The decline of many freshwater mussel populations in the United States has brought about the need for facilities in which mussels can be held for purposes of relocation, research, and propagation. The Aquatic Wildlife Conservation Center (AWCC) of the Virginia Department of Game and Inland Fisheries (VDGIF) serves as a freshwater mussel conservation facility in southwest Virginia. The goals of this study were: (1) to determine whether adult freshwater mussels could maintain energy reserves at AWCC (2) to determine whether adults could produce mature gametes at AWCC and (3) to establish suitable rearing conditions for juvenile mussels at the AWCC. In fall 2002, four species of mussels, Villosa iris, V. vanuxemensis, Amblema plicata, and Pleurobema oviforme, served as surrogates for endangered species and were relocated to the AWCC. Three energy reserves (glycogen, protein, and lipid) were measured seasonally (fall 2002 to summer 2004) from mantle tissue and compared between AWCC specimens and those from their wild source populations. The gametogenic stage of each species was also compared to determine whether gametogenesis was occurring in captivity. In summer 2003, the first of two juvenile experiments tested the effects of three rates of water flow (1 L/min, 3 L/min, and 7 L/min) on the survival and growth of V. iris and Epioblasma capsaeformis reared in flow-through troughs. In summer 2004, round flow-through tanks were used to assess the effects of three sizes of substrate (fine sediment, fine sand, and coarse sand) and sampling frequency on the survival and growth of V. iris. Gut content analyses also were conducted at the end of each experiment to determine which algal species were being consumed. Overall survival rates were as follows: A. plicata, 100 %; V. vanuxemensis, 86 %; V. iris, 79 %; P. oviforme (2002 collection), 53 %; and P. oviforme (2003 collection), 50 %. All energy reserves varied among seasons, but every species except P. oviforme (2003 collection) had levels higher than those in source populations at the end of this experiment. Glycogen appeared to be the best indicator of condition in these species, with protein also being important in the 2003 collection of P. oviforme. Mature gametes were found in all four captive species in 2003 and 2004, with lipids appearing to fuel gametogenesis. Additionally, gametogenesis was occurring earlier in captive long-term brooders than in the wild, possibly due to warmer water temperatures at AWCC. The first juvenile experiment resulted in 15 % overall survival, with 1 L/min having the greatest survival (18 %), and the 3 L/min having the greatest growth (656 μm). In the second experiment, dishes left unsampled had significantly greater survival (40 %) (P<0.05) of juveniles than those which were sampled (27 %). The unsampled fine sand treatment had significantly greater survival than the other two unsampled treatments (52 %) (P<0.001). Sampled juveniles in fine sediment had the greatest growth (887 μm). Also, juveniles from Experiment 1 were consuming primarily Navicula, with Coelastrum and Chlorella consumed in greatest abundance in Experiment 2. Results indicate that most adult mussels maintained energy reserves and produced mature gametes, and that juveniles of V. iris had good survival and growth. Only P. oviforme had survival rates lower than expected and did not appear to maintain condition at AWCC. Based on results of the species tested, environmental conditions at AWCC appear suitable for the survival of most adult and juvenile freshwater mussels. / Master of Science

Freshwater mussel propagation techniques were tested at the Virginia Department of Game and Inland Fisheries Aquatic Wildlife Conservation Center through a series of three experiments. Experiment 1 tested the suitability of a pond and raceway for rearing juvenile oystermussels (Epioblasma capsaeformis) and wavyrayed lampmussel (Lampsilis fasciola). This experiment was prematurely terminated due to predation on mussels by fathead minnows (Pimephales promelas). Experiment 2 evaluated growth and survival of juvenile rainbow mussels in outdoor troughs and indoor aquaria. There was no significant difference in survival or growth between the two systems. Experiment 3 used troughs similar to those in Experiment 2 to rear E. capsaeformis and L. fasciola under two silt regimes. Survival for Experiment 3 was significantly greater for L. fasciola. The comparison between silt regimes indicated that individuals in the high-silt treatment had better survival than those in the low-silt treatment. The difference between these 2 treatments may be a reflection of increased escapement in the low-silt treatment, which may have resulted from more frequent disturbance during sampling. Growth of L. fasciola was significantly greater than E. capsaeformis, and was greater in the low-silt treatment. A habitat survey of the Clinch River, Virginia was conducted from Blackford, Clinch River Kilometer (CRK) 478 to the Tennessee border, CRK 325. Physical characteristics identified in the survey were combined with water quality and impact source data to develop a habitat suitability index for freshwater mussels within this study reach. Model parameters were indexed and weighted to give a final suitability ranking. Habitat units having the highest overall ranking included: Nash Ford (CRK 449), Artrip (CRK 442), several riffles and runs below Carterton (CRK 417), upstream of Mill Island (CRK 389.5), and Pendleton Island (CRK 365), and Speers Ferry (CRK 333-325). Potential locations for habitat restoration projects and additional monitoring were also identified. / Master of Science

Glochidia (larvae) of freshwater mussels are obligate parasites which attach to and become encysted in the gills or fins of host fish species. The immune responses of the host fish to the parasite affects the susceptibility of the fish to glochidia of different mussels. The immune response provides an opportunity to identify which fish species are hosts. The number and variety of mussels in rivers and lakes has sharply declined since the last century due to various anthropogenic factors, and some mussels species are facing extinction. It is an urgent task to preserve these vanishing mussels, or extinction will be inevitable. An attempt was made to develop an assay, using the immunological response to glochidia, to screen fish species for appropriate hosts. This would facilitate the production and rearing of juveniles. In order to design these assays, reagents such as anti-immunoglobulins which can react with antibodies from many different fish species have to be developed. This work was carried out to develop such reagents. Host and non-host fish were immunized with killed bacteria (Brucella abortus) to study their humoral immune response to an antigen. All fish were able to respond well, as measured by agglutination and Western Blot assays. Antibodies produced by the Brucella injections were used to stimulate anti-fish immunoglobulins in goats, and the antisera were tested for their ability to recognize immunoglobulins from different host fish species. The specificities of these reactions were compared to the reactivity of Protein A. Goat antisera were able to cross-react with different fish antibodies, but it was found that Protein A was a more suitable reagent. Protein A is seemingly suitable to identify the host-fish species and could be used as a reagent for the serological diagnosis of various fish diseases. / Master of Science