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Comparison of American Fisheries Society (AFS) standard fish sampling techniques and environmental DNA for characterizing fish communities in a large reservoir

Recently, methods involving examination of environmental DNA (eDNA) have shown promise for characterizing fish species presence and distribution in waterbodies. We evaluated the use of eDNA for standard fish monitoring surveys in a large reservoir. Specifically, we compared the presence, relative abundance, biomass, and relative percent composition of Largemouth Bass Micropterus salmoides and Gizzard Shad Dorosoma cepedianum measured through eDNA methods and established American Fisheries Society standard sampling methods for Theodore Roosevelt Lake, Arizona. Catches at electrofishing and gillnetting sites were compared with eDNA water samples at sites, within spatial strata, and over the entire reservoir. Gizzard Shad were detected at a higher percentage of sites with eDNA methods than with boat electrofishing in both spring and fall. In contrast, spring and fall gillnetting detected Gizzard Shad at more sites than eDNA. Boat electrofishing and gillnetting detected Largemouth Bass at more sites than eDNA; the exception was fall gillnetting, for which the number of sites of Largemouth Bass detection was equal to that for eDNA. We observed no relationship between relative abundance and biomass of Largemouth Bass and Gizzard Shad measured by established methods and eDNA copies at individual sites or lake sections. Reservoirwide catch composition for Largemouth Bass and Gizzard Shad (numbers and total weight [g] of fish) as determined through a combination of gear types (boat electrofishing plus gillnetting) was similar to the proportion of total eDNA copies from each species in spring and fall field sampling. However, no similarity existed between proportions of fish caught via spring and fall boat electrofishing and the proportion of total eDNA copies from each species. Our study suggests that eDNA field sampling protocols, filtration, DNA extraction, primer design, and DNA sequencing methods need further refinement and testing before incorporation into standard fish sampling surveys.

Arizona↗

Sources and transformations of nitrate from streams draining varying land uses: Evidence from dual isotope analysis

Knowledge of key sources and biogeochemical processes that affect the transport of nitrate (NO 3 - ) in streams can inform watershed management strategies for controlling downstream eutrophication. We applied dual isotope analysis of NO 3 - to determine the dominant sources and processes that affect NO 3 - concentrations in six stream/river watersheds of different land uses. Samples were collected monthly at a range of flow conditions for 15 mo during 2004-05 and analyzed for NO 3 - concentrations, ?? 15 N NO3 , and ?? 18 O NO3 . Samples from two forested watersheds indicated that NO 3 - derived from nitrification was dominant at baseflow. A watershed dominated by suburban land use had three ?? 18 O NO3 values greater than +25???, indicating a large direct contribution of atmospheric NO 3 - transported to the stream during some high flows. Two watersheds with large proportions of agricultural land use had many ?? 15 N NO3 values greater than +9???, suggesting an animal waste source consistent with regional dairy farming practices. These data showed a linear seasonal pattern with a ?? 18 O NO3 :?? 15 N NO3 of 1:2, consistent with seasonally varying denitrification that peaked in late summer to early fall with the warmest temperatures and lowest annual streamflow. The large range of ?? 15 N NO3 values (10???) indicates that NO 3 - supply was likely not limiting the rate of denitrification, consistent with ground water and/or in-stream denitrification. Mixing of two or more distinct sources may have affected the seasonal isotope patterns observed in these two agricultural streams. In a mixed land use watershed of large drainage area, none of the source and process patterns observed in the small streams were evident. These results emphasize that observations at watersheds of a few to a few hundred km 2 may be necessary to adequately quantify the relative roles of various NO 3 - transport and process patterns that contribute to streamflow in large basins. Copyright ?? 2009 by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America. All rights reserved.

Journal of Environmental Quality↗

Potential utility of environmental DNA for early detection of Eurasian watermilfoil ( Myriophyllum spicatum )

Considering the harmful and irreversible consequences of many biological invasions, early detection of an invasive species is an important step toward protecting ecosystems (Sepulveda et al. 2012). Early detection increases the probability that suppression or eradication efforts will be successful because invasive populations are small and localized (Vander Zanden et al. 2010). However, most invasive species are not detected early because current tools have low detection probabilities when target species are rare and the sampling effort required to achieve acceptable detection capabilities with current tools is seldom tractable (Jerde et al. 2011). As a result, many invasive species go undetected until they are abundant and suppression efforts become costly. Novel DNA-based surveillance tools have recently revolutionized early detection abilities using environmental DNA (eDNA) present in the water (Darling and Mahon 2011, Bohmann et al. 2014). In brief, eDNA monitoring enables the identification of organisms from DNA present and collected in water samples. Aquatic and semiaquatic organisms release DNA contained in sloughed, damaged, or partially decomposed tissue and waste products into the water and molecular techniques allow this eDNA in the water column to be identified from simple and easy-tocollect water samples (Darling and Mahon 2011). Despite limited understanding of the production, persistence, and spread of DNA in water (Barnes et al. 2014), eDNA monitoring has been applied not only to invasive species (Jerde et al. 2011), but also to species that are rare, endangered, or highly elusive (Spear et al. 2014). However, most eDNA research and monitoring has focused on detection of invertebrates and vertebrates and less attentionhas been given to developing eDNA techniques for detecting aquatic invasive plants. Eurasian watermilfoil (EWM; Myriophyllum spicatum L.) is an invasive species for which improved early detection would be particularly helpful. Advanced EWM invasions have negative impacts on native biodiversity, recreational boating, fishing, and other types of aquatic tourism (e.g., Eiswerth et al. 2000). On a broader scale, EWM can also be harmful to man-made aquatic infrastructure, such as hydroelectric dams. If an EWM invasion can be detected in an early stage where eradication is still a possibility, many of these negative consequences can be limited or prevented altogether (e.g., Madsen et al. 2002). The purpose of this research was to develop and validate a traditional polymerase chain reaction (PCR) assay for the detection of pure and hybridized EWM DNA using both laboratory and field experiments. We performed a pilot experiment in outdoor tanks to determine the basic functionality and sensitivity of the assay. Following this initial test, we collected field samples from Michigan and Montana lakes with and without known EWM populations. Taken together, our findings suggest that eDNA techniques have potential to be a useful strategy for the early detection of EWM.

