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During May 2015, passive acoustic recorders were deployed at eight subtidal oyster reefs within Harris Creek Oyster Sanctuary in Chesapeake Bay, Maryland USA. These sites were selected to represent both restored and unrestored habitats having a range of oyster densities. Throughout the survey, the soundscape within Harris Creek was dominated by the boatwhistle calls of the oyster toadfish, Opsanus tau. A novel, multi-kernel spectral correlation approach was developed to automatically detect these boatwhistle calls using their two lowest harmonic bands. The results provided quantitative information on how call rate and call frequency varied in space and time. Toadfish boatwhistle fundamental frequency ranged from 140 Hz to 260 Hz and was well correlated (r = 0.94) with changes in water temperature, with the fundamental frequency increasing by ~11 Hz for every 1°C increase in temperature. The boatwhistle call rate increased from just a few calls per minute at the start of monitoring on May 7th to ~100 calls/min on May 10th and remained elevated throughout the survey. As male toadfish are known to generate boatwhistles to attract mates, this rapid increase in call rate was interpreted to mark the onset of spring spawning behavior. Call rate was not modulated by water temperature, but showed a consistent diurnal pattern, with a sharp decrease in rate just before sunrise and a peak just after sunset. There was a significant difference in call rate between restored and unrestored reefs, with restored sites having nearly twice the call rate as unrestored sites. This work highlights the benefits of using automated detection techniques that provide quantitative information on species-specific call characteristics and patterns. This type of non-invasive acoustic monitoring provides long-term, semi-continuous information on animal behavior and abundance, and operates effectively in settings that are otherwise difficult to sample.

Enhancingpopulations ofsuspension feedingbivalves,particularlythe easternoyster,Crassostreavirginica, has been proposed as a means of mitigating eutrophication in coastal waters. Review of studies evaluating the effects of C. virginica on nitrogen (N) cycling found that oysters can have effects on water quality that vary by orders of magnitude among sites, seasons, and growing condition (e.g., oyster reefs, aquaculture). Nitrogen contained in phytoplankton consumed by oysters may be returned to the water column, assimilatedintooystertissue and shell,buriedin the sediments,orreturnedtothe atmosphereas dinitrogen gas, primarily via denitrification. Accurately quantifying oyster-related N removal requires detailed knowledge oftheseprimaryfatesofNincoastalwaters.Areviewofexistingdatademonstratedthatthecurrentstateof knowledgeisincompleteinmanyrespects.Nitrogenassimilatedintooystertissueandshellpergramofdry weight was generallysimilaracrosssitesand in oysters growingon reefscomparedto aquaculture.Data on long-term burial of N associated with oyster reefs or aquaculture are lacking. When compared to suitable reference sites, denitrification rates were not consistently enhanced. Depending on environmental and oyster growing conditions, changes in denitrification rates varied by orders of magnitude among studies and did not always occur. Oyster aquaculture rarely enhanced denitrification. Unharvested oyster reefs frequently enhanced denitrification rates. Incorporating oysters into nutrient reduction strategies will require filling gaps in existing data to determine the extent to which relationships between N removal and environmental and/or growing conditions can be generalized.

At a restored reef site and a control site in the Choptank River, Maryland, USA, we partially quantified the effect of oyster reef resto- ration on the removal of nutrients from the water column by determining seasonal fluxes of oxygen (O2), ammonium (NH4 + ), combined nitrate and nitrite (NO2+3), di-nitrogen (N2) and soluble reactive phosphorus (SRP) and by assessing the assimilation of nutrients by macrofauna. Fluxes of O2, NH4 + , NO2+ 3 and SRP at the restored site were enhanced by at least one order of magnitude during all sea- sons. Seasonal denitrification rates at the restored site, measured as flux of N2-N, ranged from 0.3 to 1.6 mmol N2-N m −2 h −1 , with August rates among the highest ever recorded for an aquatic system. In addition to oysters (131 oysters m −2 ; average shell height = 114 mm; age = 2 to 7 yr), the restored reef provided habitat for 24 585 other macrobenthic organisms per square meter compared to 2265 or - ganisms m −2 at the control site. Restoration en - hanced the average standing stock of assimilated nutrients by 95 g N m −2 and 15 g P m −2 . Nitrogen and phosphorus in shells of live oysters and mus- sels accounted for 47 and 48% of total nitrogen and phosphorus standing stocks, respectively. Our results demonstrate that oyster reef restoration can significantly increase denitrification rates and enhance nutrient sequestration via assimilation into bivalve shells.

