Biacore biosensor immunoassay for 4-nonylphenols: assay optimization and applicability for shellfish analysis

A rapid Biacore biosensor immunoassay of 4-nonylphenols was developed. Two types of antibodies were used in the study: polyclonal antibodies with high cross-reactivity towards technical 4-nonylphenol and a monoclonal antibody very specific to 4-n-nonylphenol. 9-(p-Hydroxyphenyl)nonanoic acid was immobilized onto surface of a sensor chip. The best assay sensitivity was achieved using a flow rate of 50 microl min(-1) and injection time of 2 min. For the assay incorporating monoclonal antibodies a limit of detection 2 ng ml(-1) for 4-n-nonylphenol was achieved. With polyclonal antibodies one order lower sensitivity was observed for 4-nonylphenols. High background level of calibration curve for technical 4-nonylphenol was decreased by using IgG fraction of polyclonal antibodies in combination with lower amount of immobilised 9-(p-hydroxyphenyl)nonanoic acid. Sensitivity of the assay was improved by using a chip with a new derivative on a surface-N-aminobutyl [2-(4-hydroxyphenyl)ethylamine] (limit of detection--5 ng ml(-1)). Applicability of the developed assays to ecological monitoring was checked in experiments using shellfish samples. 4-n-Nonylphenol from spiked samples was extracted into hexane followed by clean-up on NH2 SPE columns. Calibration curves generated for cockles, mussels and oyster samples were identical (limit of detection about 10 ng g(-1)) whereas for scallop samples a slight decrease (about 5-10%) of absolute response was observed. In the assay using the monoclonal antibody specific to 4-n-nonylphenol 31 shellfish samples were found to be negative. Results obtained with polyclonal antibodies indicated that two scallop samples contained a quantity of 4-nonylphenols. The developed biosensor assay could be applied for shellfish analysis as a preliminary screening method.     

Host selection,location, and association behaviors of anemonefishes in field settlement experiments

Nine anemonefish species were reared in the laboratory, and individuals were released in the field (Lizard Island, Australia and Madang, Papua New Guinea) at different distances and orientations away from natural (anemone species the fishes are found with in nature) and unnatural species of host anemones. Experiments were conducted to examine factors that could affect the settlement behaviors of the fishes: current velocity, distance and orientation of the fishes to the anemones, chemical vs visual cues, and presence of conspecific or heterospecific fishes. The fishes were usually attracted toward natural host species of anemones but not towards unnatural host anemone species nor to pieces of dead coral. Host selection during settlement provided the best explanation for the host specificity patterns displayed by anemonefishes in nature. The fishes used chemical cues released from the anemones to identify and locate the appropriate host species and could effectively locate the anemones from a maximum distance of 8 m downstream. Fishes released upstream or to the side of anemones (where anemone chemicals were assumed to be reduced or absent) were much less successful in locating anemones. The ability of the fishes to locate natural host anemones was strongly reduced when there was no water current. The presence of resident anemonefishes on host species of anemones did not influence the attraction behavior of anemonefishes released downstream from the anemones. Once the released fishes got close to or entered the anemones, the resident fishes would generally bite and chase them until the recruits left the anemones. Most fishes were not stung upon initial contact with the anemones.     

Mercury-selenium-bromine imbalance in premature parturient California sea lions

High premature birth rates have been observed in the rookeries of the California sea lion Zalophus californianus since 1968. The reasons for the premature pupping are complex and, hence, not well understood, although leptospirosis infection and elevated PCB and DDT residues have been implicated. We were interested in determining what role trace and major elements played in these events. