Development and Integration of Quantitative Real-Time PCR Methods for Detection of Mitochondrial DNA and Methanobrevibacter smithii nifH Gene as Novel Microbial Source Tracking Tools

A novel microbial source tracking (MST) method based on the detection of human and non-human markers was developed and applied to track the origin of fecal pollution in water systems. Mitochondrial DNA sequences were used to develop new quantitative real-time polymerase chain reaction (qPCR) assays for dog, poultry, and gull. The targets were included as part of a toolbox including human, cow, pig, and sheep assays. A primer and probe set for the detection of the human-specific nifH gene of Methanobrevibacter smithii was also designed as an indicator of human fecal contamination. The assays were tested for specificity and applied to fecal-spiked surface waters and environmental samples collected from two river catchments impacted by sources of human and non-human fecal contamination. The MST methods described were applicable to both spiked waters and environmental samples, and using the two approaches the origin of fecal pollution could be successfully determined in mixed source fecally polluted waters.     

Comparison of LLW Disposal Facilities at Major Department of Energy Sites

Near surface disposal facility design and management are examined and compared using a systems approach that defines facility performance as a function of three components (or subsystems): the disposal facility design (cover systems and bottom liners); the properties of the waste (waste composition, waste form and waste package); and the site‐specific environmental features (climate, geology, and hydrology). We report an evaluation of five DOE near surface disposal facility case studies, selected to provide a “representative” sample that included disposal sites with a range of waste and environmental characteristics across the DOE. The facilities selected were the Savannah River E‐Area Engineered Trenches, Hanford Integrated Disposal Facility, Idaho Radioactive Waste Management Complex, Oak Ridge Environmental Management Waste Management Facility, and Nevada National Security Site Area 5. ©2015 Wiley Periodicals, Inc.     

Sexual conflict over fertilizations: female bluethroats escape male paternity guards

Extra-pair copulations create a potential for sexual conflict in pair-bonding birds. Here we report an experimental study of the bluethroat, Luscinia s. svecica, in which the throat ornament of males was blackened with Nyanzol D in order to reduce their sexual attractiveness and thus increase the sexual conflict over fertilizations. In an earlier study, we showed that males blackened before pairing had a lower success in attracting social mates than controls, whereas males blackened after pairing guarded their mates more intensely and sang less than controls. Here we add behavioural data from one more year on males blackened after pairing and corroborate our previous finding that the manipulation caused males to guard their mates more intensely and advertise less for additional mates. Blackened males did not suffer more intrusions from neighbouring males than did controls. Paternity analyses of the combined data set, using multilocus DNA fingerprinting and microsatellite typing, revealed that blackened males lost significantly more paternity than controls. There was also a tendency for blackened males to show a lower success in achieving extra-pair fertilizations. These results indicate that females have the upper hand in the sexual conflict over fertilizations, as females paired with unattractive males can achieve more extra-pair paternity despite the greater constraint posed by the intensified mate guarding. Still, within the blackened group, there were some indications that males guarding more intensely and singing less had higher paternity than males guarding less and singing more, suggesting a marginal positive effect of guarding for unattractive males. Male mate guarding must nevertheless be considered a best-of-a-bad-job strategy in this species. Received: 4 December 1997 / Accepted after revision: 14 June 1998     

Microbial community dynamics in uranium contaminated subsurface sediments under biostimulated conditions with high nitrate and nickel pressure

BACKGROUND, AIM, AND SCOPE: The subsurface at the Oak Ridge Field Research Center represents an extreme and diverse geochemical environment that places different stresses on the endogenous microbial communities, including low pH, elevated nitrate concentrations, and the occurrence of heavy metals and radionuclides, including hexavalent uranium [U(VI)]. The in situ immobilization of U(VI) in the aquifer can be achieved through microbial reduction to relatively insoluble U(IV). However, a high redox potential due to the presence of nitrate and the toxicity of heavy metals will impede this process. Our aim is to test biostimulation of the endogenous microbial communities to improve nitrate reduction and subsequent U(VI) reduction under conditions of elevated heavy metals. MATERIALS AND METHODS: Column experiments were used to test the possibility of using biostimulation via the addition of ethanol as a carbon source to improve nitrate reduction in the presence of elevated aqueous nickel. We subsequently analyzed the composition of the microbial communities that became established and their potential for U(VI) reduction and its in situ immobilization. RESULTS: Phylogenetic analysis revealed that the microbial population changed from heavy metal sensitive members of the actinobacteria, alpha- and gamma-proteobacteria to a community dominated by heavy metal resistant (nickel, cadmium, zinc, and cobalt resistant), nitrate reducing beta- and gamma-proteobacteria, and sulfate reducing Clostridiaceae. Coincidentally, synchrotron X-ray absorption spectroscopy analyses indicated that the resulting redox conditions favored U(VI) reduction transformation to insoluble U(IV) species associated with soil minerals and biomass. DISCUSSION: This study shows that the necessary genetic information to adapt to the implemented nickel stress resides in the endogenous microbial population present at the Oak Ridge FRC site, which changed from a community generally found under oligotrophic conditions to a community able to withstand the stress imposed by heavy metals, while efficiently reducing nitrate as electron donor. Once nitrate was reduced efficient reduction and in situ immobilization of uranium was observed. CONCLUSIONS: This study provides evidence that stimulating the metabolism of the endogenous bacterial population at the Oak Ridge FRC site by adding ethanol, a suitable carbon source, results in efficient nitrate reduction under conditions of elevated nickel, and a decrease of the redox potential such that sulfate and iron reducing bacteria are able to thrive and create conditions favorable for the reduction and in situ immobilization of uranium. Since we have found that the remediation potential resides within the endogenous microbial community, we believe it will be feasible to conduct field tests. RECOMMENDATIONS AND PERSPECTIVES: Biostimulation of endogenous bacteria provides an efficient tool for the successful in situ remediation of mixed-waste sites, particularly those co-contaminated with heavy metals, nitrate and radionuclides, as found in the United States and other countries as environmental legacies of the nuclear age.     

