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.     

Projecting Large-Scale Area Changes in Land Use and Land Cover for Terrestrial Carbon Analyses

One of the largest changes in US forest type areas over the last half-century has involved pine types in the South. The area of planted pine has increased more than 10-fold since 1950, mostly on private lands. Private landowners have responded to market incentives and government programs, including subsidized afforestation on marginal agricultural land. Timber harvest is a crucial disturbance affecting planted pine area, as other forest types are converted to planted pine after harvest. Conversely, however, many harvested pine plantations revert to other forest types, mainly due to passive regeneration behavior on nonindustrial private timberlands. We model land use and land cover changes as a basis for projecting future changes in planted pine area, to aid policy analysts concerned with mitigation activities for global climate change. Projections are prepared in two stages. Projected land use changes include deforestation due to pressures to develop rural land as the human population expands, which is a larger area than that converted from other rural lands (e.g., agriculture) to forestry. In the second stage, transitions among forest types are projected on land allocated to forestry. We consider reforestation, influences of timber harvest, and natural succession and disturbance processes. Baseline projections indicate a net increase of about 5.6 million ha in planted pine area in the South over the next 50 years, with a notable increase in sequestered carbon. Additional opportunities to expand pine plantation area warrant study of landowner behavior to aid in designing more effective incentives for inducing land use and land cover changes to help mitigate climate change and attain other goals.     

Stochastic modelling of landfill processes incorporating waste heterogeneity and data uncertainty

A landfill is a very complex heterogeneous environment and as such it presents many modelling challenges. Attempts to develop models that reproduce these complexities generally involve the use of large numbers of spatially dependent parameters that cannot be properly characterised in the face of data uncertainty. An alternative method is presented, which couples a simplified microbial degradation model with a stochastic hydrological and contaminant transport model. This provides a framework for incorporating the complex effects of spatial heterogeneity within the landfill in a simplified manner, along with other key variables. A methodology for handling data uncertainty is also integrated into the model structure. Illustrative examples of the model's output are presented to demonstrate effects of data uncertainty on leachate composition and gas volume prediction.     

Neonatal form of the hyperornithinaemia,hyperammonaemia, and homocitrullinuria (HHH) syndrome and prenatal diagnosis

We describe here the first case of neonatal death due to the hyperornithinaemia, hyperammonaemia, and homocitrullinuria (HHH) syndrome and the first prenatal diagnosis of this disorder in a subsequent pregnancy in this family.     

Screening maternal serum alpha-fetoprotein levels and human parvovirus antibodies

The association between gestational infection with human parvovirus (B19) and fetal loss has increased interest in this virus and demand for diagnostic testing. However, serological assays for B19 are not yet widely available. Maternal serum alpha-fetoprotein (MSAFP) testing is commonly used during the second trimester to screen for various fetal defects. We attempted to determine whether an elevated level of MSAFP would be an appropriate indication for B19-specific tests. Over a 26-month period, MSAFP tests were performed at Michigan State University for 21 392 women. Sera remaining after that testing were stored frozen. Of these, 22 case samples—from women with MSAFP levels greater than 3·0 multiples of the median (MOM) and pregnancies that ended in fetal loss—and 44 matched control samples—from women with MSAFP levels greater than 0·4 and less than 2·2 MOM and live births at term—were tested for B19 antibodies. None of the 66 samples was IgM positive, while 33 (50 per cent) were IgG positive. The presence of IgG was not significantly associated with case or control status (matched odds ratio=0·77, 95 per cent confidence interval 0·28–2·11). These findings are consistent with other studies indicating prior infection in approximately half of adults and suggest that elevated screening MSAFP levels, in the absence of other evidence of B19 infection, should not prompt B19-specific testing.     

Recreational scuba diving in Caribbean marine protected areas: do the users pay?

