Since their recent introductions into Florida waters, three nonnative species [Perna viridis Linnaeus, 1758 (Asian green mussel), Mytella charruana d'Orbigny, 1846 (charru mussel) and Megabalanus coccopoma Darwin, 1854 (pink titan acorn barnacle)] have expanded both north and south along the Atlantic coast. Very little research has been done to understand how these nonnative species interact with the native eastern oyster (Crassostrea virginica Gmelin, 1791), which is a keystone species that provides important ecological services and economic benefits. To test the potential effects of P. viridis, M. charruana and M. coccopoma on C. virginica, I addressed the following questions: 1a) Does the presence of nonnative species decrease oyster larval settlement? 1b) Do oyster larvae avoid settling on oyster shells to which nonnative species are attached? 2a) Do nonnative species decrease survival of juvenile oysters (spat)? and 2b) Do nonnative species hinder spat growth? My manipulative experiments showed that the tested nonnative species influenced settlement, growth and survival of C. virginica in unique ways. Megabalanus coccopoma decreased the total number of settled oyster larvae, but did not influence larval preference or survival and growth of spat. Perna viridis negatively influenced larval settlement and oyster larvae avoided settling on shells of P. viridis. Mytella charruana had no influence on the total number of settled larvae but oyster larvae avoided settling on oyster shell with M. charruana or on the mussel shells themselves. Furthermore, both nonnative mussels negatively affected the survival of juvenile oysters, but only M. charruana reduced spat growth. These three nonnative species should be classified as invasive species because all had negative effects on the native oyster C. virginica.

There have long been concerns about the negative impacts of recreational boating activity in the Indian River Lagoon system (IRL), especially in Mosquito Lagoon (ML), the northernmost part of the IRL. My research is focused on the impacts of boat wakes on intertidal reefs formed by the eastern oyster, Crassostrea virginica. There has been a 24% loss of oyster habitat in ML since 1943, where natural oyster reefs have been replaced by dead oyster reefs which do not serve the same ecological function. While there is anecdotal and correlative evidence that this loss is a result of boat wakes, no studies to date have confirmed dead reefs can be a direct result of boat wakes. Therefore, I addressed the following questions: (1) What wake heights are generated by a range of boat types, and (2) What amount of oyster movement and erosion occurs as a result of these boat wakes? A series of boat pass experiments addressed the first question; these results were utilized in experiments at Florida Institute of Technology's wave tank to observe sediment erosion and oyster movement as a result of specific wake heights. Model selection was used for both the field and wave tank experiments to determine which variables contributed most to explaining the wake heights, erosion, and oyster movement that occurred. Wake heights ranging from 0.05 cm to 20.80 cm were documented contacting the oyster reefs from the boat passes, with a mean of 2.95 cm. Boat type was less important than speed or distance when determining wake height. My wave tank results document that wake heights as small as 2 cm contacting oysters are capable of moving individual and clusters of oysters. Minimum distances for boats to travel in order to maintain wakes smaller than 2 cm at reefs are suggested for management purposes based on regression equations. This could minimize the amount of movement that occurs when oysters are subjected to boat wakes. The results of this study can help resource managers implement boating policies in Mosquito Lagoon, and contribute greatly to conserving this important ecosystem engineer.

