliosis and lordosis, while this was completely prevented by feeding crumbles, indicating a significant restriction of leaching losses from this type of particle. The use of livefeed (Daphnia spp.) as a daily supplement to dry feeds was shown to result in a significant improvement in growth rate and survival of juvenile swordtail fish. Furthermore, the synergistic action of nutrients contained in the livefeed resulted in an improvement in the feed conversion efficiency of the artificial formulation. While mortalities of 13,4% and 15% were recorded in fish fed exclusively on flake feed or Daphnia, no mortality occurred in the group fed the supplemented diet. Furthermore, the incidence of vitamin C deficiency symptoms was prevented by daily supplementation with Daphnia. Fish fed exclusively on Daphnia exhibited significantly more intense pigmentation due to carotenoids contained in the livefeed. In broodstock fish maintained primarily on flake diets, growth rate, feed conversion ratio as well as reproductive performance, as measured by fecundity, were significantly improved by daily livefeed supplementation. Weekly supplementation showed no measurable advantage over flake feed only. A direct correlation was demonstrated between fecundity and size of female fish with a mean fecundity of 190 embryos per female in those fed a daily Daphnia supplement. This was significantly higher than the yield from the non-supplemented or weekly supplemented treatments (133 and 140 embryos per female respectively). Despite these differences, the number of young harvested during the experiment did not differ between treatments and this discrepancy was ascribed to parental cannibalism of newborn juveniles. This phenomenon constitutes a major potential bottleneck in production with estimates of up to seventy percent of young lost in some exp experimental tanks. In conclusion, the present study demonstrated that established aquaculture nutrition principles and techniques are applicable to the tropical ornamental fish species, X helleri. It was demonstrated that by using balanced crumble feed formulations, and regular livefeed supplementation, production yields under intensive conditions may be significantly improved. Suggestions for future work on poeciliid production in South Africa include investigation of the economic implications of the recommendations stemming from this project, and further research into effective restriction of parental cannibalism of newborn fish.

Thesis (M.S.)--Ohio University, August, 2007. / Title from PDF t.p. Includes bibliographical references.

Adult fish that belong to the family Poeciliidae cannibalise juveniles, both in the wild and under captive conditions, but this behaviour has only been partly investigated in the Poeciliidae in some of the commercially valuable species. The objective of the research is to develop an understanding of intercohort cannibalism and parturition-associated behaviour in captive-bred swordtail (Xiphophorus helleri), with applications to industry and future research of other poeciliids. Experiments investigating the effect of adult stocking density and sex ratio on the production of juveniles were used to determine if cannibalism occurs under culture conditions. The average rate of intercohort cannibalism ranged from (5.5 to 53.9%), and was positively density dependent and independent of sex ratio, indicating that males and females were probably equally cannibalistic. The highest number of juveniles (1725.7±141.4) produced per tank over 70 days was obtained from two males and eight females. To develop a better understanding of adult and juvenile behaviour during parturition, fish were observed under controlled laboratory conditions using video and behaviours such as attack (burst of speed by an adult in the direction of a juvenile), escape (avoidance of cannibalism after attack) and cannibalism (predation of a live juvenile by an adult), for example, were identified. Under laboratory conditions most young escaped in downward direction after attack (49%) and most utilised the refuge made that was made available. Furthermore, most attacks (62-65%) and cannibalism (57-84%) occurred at the bottom. Since the presence of refuge significantly increased the rate of juvenile survival under culture conditions, it was hypothesised that the rate of cannibalism could be reduced under farming conditions if juveniles were protected when they escaped downwards. This hypothesis was accepted as it was found that refuge at the bottom of the water column or the inclusion of a false-bottom reduced the rate of cannibalism by 49% and 72%, respectively. Similarly, the hypothesise that the rate of cannibalism could be reduced if juveniles where given protection when escaping sideways (32% of juveniles escaped sideways in the laboratory) was also accepted when tested under farm-scale conditions because a false-side reduced the rate of cannibalism by an average of 45%. Since males and females were equally responsible for cannibalising juveniles in the laboratory, it was hypothesised that the rate of cannibalism would decrease proportionately with the removal of males (Le. 20% of the cannibals) from the population; the removal of males under farming conditions resulted in a 19.5% reduction in the rate of cannibalism. Since older juveniles were better able to escape cannibalism than neonates and since adults habituate to stimuli that previously resulted in attack behaviour, it was hypothesised that the rate of cannibalism would remain unaffected by the length of time that juveniles were exposed to adults in the breeding tanks. This hypothesis was also accepted when tested under farm conditions. However, some hypotheses based on laboratory observations were not accepted. For example, a constant low light intensity did not appear to decrease the rate of cannibalism under farm conditions; also, the occurrence of dead and deformed juveniles went unnoticed in the laboratory, and under farm conditions, where adults did not have access to the bottom of the tank, 10% of the harvest consisted of dead and deformed juveniles. It is concluded that technologies, such as bottom-refuge or a false-side, that increase the size of the liveharvest and allow for the removal of potentially less viable offspring are recommended for the commercial production of poeciliids. The overall similarity of X. helleri behaviour between the laboratory experiments and the farm-scale trials suggests that the post-partum behaviour of X. helleri remains consistent under these different conditions; thus, behaviour under one set of conditions may be used to predict behaviour under other conditions. The application and significance of extrapolations to industry and future research of X. helleri and possibly other poeciliids were discussed and the most applicable laboratory observations with the highest extrapolation capacity were proposed. Furthermore, techniques were developed to aid industry and future researchers in making predictions relating to behaviour of X. helleri under different conditions based on laboratory observations. The results were used to develop a model indicating that selection pressures against cannibalism are not likely to exist at the rate of cannibalism observed here since the potential genetic gain through kin survival and inclusive fitness was shown to be greater than any potential genetiC loss experienced by a victim of cannibalism. The model was successfully tested under a range of social conditions. Other possible explanations for cannibalism in poeciliids, such as parental manipulation, nutritional advantages, opportunistic predation and the recovery of energy are discussed. It is suggested that the most likely proximate cause of cannibalism under captive conditions is opportunistic predation. The theory that cannibalism ensures that only viable genes of the victim are expressed, through inclusive fitness, is a possible ultimate cause of cannibalism, which may have been inherited from feral ancestors of captive-bred X. helleri.