地下水铀污染的原位微生物还原与固定:在美国能源部田纳西橡树岭放射物污染现场的试验

总结了美国斯坦福大学和橡树岭国家实验室等在美国能源部田纳西州橡树岭综合试验基地进行的铀污染原位微生物修复阶段性试验结果.本试验利用微生物以乙醇为电子供体还原地下水和沉积物中的六价铀为不溶解的四价铀,使之原位固定化.随后通过加入溶解氧和硝酸盐来试验微生物还原后的地下水层中还原固定态铀的稳定性.通过预处理和长期间隔注入乙醇...     

Use of monitoring data to support conservation management and policy decisions in Micronesia

Adaptive management implies a continuous knowledge‐based decision‐making process in conservation. Yet, the coupling of scientific monitoring and management frameworks remains rare in practice because formal and informal communication pathways are lacking. We examined 4 cases in Micronesia where conservation practitioners are using new knowledge in the form of monitoring data to advance marine conservation. These cases were drawn from projects in Micronesia Challenge jurisdictions that received funding for coupled monitoring‐to‐management frameworks and encompassed all segments of adaptive management. Monitoring in Helen Reef, Republic of Palau, was catalyzed by coral bleaching and revealed evidence of overfishing that led to increased enforcement and outreach. In Nimpal Channel, Yap, Federated States of Micronesia (FSM), monitoring the recovery of marine food resources after customary restrictions were put in place led to new, more effective enforcement approaches. Monitoring in Laolao Bay, Saipan, Commonwealth of the Northern Mariana Islands, was catalyzed by observable sediment loads from poor land‐use practices and resulted in actions that reduced land‐based threats, particularly littering and illegal burning, and revealed additional threats from overfishing. Pohnpei (FSM) began monitoring after observed declines in grouper spawning aggregations. This data led to adjusting marine conservation area boundaries and implementing market‐based size class restrictions. Two themes emerged from these cases. First, in each case monitoring was conducted in a manner relevant to the social and ecological systems and integrated into the decision‐making process. Second, conservation practitioners and scientists in these cases integrated culturally appropriate stakeholder engagement throughout all phases of the adaptive management cycle. More broadly, our study suggests, when describing adaptive management, providing more details on how monitoring and management activities are linked at similar spatial scales and across similar time frames can enhance the application of knowledge.     

Parental investment decision rules: a test in bluegill sunfish

Summary We review parental investment decision theory and provide an experimental test of the decision rule used by male bluegill sunfish (Lepomis macrochirus) in allocating parental investment to their young. The alternative decision rules tested are: (1) invest according to brood size (number) only; (2) invest according to past investment only; (3) invest according to both brood size and past investment; and (4) invest according to neither brood size nor past investment. By manipulating brood size independently of a male's cumulative investment in the brood, and by measuring each male's defensive behavior against a model predator, we found that male bluegill invest according to both brood size and past investment. This result is consistent with recent theory that past parental investment devalues adult future reproductive value, and that animals should therefore invest according to the value of their brood relative to that of their own expected future reproduction.     

Proactive environmental programs: Defining customers and measuring their expectations at apple computer

Resources to develop proactive environmental programs compete directly with resources dedicated to the fundamental business of the company. Therefore, environmental managers need to prioritize environmental programs using the same criteria as the company as a whole; for many companies, this means measuring how well such programs meet and exceed customer expectations. We must define our customers, measure their expectations, analyze the results, develop programs in response, and finally measure the results of the program implementation. Although Apple has not completed this cycle, our customer definition and measurement systems are taking shape. This article explains how we define our customers, and how we measure their expectations through direct and indirect data collection mechanisms. It also outlines measurement results and our evolving program focus in response to those results.     

Visual fields in Flamingos: chick-feeding versus filter-feeding

In birds, the position and extent of the region of binocular vision appears to be determined by feeding ecology. Of prime importance is the degree to which vision is used for the precise control of bill position when pecking or lunging at prey. In birds that do not require such precision (probe and filter-feeders), the bill falls outside the binocular field, which extends above and behind the head, thus providing comprehensive visual coverage. Flamingos Phoenicopteridae are highly specialised filter-feeders. They employ a unique technique that does not require accurate bill positioning in which the inverted head is placed between the feet. Feeding flamingos often walk forwards with the head pointing “backwards”. Here we show that in Lesser Flamingos Phoeniconaias minor visual fields are in fact the same as those of birds that feed by precision pecking and that feeding flamingos are blind in the direction of their walking. We suggest that this is due to the requirement for accurate bill placement when flamingos feed their chicks with “crop-milk”, and possibly when building their nest. We propose that chick-feeding may be the ultimate determinant of visual field topography in birds, not feeding ecology.