Propagation and culture has been accepted as an approved aquaculture method for resource managers to enhance and recover freshwater mussel populations. Although juveniles can be produced readily for many mussel species, achieving high growth and survival in the laboratory remains difficult. The goal of my project was to improve growth and survival of juvenile mussels by comparing diets, algae concentration, and culture systems. The first objective determined a suitable diet for feeding juvenile northern riffleshell, Epioblasma torulosa rangiana, a species listed as federally endangered. Three algal diets were evaluated to determine differences in growth and survival of juveniles of E. t. rangiana. After 60 d, mean survival on Phytofeast, Shellfish Diet, and WSSNFH mix were 75.1 (95% CI: 72.2 to 78.0), 78.9 (95% CI: 74.5 to 83.2), and 85.0 (95% CI: 80.6 to 89.3) percent, respectively. WSSNFH mix had the highest survival which was significantly different from Phytofeast (p=0.01).  Mean shell lengths were 2.37 mm (95% CI: 2.27 to 2.47), 2.62 mm (95% CI: 2.52 to 2.72), and 3.11 mm (95% CI: 3.01 to 3.22), respectively.  Juvenile length on all three diet treatments was significantly different from each other (p<0.0001), with the WSSNFH mix exhibiting the highest growth, and Phytofeast with the lowest growth. My second objective evaluated the effect of algal concentration (cells mL-1) on growth and survival of juveniles of E. t. rangiana and mucket, Actinonaias  ligamentina. After 60 d, mean survival of E. t. rangiana for the low (30,000 cells mL-1), medium (80,000 cells mL-1), and high (140,000 cells mL⁻¹) algal concentrations were 39.1 (95% CI: 30.7 to 47.4), 20.7 (95% CI: 12.8 to 28.6), and 12.7 (95% CI: 4.82 to 20.5) percent, respectively (p<0.01). Mean shell lengths were 1.58 mm (95% CI: 1.49 to 1.66), 1.30 mm (95% CI: 1.19 to 1.40), and 1.01 mm (95% CI: 0.936 to 1.08), respectively (p<0.0001).  Mean survival of A. ligamentina for the low, medium, and high algae concentrations were 46.8 (95% CI: 35.2 to 58.4), 24.6 (95% CI: 15.1 to 34.0), and 10.7 (95% CI: 5.45 to 15.9) percent, respectively (p<0.01). Significant differences were observed between the low feed concentration versus the medium and high feed concentrations. Mean shell lengths for the low, medium, and high concentrations were 1.15 mm (95% CI: 1.08 to 1.22), 0.994 mm (95% CI: 0.930 to 1.06), and 0.833 mm (95% CI: 0.770 to 0.896), respectively. All concentrations were significantly different, and the low concentration had the highest mean shell length (p<0.0001). The third objective compared the performance of three recirculating aquaculture systems for rearing juvenile mussels >5 mm. Mean incremental length of juveniles of E. t. rangiana at 60 d in Pans, Buckets, and Upwellers was 1.19 mm (95% CI: 0.746 to 1.62), 1.05 mm (95% CI: 0.608 to 1.49), and 2.07 mm (95% CI: 1.63 to 2.51), respectively. Incremental lengths were significantly higher in the Upwellers (p=0.03). The mean lengths for Bucket and Pan systems were not significantly different from each other (p=0.54).  Percent survival of juveniles for the Pans, Buckets, and Upwellers were 91.7 (95% CI: 87.4 to 96.0), 90.0 (95% CI: 80.6 to 99.4), and 100 (95% CI: 100 to 100), respectively. Survival in the Upwellers was significantly higher than in the Buckets (p=0.018). Survival of juveniles in the Pan system and Upwellers were not significantly different from each other (p=0.05).  Mean growth for A. ligamentina was 1.96 mm (95% CI: 1.03 to 2.9), 0.88 mm (95% CI: 0.048 to 1.80), and 2.46 mm (95% CI: 1.537 to 3.38), respectively (p=0.07). Mean percent survival of juveniles of Actinonaias ligamentina in the Pans, Buckets, and Upwellers were 100 (95% CI: 100 to 100), 86.7 (95% CI: 74.0 to 99.4), and 100 (95% CI: 100 to 100), respectively.   Survival of A. ligamentina in the Upwellers was significantly higher than in Buckets (p<0.0001).  Juvenile survival in the Pan system and Upwellers was not significantly different (p=0.998). Results indicate that the Upweller culture system supported the highest growth and survival in culturing E. t. rangiana and A. ligamentina. / Master of Science

This study deals with the reproductive ecology and larval development of Mytilus californianus. Mytilus californianus is the common mussel found along the more exposed west coast of North America from the Aleutian Islands south Isla Socorro, Mexico (Soot-Ryen, 1955) and was chosen because it is an important species in rocky intertidal communities (being the most abundant if not the most conspicuous) and forms the basis for diverse associations of numerous other species. The reproductive cycle, larval development to settlement, comparisons of reproduction in high and low populations and field observations on larval settlement are described.

