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search.filters.applied.f.advisor: Snell, Terry × End date: 2019 × Start date: 2016 × search.filters.applied.f.isOrgUnitOfPublicationTitle: College of Sciences × search.filters.applied.f.isSeriesOfPublicationTitle: Undergraduate Research Option Theses ×

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Rotifer Growth Under Astaxanthin Enrichment

2017-12 , Siegfried, Emma

Rotifers and astaxanthin both play an important part in the aquaculture industry. Rotifers are used as a substitute for copepods, the main source of food for larval fish in natural systems, due to the ease with which they can be cultured. Astaxanthin is a carotenoid and antioxidant which brightens the coloring of fish and improves fish health. Rotifers are believed to be a method through which astaxanthin can be bioencapsulated and vectored to larval fish. As a result, it is important to understand the effect of astaxanthin on rotifers themselves. This experiment uses a multitude of different protocols to determine how different astaxanthin compounds effects rotifers on both the individual and population levels. Reproductive tables and fluorescent imaging were used to assess the health of individual rotifers; population density measurements in mass cultures were used to assess rotifer population health. The reproductive ability of rotifers was significantly different from control under multiple astaxanthin treatments. Astaxanthin enrichment also created a higher stable population density in the mass cultures. The fluorescent imaging showed that the rotifers reached peak astaxanthin concentration within the rotifer gut after 3 hours, and but concentration returned to control levels within 24 hours of removal from astaxanthin. These results all point to the fact that astaxanthin helps to increase rotifer health and fitness, and that these rotifers could be used as a vector for astaxanthin to larval fish.

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Establishing a Working Protocol for Plasmid Cloning and shRNA Design in Endogenous Brachionus manjavacas Gene TRP7

2016-07-18 , Krishnappan, Sharadha

Current transfection protocol in rotifers only allows for temporary transfection within rotifers and does not allow for the continuous knockdown of endogenous genes, thereby inhibiting the possibility of observing long-term biological effects in response to specific perpetual gene knockdowns. This study aims to address this particular issue by establishing a working protocol for plasmid cloning and shRNA design within an endogenous gene of B. manjavacas with known biological effects, allowing for the exploration into the optimization of a transfection protocol and demonstration of RNAi knockdown of the known gene within the rotifers as subsequent studies. Manipulation of gene expression in rotifers could occur through plasmid vector insertions, which induce silencing of a gene’s expression with short hairpin RNA (shRNA), via RNAi. This would effectively stimulate gene knockdown, allowing for the observation of biological effects such as changes in fecundity and lifespan. With the establishment of a working protocol for plasmid cloning and shRNA design, as a result of this study, the optimization of a transfection protocol for rotifers is explored. With increased efficiency in the transfection of rotifers, populations of rotifers expressing the plasmid can be amassed, allowing for experimental design that examine the varying aging mechanisms and effects that are stimulated due to permanent changes in target gene expression through RNAi. This, in turn, could give rise to the identification of evolutionarily conserved genes that regulate organismal aging, which could lead to further implications in the field of pharmacological intervention in mammalian aging as well as in the field of biogerontology overall.

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