There are more than 200 marine protected areas (MPAs) in the Caribbean and Central America that contain coral reefs and are therefore theoretically attractive to scuba divers. One fifth of dive operators in 30 countries were surveyed for their use of MPAs: the majority are located within 20 km of at least one MPA and 46% conduct at least 80% of their diving within a MPA. An estimated 15 million dives take place outside of Florida each year, half of these occurring inside MPAs. Only 25% of MPAs containing coral reefs charge divers an entry or user fee, which is most usually USD 2-3 levied per dive or per diver. The revenue generated by these fees is estimated at USD 1-2 million annually, but the potential for generating income has not been fully realized. A significant contribution to the cost of regional conservation could be achieved if higher fees were applied more widely than at present.     

The role of asteroid predators in the organization of a sessile community on pier pilings

The diets of 4 species of asteroids were observed for `9 months on the piles of a pier at Rapid Bay, South Australia.Coscinasterias calamaria (Gray) fed on molluscs, crustaceans and moribund items.Patiriella brevispina H.L. Clark fed on compound ascidians, sponges and moribund items.Tosia australis Gray andPetricia vernicina (Lamarck) ate mainly sponges, bryozoans and compound ascidians. A short-term (6 months) caging experiment showed that 5 common species of sessile animals-3 sponges and 2 compound ascidians-were significantly reduced by seastar predation. Nevertheless, the amount eaten, considered with seastar densities and distributions, indicates that the seastars are unimportant in influencing the utilization of space by sessile fauna. From comparing caged and uncaged controls, we tentatively suggest that other predators are also unimportant, and that competition for substrate space and for access to the water column is (and, has been during the evolution of these sessile organisms) far more influential.     

The impact of behavioral and physiological maternal effects on offspring sex ratio in the common snapping turtle,<Emphasis Type="Italic"> Chelydra serpentina</Emphasis>

Theory suggests that maternal effects are especially important in organisms with environmentally-sensitive sex-determining mechanisms. However, there is no substantive body of empirical evidence to confirm this conjecture. We integrated field and laboratory studies to jointly evaluate the significance of behavioral (nest-site choice) and physiological (yolk hormone allocation) maternal effects on offspring sex ratio in the common snapping turtle (Chelydra serpentina), a species with temperature-dependent sex determination (TSD). Of the 16 microhabitat variables measured, only three (south, east, and total overstory vegetation cover) were significantly correlated with nest temperature: cooler nests were located under more vegetation cover. In turn, these microhabitat predictors of nest temperature, and nest temperature itself, may influence nest sex ratio: shadier, cooler nests were more likely to produce a higher proportion of male offspring than less shady, warmer nests. Analysis of eggs from these same nests incubated in a common garden design in the laboratory revealed that clutch sex ratio was unaffected by levels of yolk estradiol, yolk testosterone, or their interaction. Examination of both behavioral and physiological maternal effects revealed no concordant impact on offspring sex ratio. However, eggs from nests that produced male-biased sex ratios in the field yielded higher proportions of males under constant-temperature conditions in the laboratory. Our study confirms the importance of behavioral maternal effects in nature on offspring sex ratios in species with TSD, while also revealing the potential presence of a predisposition for sex-ratio production underlying TSD in this system.Communicated by S. Krackow     

Palaeoenvironmental controls on the distribution of Cretaceous herbivorous dinosaurs

Previous attempts to determine palaeoenvironmental preferences in dinosaurs have generally been qualitative assessments based upon data from restricted geographical areas. Here, we use a global database of Cretaceous herbivorous dinosaurs to identify significant associations between clades and broad palaeoenvironmental categories (‘terrestrial’, ‘coastal’, ‘marine’). Nodosaurid ankylosaurs and hadrosaurids show significant positive associations with marine sediments, while marginocephalians (Ceratopsia, Pachycephalosauria), saurischians (herbivorous theropods, Sauropoda) and ankylosaurid ankylosaurs are significantly positively associated with terrestrial sediments. These results provide quantitative support for the hypothesis that some clades (Nodosauridae, Hadrosauridae) were more abundant in coastal and/or fluvial environments, while others (e.g. Marginocephalia, Ankylosauridae) preferentially inhabited more distal environments. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. An erratum to this article can be found at