Reduction of sera requirements in amniotic fluid cell culture

Reduction in serum requirement for culture of primary human amniotic fluid cells can be achieved by the addition of 10 growth-promoting factors to the nutrient medium. This supplemented medium preserves cell types normally found in amniotic fluid cell cultures supplemented with 20–30 per cent fetal bovine serum. The volume of amniotic fluid required to initiate culture can be as little as 1 ml. Amniotic fluid samples contaminated with red blood cells with no visible clot also grow well in the low serum medium. Cell-free amniotic fluid combined with equal parts of supplemented medium is useful in initiating cell culture.     

The value of virtual conferencing for ecology and conservation

The objectives of conservation science and dissemination of its research create a paradox: Conservation is about preserving the environment, yet scientists spread this message at conferences with heavy carbon footprints. Ecology and conservation science depend on global knowledge exchange—getting the best science to the places it is most needed. However, conference attendance from developed countries typically outweighs that from developing countries that are biodiversity and conservation hotspots. If any branch of science should be trying to maximize participation while minimizing carbon emissions, it is conservation. Virtual conferencing is common in other disciplines, such as education and humanities, but it is surprisingly underused in ecology and conservation. Adopting virtual conferencing entails a number of challenges, including logistics and unified acceptance, which we argue can be overcome through planning and technology. We examined 4 conference models: a pure‐virtual model and 3 hybrid hub‐and‐node models, where hubs stream content to local nodes. These models collectively aim to mitigate the logistical and administrative challenges of global knowledge transfer. Embracing virtual conferencing addresses 2 essential prerequisites of modern conferences: lowering carbon emissions and increasing accessibility for remote, time‐ and resource‐poor researchers, particularly those from developing countries.     

Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

LEHR superfund stormwater runoff and putah creek mercury issues

Stormwater runoff from the University of California, Davis/U.S. Department of Energy Laboratory for Energy‐Related Health Research (UCD/US DOE LEHR) Superfund site located on the University of California campus in Davis, California, has been found to contain over 500 ng/L of total recoverable mercury, which is about ten times the California Toxics Rule criterion. This stormwater runoff is discharged to Putah Creek, which is Clean Water Act Section 303(d) listed as impaired for excessive mercury bioaccumulation in edible fish. A discussion is presented on the potential impact of the mercury in stormwater runoff from LEHR leading to excessive bioaccumulation of mercury in Putah Creek fish. The mercury in the stormwater runoff is derived from former flooding of the soils near the creek, which contains mercury derived from abandoned upstream mercury mines located in the Coast Range Vaca Hills to the west of LEHR. The implications of this situation for implementing a Total Maximum Daily Load (TMDL) to control mercury in stormwater runoff to Putah Creek are presented. © 2009 Wiley Periodicals, Inc.     

Application of biomarkers to assess the condition of European Marine Sites

A series of European Marine Sites has been designated as Special Areas of Conservation (SAC) in England. The aim of this study was to develop a practical methodology to assess the condition of SACs by applying a suite of biomarkers. Biomarkers were applied to the blue mussel Mytilus edulis and the shore crab Carcinus maenas from the Fal and Helford SAC (Cornwall). Individual biomarkers provided useful diagnostic information on the activity of certain classes of contaminants and an integrated Biomarker Response Index (BRI) was used to achieve a more holistic understanding of the condition of the SAC. The BRI indicated that the general health of both organisms was impacted in the upper part of the SAC (Fal Estuary) which correlated well with known chemical hotspots and sources of contamination. The BRI allows a pragmatic way to prioritise SAC sites that may require further investigative studies.     