Populations of Crassostrea virginica within Mosquito Lagoon, Florida have recently undergone significant die-offs, which are a subject of major concern. Restoration efforts within Mosquito Lagoon are focusing on reconstructing the three-dimensional reef habitats. Before effective protocols can be established, however, important questions about the sources of juvenile and adult oyster mortality must be answered. Potential causes of Crassostrea virginica mortality in the Indian River Lagoon system include sediment loads, competition, predation, and disease. My research focused on the interactions between oysters and the competitors that may affect the settlement, growth, and survival of Crassostrea virginica. The four objectives of my thesis research were to: 1) identify potential oyster competitors in Mosquito Lagoon, 2) determine if the sessile species recruiting to oyster shells have changed over time, 3) determine how the dominant competitors, barnacles, affect oyster settlement, growth and survival, and 4) determine if oyster or barnacle larvae are better able to settle in increased sediment and flow conditions that are associated with high levels of recreational boating. Lift nets were deployed within Mosquito Lagoon to determine available competing species. I collected species inventory data at six sites to determine the sessile invertebrate species (competitors) present on oyster reefs. Nets were deployed intertidally, just above mean low water, on living oyster reefs. One and a half liters of live and dead oysters were placed within the nets upon deployment. The nets were picked up monthly and surveyed for all fauna. Upon retrieval, all oysters within each net were brought back to the lab where all sessile organisms were immediately identified and returned to the lagoon. This survey began June 2004 and continued for one year. Shells from historic shell middens (up to 15,000 years old) were examined to determine if the sessile species settling on oyster reefs have changed over time. Similar species were found on both shells of historic and extant reefs. One notable exception was the appearance of Balanus amphitrite, an invasive barnacle, on the extant reefs. Balanus amphitrite is thought to have invaded Mosquito Lagoon approximately 100 years ago. This has resulted in a five fold increase in barnacle abundance per oyster shell. Balanus spp. were identified as important potential competitors and thus my research focused on spatial competition between C. virginica and native versus invasive barnacles of the area. Over 300 barnacles, including a native species, Balanus eburneus, and an invasive, Balanus amphitrite, have been counted on a single oyster shell. To determine how Balanus spp. affected settlement, growth, and survivorship of C. virginica, laboratory and field experiments were conducted in which densities of Balanus amphitrite and Balanus eburneus were manipulated. Density treatments included: no barnacles (control), low, medium, and high coverage of barnacles. Laboratory settlement trials with cultured oyster larvae were run in still water and flow (recirculating flume) using all barnacle density treatments. Additionally, all treatments with 7-day oyster spat were deployed in the field to follow oyster spat growth and survivorship. Settlement was counted by microscopy, and growth and survivorship were measured every 3 days for 4 weeks. Settlement of oysters was affected by barnacle presence only in flowing water. Still water trials showed no oyster preference related to any barnacle density or species. The presence of barnacles affected the growth and survivorship of oyster spat. However, there were no species specific differences. Studies suggest that recreational boating activities, especially boat wakes that cause sediment resuspension, may decrease recruitment and this may then provide an advantage to sessile competitors less affected by flow and sediment loads. To address these issues, replicated laboratory trials were run in a laboratory flume to quantify the effects of water motion (0, 5, 10 cm/s) and sediment loads (0, 8, 16 g/ml) on oyster recruitment and the recruitment of an important, relatively new competitor in the system, the barnacle Balanus amphitrite. If B. amphitrite settles in a wider variety of flow rates and sediment conditions, it may have a competitive advantage over the native oyster in this space-limited habitat. I found that high flow and sediment loads reduced larval settlement of C. virginica. Alternatively, settlement of cyprids of B. amphitrite did not differ among treatments. Thus, continuous boat traffic during settlement times should favor recruitment of the invasive barnacle Balanus amphitrite over the native oyster Crassostrea virginica. Determination of the competitive interactions of Crassostrea virginica in Mosquito Lagoon gives us important insights into the ecological conditions necessary for reestablishment of these oyster populations. Crassostrea virginica in Mosquito Lagoon was significantly impacted by barnacles; settlement, growth, and survivorship were all reduced by Balanus spp. This information will help resource managers in planning restoration techniques to minimize oyster and barnacle competitive interactions and increase Crassostrea virgininca success.