Seasonality estimates based on archaeological shellfish remains have been an important component of settlement pattern reconstruction. Investigations of this nature allow researchers to place prehistoric people on the landscape at points in space at different times of the year. Many of the previous seasonality studies, however, have focused on marine species from coastal sites, with little attention given to freshwater locales, especially ones in the Mississippi Delta. To address that disparity, this study examines freshwater mussel “season of capture” via analysis of stable oxygen isotope ratios in specimens recovered from two Late Woodland sites located along the Yazoo River, Mississippi. As freshwater mussel shells are composed of aragonite, a metastable form of calcium carbonate (CaCO3), they can suffer greatly from the impact of meteoric diagenesis. Because of this, samples must be evaluated for diagenesis prior to any geochemical analysis taking place. Archaeological shell samples were examined via thin-section petrography and scanning electron microscopy (SEM). Visual analysis indicated pristine aragonite microstructure and crystallography in all archaeological shell samples, and confirmed their suitability for isotope analysis. Vetted shells were then micromilled across accretionary growth bands, and analyzed for their oxygen isotope signatures. Isotope profiles were then interpreted for their individual “season of capture”, and oscillation patterns for 22 shell specimens indicated mussels were being collected in all four seasons. These data support the view that at least some portion of the human population at both sites engaged in shellfishing activities year-round, indicating sedentary populations at both locales. The shell assemblages were also investigated for the purpose of informing modern conservation efforts (i.e., “applied zooarchaeology”). Nearly 24,000 valves were analyzed taxonomically, yielding the presence of 37 species, of which 24 represented new river records for the Yazoo River. These data provide a valuable historical perspective, cataloging communities as they existed prior to extensive modern impacts, thus representing an ecological baseline to be compared with modern populations. Though modern data are extremely limited for the river, the study revealed it once supported a diverse mussel community containing numerous species currently considered rare, endangered, or extinct in Mississippi.

Predicting the locations of new biological invasions has become a high priority for biologists as well as trying to predict if newly introduced species will become damaging to native ecosystems. Reefs of the eastern oyster Crassostrea virginica in Mosquito Lagoon, Florida have been highly disturbed in recent years resulting in dead reefs (piles of dead, disarticulated shells) some of which have been restored. I conducted oyster reef surveys for non-native invertebrates to determine if disturbance on these oyster reefs might assist invasion by two species, Mytella charruana and Perna viridis, recently introduced to the southeastern coast of the United States. Next, I investigated if M. charruana's temperature and aerial exposure tolerance limits may allow for it to establish permanently on intertidal oyster reefs. Temperature and aerial exposure tolerance experiments were conducted and oyster reef temperatures were collected. Oyster reef surveys could not predict if reef disturbance is assisting in the invasion process because only two non-native individuals (P.viridis) were found, one on a restored reef and one on a natural (reference) reef. Tolerance experiments showed that some Mytella charruana survived even after 7 days of 8??C temperatures if the mussels are exposed to air for 4 hours or less per day. Mytella charruana had near 0% survival after 4 hours of 44??C. However, only disturbed reefs reached this temperature in the field. It is likely that M. charruana could survive in the low intertidal zone on restored or reference reefs. This information is important for understanding the introduction of M. charruana in Mosquito Lagoon and also provides a data set of temperature tolerances for better understanding of whether the species might be able to invade other areas.

In 1998, a tanker truck overturned on U.S. Route 460 in Tazewell County, Virginia, releasing approximately 5110 L of Octocure 554-revised, a rubber accelerant, into an unnamed tributary approximately 162 m from its confluence with the Clinch River. The spill killed nearly all aquatic life in the river, including three species of federally endangered mussels, for 11 km downstream. The restoration plan developed after the spill included community outreach to complement the physical, on-site restoration effort. Although numerous studies have been made of the aquatic resources in the upper Clinch River, Tazewell County, Virginia, there had been no prior assessment of the local residents' knowledge, attitudes, and opinions about the natural resources in this biodiversity hotspot. I surveyed residents of Tazewell County, Virginia to assess their baseline knowledge about the upper Clinch River watershed, endangered mussels, aquatic conservation, and water quality issues. The survey response rate was 42%. Survey respondents were aware that mussels are present in the Clinch River watershed, but were not aware of their imperiled status or the ecosystem services that mussels perform. On average, respondents expressed stronger moralistic, naturalistic, and ecologistic than negativistic and utilitarian attitude orientations toward endangered freshwater mussels. Respondents considered water quality important and expressed support for mussel restoration. However, respondents indicated that the presence of mussel populations might negatively impact economic development of the county. Local community leaders and conservation educators, when interviewed to explore their opinions on conservation of aquatic resources, placed high value upon the Clinch River's water quality. They also have sought information regarding water quality and the Clinch River from local organizations and agencies and/or have developed a relationship with them as a result of their long-term presence in the community. The majority of respondents believed that human impacts contribute to species decline, but human impacts may not be the primary contributor to such decline. Respondents indicated that governmental and nongovernmental conservation agencies and organizations can assist localities by providing additional funding opportunities, seminars, and training sessions. Tazewell County conservation educators focus broadly on water resources of the county. Conservation educators wised that adults and teenagers displayed greater interest in conservation issues and that local governments were involved more actively in conservation. Overwhelmingly, educators believed that forming and fostering partnerships is the most effective way to inform audiences about conservation. Barriers to conservation education faced by educators include lack of funding, audience apathy, and/or audience negativity. Nearly all respondents indicated that balance between conservation and development currently exists or that achieving such a balance should be a goal of the local government. Survey and interview results were used to develop specific outreach recommendations to generate community support for mussel restoration and aquatic conservation in Tazewell County. Recommendations for the Certus Spill Outreach Plan include: frame mussel outreach messages in a broader aquatic ecosystem context in order to emphasize the human connection to aquatic ecosystems, communicate positive messages about conservation with decision-makers, regularly communicate positive messages about conservation in the local press, and partner with the local school system and agencies that have established positive images in the towns and county. / Master of Science

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