Biological and economic optima in a tropical grazing ecosystem in Australia

Our aim in this study was to identify the economically optimal level of use of resources for a grazing system in tropical Australia and to compare it with the biological optimum. Grazing management trials were conducted in the Ord River irrigation area of north-western Australia. Biologists at the Commonwealth Scientific and Industrial Research Organization provided the biological data. The liveweight gains of Kimberley shorthorn steers were recorded, after a 1-year fattening period, for different grazing management strategies. Five different stocking rates were used on irrigated pangola grass (Digitaria decumbens) fertilized with nitrogen at five different rates. All other were applied at fixed, non-limiting levels.The results of the trials were modelled by a liveweight gain production function. This production function was used in an income simulation model to identify economically optimal (profit maximizing) stocking rates. This was done in two stages. First, a base analysis was undertaken on the basis o the economic conditions prevailing in September 1987. Second, sensitivity analysis was applied to the results of the base analysis. The parameters varied were the rate of interest, store cattle prices, finished cattle prices, transport costs and the length of the planning horizon.The biological optimum, when defined as maximum liveweight gain ha⁻¹, corresponds to a much higher intensity of use of resources than the economic optimum, and yields substantially lower profits or higher losses. These discrepancies increase when economic conditions worsen, or when the planning horizon of graziers is decreased from 50 to 10 years. Therefore, the adoption of the biological optimum cannot result in an econonomically stable grazing system in the medium to long term, when it is highly likely that economic parameters will fluctuate. The economic stability of the system is increased, but is still quite weak, if the economically optimal input combination is utilized. It was not possible to determine whether this economic optimum would result in long-term ecological stability because the relevant data were not collected during the trials.The relevance of management recommendations made to graziers and land administrators would be increased if these recommendations were based on an analysis of both the economic and the ecological stability of grazing systems over the medium to long term. We show in the last part of this study how this could be accomplished.     

Cover crop effects on nitrous oxide emission from a manure-treated Mollisol

Agriculture contributes 40–60% of the total annual N₂O emissions to the atmosphere. Development of management practices to reduce these emissions would have a significant impact on greenhouse gas levels. Non-leguminous cover crops are efficient scavengers of residual soil NO₃, thereby reducing leaching losses. However, the effect of a grass cover crop on N₂O emissions from soil receiving liquid swine manure has not been evaluated. This study investigated: (i) the temporal patterns of N₂O emissions following addition of swine manure slurry in a laboratory setting under fluctuating soil moisture regimes; (ii) assessed the potential of a rye (Secale cereale L.) cover crop to decrease N₂O emissions under these conditions; and (iii) quantified field N₂O emissions in response to either spring applied urea ammonium nitrate (UAN) or different rates of fall-applied liquid swine manure, in the presence or absence of a rye/oat winter cover crop. Laboratory experiments investigating cover crop effects N₂O emissions were performed in a controlled environment chamber programmed for a 14 h light period, 18 °C day temperature, and 15 °C night temperature. Treatments with or without a living rye cover crop were treated with either: (i) no manure; (ii) a phosphorus-based manure application rate (low manure): or (iii) a nitrogen-based manure application rate (high manure). We observed a significant reduction in N₂O emissions in the presence of the rye cover crop. Field experiments were performed on a fine-loamy soil in Central Iowa from October 12, 2005 to October 2, 2006. We observed no significant effect of the cover crop on cumulative N₂O emissions in the field. The primary factor influencing N₂O emission was N application rate, regardless of form or timing. The response of N₂O emission to N additions was non-linear, with progressively more N₂O emitted with increasing N application. These results indicate that while cover crops have the potential to reduce N₂O emissions, N application rate may be the overriding factor.     

Principles and issues in radiological ecological risk assessment

This paper provides a bridge between the fields of ecological risk assessment (ERA) and radioecology by presenting key biota dose assessment issues identified in the US Department of Energy's Graded Approach for Evaluating Radiation Doses to Aquatic and Terrestrial Biota in a manner consistent with the US Environmental Protection Agency's framework for ERA. Current radiological ERA methods and data are intended for use in protecting natural populations of biota, rather than individual members of a population. Potentially susceptible receptors include vertebrates and terrestrial plants. One must ensure that all media, radionuclides (including short-lived radioactive decay products), types of radiations (i.e., alpha particles, electrons, and photons), and pathways (i.e., internal and external contamination) are combined in each exposure scenario. The relative biological effectiveness of alpha particles with respect to deterministic effects must also be considered. Expected safe levels of exposure are available for the protection of natural populations of aquatic biota (10 mGy d(-1)) and terrestrial plants (10 mGy d(-1)) and animals (1 mGy d(-1)) and are appropriate for use in all radiological ERA tiers, provided that appropriate exposure assumptions are used. Caution must be exercised (and a thorough justification provided) if more restrictive limits are selected, to ensure that the supporting data are of high quality, reproducible, and clearly relevant to the protection of natural populations.