The goal of this project was to determine the effects of oyster reef restoration on oyster coverage, wave height dissipation and seagrass recruitment. First, to assess the current versus historical coverage of natural, dead and restored oyster reefs within Mosquito Lagoon, aerial photographs from 2009, provided by Saint Johns River Water Management District, were digitized using ArcGIS software. Live reefs, restored reefs and dead reefs were screen digitized using a reef ‘signature’ in order to estimate the area of each type of reef. The 2009 maps were used as a guide to digitizing the historical aerial photographs (1943, 1951, 1967, 1971, 1984, 1995, 2006). Dead reefs increased both in number and aerial extent during the study period (1943 – 2009), with 2009 having over 10 acres of dead reef coverage. Dead reefs were more likely to be found along major boating channels. Several dead reefs exhibited migration into the mangrove islands located landward of the 1943 footprint, with some dead reefs completely washing up into the shoreline. Restoration of dead reefs added 0.5 acres of live oysters to the Mosquito Lagoon area as of January 2009. Second, I examined how different oyster reef types (natural reef, restored reef, dead reef) reduced wave height. To determine wave height attenuation on each reef type, experiments were conducted in a 9 meter long wave tank using sensors that measured changes in wave height. For each reef type, replicate reefs were created in the wave tank. Shoreline sediment without oysters was used as a control. Using the wave generator, wave heights similar to Mosquito Lagoon boat wakes were created. Restored reefs reduced the incoming wave height by 25% compared to sediment without oysters. iii Lastly, I examined the potential link between oyster reef restoration and recruitment of seagrass fragments. Monthly surveys were performed to quantify the number of seagrass fragments encountering the three oyster reef types: dead reefs, natural reefs and restored reefs. The quantity of seagrass fragments was found to be similar on the three different reef types, but did show a significant trend of seasonality, which corresponds with the growing season of Halodule wrightii. Next, I tested retention of experimentally manipulated seagrass fragments on five natural and five restored reefs. Restored reefs retained seagrass fragments for significantly longer than natural reefs. I also measured seagrass fragment entanglement on each reef type inside the wave tank. I found that seagrass fragments were significantly more likely to become entangled and retained on restored reefs compared to dead and natural reefs. Overall, these metrics are important for determining the success of long-term oyster restoration project in Mosquito Lagoon, Florida. This project has found that oyster restoration is increasing the area of oyster habitat as well as providing important ecosystem services.

As the world continues to experience substantial rates of habitat loss, habitat restoration has become of prime interest to ecologists worldwide. Restoration has shown to be successful in recovering targeted components of certain ecosystems but it is important to achieve a holistic understanding of the resulting ecological impacts it has on communities. To address this, four oyster reefs and three living shorelines were restored during the summer of 2017. These sites, along with four dead oyster reefs, four living oyster reefs, and three undisturbed (control) living shorelines, were sampled before restoration and regularly post-restoration for one year using lift nets. Macroinvertebrates were collected and enumerated in the lab. Diversity indices, community composition, and similarity percentages were then calculated and compared across treatments, time, and treatment-by-time. Live reefs displayed significantly higher species richness and Shannon diversity than restored and dead reefs. Simpson diversity did not differ between live and restored oyster reefs but both were significantly higher than dead reefs. Though not statistically detectable, species richness and Shannon diversity on restored reefs were relatively similar to dead reefs before restoration but became increasingly similar to live reefs over the course of the study. Additionally, analyses revealed significantly different community compositions between live reefs and restored reefs, as well as between live and dead reefs. Living shorelines showed no significant differences in diversity indices but did experience similar seasonal fluctuations in diversity across treatments. Just as with oyster reefs, restored and control living shorelines harbored significantly different communities across time. The findings of this study emphasize the need for dedication to thorough monitoring and multi-metric evaluation of success in restoration efforts. This study and future research will equip resource managers with ways to quantify the effects of restoration that will consider several important ecosystem components.

Aquatic animals, such as fish and shellfish, provide important economic and nutritional benefits for human society. Due to overexploitation of natural fish sources through traditional wild-caught fisheries, aquaculture (generally described as fish farming or culturing) has grown into an economically important industry. A major focus area for the aquaculture field is related to sustainability by ensuring the health and welfare of the aquatic animals. Communities of microorganisms inhabiting the various niches of a given host comprise its microbiome and provide several key health benefits. The microbiome impacts nutrient acquisition, gut homeostasis, protection against pathogens, and immune system modulation. Therefore, much attention has been placed on studying how various culturing conditions and host factors impact the microbiomes of aquatic animals. Here, multiple studies were conducted to elucidate the impacts of various parameters on the microbiomes of rainbow trout, steelhead trout, and Nile tilapia, including dietary supplementation, administration of probiotics and animal age. Though there is a significant correlation between the diet fed to fish and their microbiome communities, small dietary changes such as the inclusion of a dried and lysed yeast product, acting as a protein source alternative to unsustainable fishmeal did not significantly alter the intestinal adherent microbiome of rainbow trout. Moreover, an optimal percentage of yeast replacement that did not negatively impact weight gain for the aquaculture-raised fish was identified, suggesting its efficacy for the industry. Similarly, the intestinal adherent microbiomes of steelhead trout were not significantly altered by diet supplementation with a Bacillus subtilis probiotic. The total microbiome of steelhead trout (mucosa combined with digesta) was instead significantly changed