Oyster reef restoration can significantly increase benthic denitrification rates. Methods applied to measure nutrient fluxes and denitrification from oyster reefs in previous studies include incubations of sediment cores collected adjacent to oyster clumps, benthic chambers filled with intact reef segments that have undergone in situ equilibration and ex situ incubation, and cores with single oysters. However, fluxes of nutrients vary by orders of magnitude among oyster reefs and methods. This study compares two methods of measuring nutrient and metabolic fluxes on restored oyster reefs: incubations including intact segments of oyster reef and incubations containing oyster clumps without underlying sediments. Fluxes of oxygen (O2), dissolved inorganic carbon (DIC), ammonium (NH4+), combined nitrate and nitrite (NO2/3-), di-nitrogen (N2), and soluble reactive phosphorus (SRP) were determined in June and August in Harris Creek, a tributary of the Chesapeake Bay, Maryland, USA. Regression of fluxes measured from clumps alone against those measured from intact reef segments showed significant positive relationships for O2, DIC, NH4+, and SRP (R2 = 0.920, 0.61, 0.26, and 0.52, respectively). Regression of clump fluxes against the oyster tissue biomass indicates significant positive relationships for O2 and NH4+, marginally significant and positive relationships for DIC and N2, and no significant relationship for NO2/3- or SRP. Although these results demonstrate that the incubation of oyster clumps without underlying sediments does not accurately represent biogeochemical fluxes measured from the whole oyster and sediment community, this work supports the need to understand the balance between the metabolism of oysters and local sediments to correctly estimate biogeochemical rates.

We examined the possibility that a nonnative oyster species would provide an ecologically functional equivalent of the native oyster species if introduced into the Chesapeake Bay. Habitat complexity and associated benthic communities of experimental triploid Crassostrea virginica and Crassostrea ariakensis reefs were investigated at 4 sites of varying salinity, tidal regime, water depth, predation intensity, and disease pressure in the Chesapeake Bay region (Maryland and Virginia). Four experimental treatments were established at each site: C. virginica, C. ariakensis, 50:50 of C. virginica and C. ariakensis, and shell only. Abundance, biomass, species richness, evenness, dominance, and diversity of reef-associated fauna were evaluated in relation to habitat location and oyster species. Although habitat complexity varied with location, no differences among complexity were associated with oyster species. Similarly, differences in faunal assemblages were more pronounced between sites than within sites. Our results show functional equivalency between oyster species with respect to habitat at the intertidal site and the low-salinity subtidal location. At subtidal sites of higher salinity, however, the numbers of organisms associated with C. virginica reefs per unit of oyster biomass were significantly greater than the numbers of organisms associated with C. ariakensis reefs. Multivariate analyses of data from subtidal high-salinity sites revealed unique communities associated with C. virginica treatments, whereas mixed-oyster species assemblages were functionally equivalent to monospecific C. ariakensis experimental treatments. Our study represents the first effort to quantify the potential habitat function of C. ariakensis, which has been proposed for an intentional introduction into Chesapeake Bay, and provides evidence of species-specific similarities and differences in reef-associated communities.

In vitro and in vivo infections were conducted to determine if the epizootic hemorrhagic disease (EHD) and bluetongue (BT) viruses would replicate in peripheral blood mononuclear (PBM) cells of white-tailed deer (Odocoileus virginianus). All of the North American EHD and BT viruses (EHD virus serotypes 1 and 2, and BT virus serotypes 2, 10, 11, 13, and 17) replicated in vitro in cultures of white-tailed deer PBM cells. However, this replication appeared to be monocyte-dependent and was not enhanced by lymphocyte blastogenesis induced by the addition of concanavalin A. In white-tailed deer infected with either EHD virus serotype 2 or BT virus serotype 10, virus could be isolated consistently from PBM cells only from post-infection day 4 through 8, although they remained viremic through post-infection day 21. In deer, highest viral titers were associated with the erythrocyte fraction, and in no cases did viral titers detected in the platelet, PBM cell or polymorphonuclear cell fractions approach titers observed in whole blood. In the in vitro infections of white-tailed deer erythrocytes, the EHD and BT viruses were associated with pits in the erythrocyte membrane. This association may be important in the long-term viremia observed in deer.

From 1981 through 1989, serum samples from 855 white-tailed deer (Odocoileus virginianus) from Ossabaw Island, Georgia (USA), were tested for antibodies to bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). During this period, prevalence of precipitating antibodies to BTV and EHDV as determined by agar gel immunodiffusion (AGID) tests decreased from 74% to 3% and from 34% to 1%, respectively. Antibodies were detected in serum samples from 0.5-yr-old deer only during 1981, 1982, and 1983, and with few exceptions, positive serological results after 1983 were restricted to older age classes. A decrease in prevalence of precipitating antibodies to BTV and EHDV in age classes exposed during 1981 indicates that AGID results from white-tailed deer populations underestimate the extent of previous exposure to these viruses. Serum neutralization test results from AGID-positive deer indicated that BTV 11 was the principal serotype responsible for infections during 1981. Since 1983, this serotype has been replaced by BTV 13; however, there has been a low level of transmission within the herd. Infection with EHDV 2 appeared most prevalent during 1982; as with BTV 13, there has been limited transmission in this high density deer population since 1983.