Journal of Aquatic Plant Management↗

Relating net nitrogen input in the Mississippi River Basin to nitrate flux in the Lower Mississippi River--A comparison of approaches

A quantitative understanding of the relationship between terrestrial N inputs and riverine N flux can help guide conservation, policy, and adaptive management efforts aimed at preserving or restoring water quality. The objective of this study was to compare recently published approaches for relating terrestrial N inputs to the Mississippi River basin (MRB) with measured nitrate flux in the lower Mississippi River. Nitrogen inputs to and outputs from the MRB (1951 to 1996) were estimated from state-level annual agricultural production statistics and NO y (inorganic oxides of N) deposition estimates for 20 states that comprise 90% of the MRB. A model with water yield and gross N inputs accounted for 85% of the variation in observed annual nitrate flux in the lower Mississippi River, from 1960 to 1998, but tended to underestimate high nitrate flux and overestimate low nitrate flux. A model that used water yield and net anthropogenic nitrogen inputs (NANI) accounted for 95% of the variation in riverine N flux. The NANI approach accounted for N harvested in crops and assumed that crop harvest in excess of the nutritional needs of the humans and livestock in the basin would be exported from the basin. The U.S. White House Committee on Natural Resources and Environment (CENR) developed a more comprehensive N budget that included estimates of ammonia volatilization, denitrification, and exchanges with soil organic matter. The residual N in the CENR budget was weakly and negatively correlated with observed riverine nitrate flux. The CENR estimates of soil N mineralization and immobilization suggested that there were large (2000 kg N ha −1 ) net losses of soil organic N between 1951 and 1996. When the CENR N budget was modified by assuming that soil organic N levels have been relatively constant after 1950, and ammonia volatilization losses are redeposited within the basin, the trend of residual N closely matched temporal variation in NANI and was positively correlated with riverine nitrate flux in the lower Mississippi River. Based on results from applying these three modeling approaches, we conclude that although the NANI approach does not address several processes that influence the N cycle, it appears to focus on the terms that can be estimated with reasonable certainty and that are correlated with riverine N flux.

Journal of Environmental Quality↗

Body mass changes of dabbling and diving ducks wintering in California

Bird body mass is often used as an index of body condition and fluctuates throughout the year in response to environmental conditions and avian life-history events. We examined the body mass of 59,572 ducks representing 13 species (7 dabbling duck species and 6 diving duck species) harvested within the 3 regions of the Central Valley in California, USA (Sacramento Valley, Suisun Marsh, San Joaquin Valley). Data collection occurred in winter during 5 hunting seasons (2014–2015, 2015–2016, 2017–2018, 2018–2019, 2019–2020). For all species, age, and sex classes, the body mass of dabbling ducks varied temporally and was lowest at the end of the hunting season in late January, declining from the beginning of the hunting season in mid-October (northern pintail [ Anas acuta ]: −11.4% to −20.4%; northern shoveler [ Spatula clypeata ]: −3.5% to −17.3%; cinnamon teal [ Anas cyanoptera ]: −5.0% to −17.1%; American wigeon [ Mareca americana ]: −6.9% to −12.2%; American green-winged teal [ Anas carolinensis ]: −5.9% to −11.9%; mallard [ Anas platyrhynchos ]: −1.8% to −9.6%; gadwall [ Mareca strepera ]: −2.2% to −8.4%). As expected, adults (after hatch-year) were heavier than immature (hatch-year) birds (within males: 0.6–8.6%; within females: 0.1–6.6%) and males were heavier than females (within adults: 3.1–38.6%; within immatures: 5.4–33.5%) at the end of the hunting season in all species. Within dabbling duck species, body masses did not differ among regions at the beginning of the hunting season but were heavier in the Sacramento Valley (0.5–14.4%) than other areas by the end of the hunting season. In contrast, body masses of diving ducks did not vary substantially during the hunting season or among regions. Diving ducks demonstrated inconsistent changes in mass from the beginning to the end of the season (lesser scaup [ Aythya affinis ]: 3.4% to 12.3%; canvasback [ Aythya valisineria ]: 5.1% to 6.1%; bufflehead [ Bucephala albeola ]: −3.2% to 1.6%; ring-necked duck [ Aythya collaris ]: −4.9% to 2.6%; common goldeneye [ Bucephala clangula ]: −3.2% to −2.4%; ruddy duck [ Oxyura jamaicensis ]: −16.5% to 5.5%). The substantial temporal and spatial differences in dabbling duck body masses suggest that habitat quality (as measured by caloric value of the available food) or quantity may decline during winter and varies regionally within California's Central Valley.

California↗