when they were only fed the probiotic additive at an early stage of intestinal development. This change in the microbiome of steelhead trout correlated with a significant increase in weight gain compared to fish only fed the probiotic during later stages of intestinal development. These findings also corroborate previous observations wherein the intestinal microbiome of fish varies during their developmental stages but then stabilizes over time. Determining the core set of bacteria present in fish microbiomes, independent of treatment variables, is another important factor when considering attempts to manipulate the microbiome. To that end, a literature review was conducted in which the phyla Firmicutes, Proteobacteria and, to a lesser extent, Actinobacteria, Bacteroides, and Tenericutes were identified as likely members of the rainbow trout core microbiome. Bacterial families identified as part of the core phyla included Lactobacilliaceae that are commonly used as probiotics and Mycoplasmataceae that lack cell walls. Preventing dysbiosis of the rainbow trout microbiomes will be crucial to ensuring the health of the fish hosts and increasing longevity and profitability of the aquaculture industry. Another important aquaculture-raised species is the Eastern oyster. This animal is critical for the ecological health of the Chesapeake Bay, and it is also an important source of revenue. A significant portion of the revenue flow is the harvest and sale of live oysters for consumption. Unfortunately, consumption of raw or undercooked oysters is the most common route of infection by the human pathogen Vibrio parahaemolyticus (VP) as oysters are a natural reservoir for VP. This bacterium is responsible for a debilitating acute gastroenteritis with potential to cause fatal septicemia. Despite efforts to mitigate infection by this CDC-reportable pathogen, cases continue to increase. The understudied host-microbe relationship between the Eastern oyster and VP has been implicated as a path to research for potential future therapeutics. A novel culturing system for oysters was created using fermentation jars within a BSL-2 ready biosafety cabinet. Using this system, the effect of harvest season was tested against the inoculation efficiency of VP. It was found that higher native Vibrio levels within the oysters were present during the summer compared to the winter. Moreover, addition of the bacteriostatic antibiotic chloramphenicol (Cm) enabled a higher inoculation efficiency by VP during both the summer and winter compared to oysters not exposed to the antibiotic. During the winter, exposure to Cm led to the highest inoculation efficiency (~100%). These findings confirm the importance of the existing microbial communities against exogenous inoculation. Therefore, a year-long study was conducted to investigate the microbiome of oysters during each season. This pan-microbiome study identified a significant impact of harvest season on the microbiome structure. An increased diversity, including higher levels of Cyanobacteriaceae, was observed during the summer. Whereas an increase in Arcobacteriaceae was observed during the winter. Bacteria that persisted throughout the year included Mycoplamataceae and Spirochaeteacae; these families may represent potential members of the Eastern oyster core microbiome. Further work is needed to study the localization patterns of VP within oysters. Such work includes further optimization of immunohistochemistry (IHC) and intracellular colonization assay methods under development here. Collectively, studies of the oyster-microbe interactions will help improve aquaculture methods and identify mitigation targets to reduce VP-related clinical infections. / Doctor of Philosophy / Fish and shellfish provide important economic and nutritional benefits for human society across the globe. Unfortunately, over-fishing of traditional sources of fish and shellfish has led to a reduced supply for world markets, even as the human population increases. Aquaculture, or fish farming, has been around for centuries, but its role in society has significantly increased in the past 50 years. It currently provides about half of fish and other aquatic products on the market today. To better maintain and increase the sustainability and profitability of this industry, more focus is being placed on the health of the fish. The microbiome is the collection of communities of microorganisms, including bacteria, fungi, and archaea, that inhabit various environments including animal hosts. The majority of this dissertation focuses on the impact of factors like diet and age on the microbiomes of aquaculture-raised animals, especially fish. Dietary changes such as the addition of dried yeast-products had a significant impact on fish health but not on the microbiome communities. However, a common probiotic, Bacillus subtilis, did significantly increase not only the growth rate of trout but it also significantly altered the total intestinal microbiome found in the feces and the intestinal mucosal layer. Moreover, it was found that early exposure of the animals to the probiotic had enhanced benefits even though the microbiome appeared to stabilize over time as the fish developed. Maintaining or improving the microbiomes of fish, paying close attention to the microbes that exist as part of a core group of bacteria always present, is vital to ensuring fish health and understanding vertebrate host-microbe relationships. Thus, an analysis of the core microbiome of trout was performed. The final set of projects within this dissertation focused on the relationship between the Eastern oyster, a mollusk native to the Chesapeake Bay, and the bacterial human pathogen Vibrio parahaemolyticus (VP). VP is the leading cause of seafood-borne acute gastroenteritis worldwide, and efforts are needed to mitigate the increasing rate of human infections. Therefore, a simple system using fermentation jars within the laboratory biosafety cabinet was designed to enable safe culture of oysters that were exposed to VP under experimentally controlled conditions. Oysters harvested during the summer naturally harbored higher amounts of native Vibrio organisms in contrast to the winter oysters that harbored much lower levels. A separate microbiome analysis revealed large shifts in the oyster microbiome between summer and winter, although some microbes were continually present. The lower levels of existing Vibrio species detected in winter oysters may have allowed for the higher efficiency of inoculation of winter animals by VP. In fact, these winter animals had Vibrio microbiomes that were completely dominated by the inoculated strain which will enable future work to observe the pattern by which VP localizes, or colonizes, the oysters. Ultimately, these efforts may lead to the development of future disease mitigation strategies against VP.

Cultivated seafood is a potential alternative protein source that can address the rising global food demand with exponentially rising human population growth. Cultivated seafood is made by growing animal cells in vitro using stem cells for edible food, eliminating the need to raise the entire animal. A crucial first step in developing cultivated seafood is creating a well-characterized cell line that can continuously grow and differentiate into desired cell types. Due to difficulties in determining optimal primary cell culture conditions, no continuous cell lines of food-relevant mollusks have been established so far. This study used the adult Eastern Oyster, Crassostrea virginica, as a model bivalve to study the decontamination, cell dissociation, and culture conditions suited for mollusk adductor muscle cells. Oyster adductor (OAD) cells were obtained via tissue explant, mechanical and enzymatic digestion. The cells were routinely monitored using an inverted microscope for phase-contrast and fluorescence imaging. Culture vessels were coated with surface proteins such as fibronectin, laminin, matrigel, and poly-d-lysine to promote cell attachment. The tissue decontamination with Penicillin-Streptomycin (100 µg/mL), Amphotericin B (0.25 µg/ml), and algaecide solution (0.03%) was effective in controlling microbial growth. OAD cells grew best at lower nutrient levels in a one-to-one ratio of Lebovitz L-15 media and artificial seawater. Lower fetal bovine serum levels, 1-5%, provided a high number of cell attachments and consistent growth in combination with 1% adult oyster whole-body or larvae extract. The tissue explant method resulted in the optimal cell dissociation from the three methods, and proceeding cultures had attached cells surviving for up to 10 days. All the plate coatings promoted cell attachment, but fibronectin provided optimal cell attachment of OAD cells. Fibroblast-like, neuron-like, epithelial-like, and rounded cells were observed. Fluorescence cell staining confirmed the presence of cytoskeleton and nuclei in the OAD cell cultures. These advances in primary cell culture methods of OAD cells may be beneficial for establishing mollusk cell lines for cultivated seafood production. / Master of Science in Life Sciences / For sustainable seafood production, alternative sources of seafood proteins are essential in ensuring food security in the future. Cultivated seafood is an alternative protein source to address this rising food demand without the need to raise, farm, or slaughter animals. In developing cultivated seafood, self-renewing stem cells of the animal of interest are grown and made into edible products. A crucial first step in making cultivated seafood is understanding the growth conditions of the primary cells taken from animal tissue. Marine mollusk composes a significant part of seafood consumption, and developing cultured mollusks can address the growing food demand as a seafood alternative. However, there are many gaps in understanding the biological and physiological requirements of mollusk cells. No continuous, self-renewing mollusk cells of food-relevant species have yet been established. This study used the adult Eastern Oyster, Crassostrea virginica, as a model bivalve to study the tissue decontamination, cell dissociation, and culture conditions suited for oyster adductor muscle (OAD) cells. OAD cells were obtained via three cell dissociation methods. Cell growth was routinely monitored using an inverted microscope. Cell-surface proteins such as fibronectin, laminin, matrigel, and poly-d-lysine were used to promote cell attachment. The tissue decontamination was effective with Penicillin-Streptomycin, Amphotericin B, and algaecide. OAD cells grew best at lower nutrient levels in the one-to-one ratio of Lebovitz L-15 media and artificial seawater. Lower fetal bovine serum levels, 1-5%, provided a high number of cell attachments and consistent growth in combination with 1% adult oyster whole-body or larvae extract. Various cell morphologies were observed in the OAD cell cultures. Fluorescence cell staining confirmed the presence of cytoskeleton and nuclei in the OAD cell cultures. These advances in cell culture methods of OAD cells may be beneficial for establishing mollusk cell lines for cultivated seafood production.

Predation may be a key component of the unsuccessful restoration of the Eastern Oyster (Crassostrea virginica), a former keystone species in Chesapeake Bay. Here, I examine the polyclad flatworm Stylochus ellipticus and its potential role as an important predator of C. virginica. Using small-fragment size C. virginica specific DNA primers, oyster DNA was successfully detected in whole organisms homogenates of wild-caught S. ellipticus individuals. Of the 1,575 individuals tested, 68.1% tested positive, thus predation occurred. Predation did not appear to be affected by salinity or temperature; however, season did appear to have an effect on both predation and S. ellipticus abundance (p-value: <0.05). The findings also imply that S. ellipticus are highly mobile, entering the water column to reach hard substrate at various depths, whereas previous studies suggest otherwise. These findings are useful in the planning and management of oyster cultivation and restoration. Furthermore, this study outlines a method of diet study that may be more sensitive than traditional DNA-based techniques.

The (Chilean) flat oyster, Ostrea chilensis, is native to New Zealand and the west coast of South America. It is a commercially important species in New Zealand because of its exquisite taste that attracts premium prices. This thesis describes the first isolation and partial charcterisation of an oyster haemolymph calcium-dependent carbohydrate-binding protein. This protein 'chiletin' was originally isolated from oyster haemolymph by binding to the agarose-galactan matrix of a Sepharose column. Chiletin was predominantly composed of a 24 kilodalton (kDa) band when examined with one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis under non-reducing conditions and a 12 kDa band with reduction of disulphide bonds. The N-terminal sequence of the 24 kDa band was determined to be 'IAGPGWEKYN'. This sequence was not homologous to any known protein. Examination of isolated chiletin with two-dimensional protein analysis gel electrophoresis revealed the presence of three (~12 kDa) subunits ranging in isoelectric point from 5.2 to 6.0. The 24 kDa protein was used to immunise rabbits and a separate antiserum was also raised in rabbits using a synthetic peptide (identical to that above) coupled to keyhole limpet haemocyanin. These antisera were used to confirm the size of the chiletin subunits with Western blots and to examine the elution of chiletin in oyster haemolymph with size exclusion chromatography in phosphate buffered saline (PBS) and 8 M urea. There were four or five different sized conformational aggregates of chiletin present in oyster haemolymph under physiological conditions (PBS). The use of 8 M urea produced two separate aggregates. A major characteristic of lectins is the ability to agglutinate sheep red blood cells and both whole oyster haemolymph and isolated chiletin had this property. Chiletin was identified by immunohistochemistry to be present in a number of tissues. Staining intensity was most consistent in the auricular myocardial cells, followed by the digestive gland epithelium. Chiletin was not induced in haemolymph in response to temperature (30??C) stress or injection of turpentine into the adductor muscle. There have been few immunological studies performed with O. chilensis. The results of the project contribute to what is known about comparative immunology. Greater understanding of how oysters respond to stress and deal with pathogens will ultimately be of benefit to the aquaculture industry.

Oyster reefs are biodiverse communities that provide many ecological and commercial benefits. However, oyster reefs have declined around the world from human activities. Oyster reef restoration programs have begun to limit some of the decline, but the need for determining the success of a program has been problematic. Passive acoustic techniques can use naturally occurring sounds produced by organisms to assess biodiversity. Passive acoustics was utilized to compare the sounds in natural and restored oyster reefs, with special attention on snapping shrimp (Alpheus spp.) snap sounds, in the St. Lucie Estuary, Florida over a one year period. Season, estuary region, habitat and day period had an effect on sound production. Passive acoustic monitoring of snapping shrimp sound production may be a useful non-destructive technique for monitoring the progress of oyster reef restoration projects once further correlations are established between environmental effects and sound production. / by Hilde P. Zenil Becerra. / Thesis (M.S.)--Florida Atlantic University, 2012. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2012. Mode of access: World Wide Web.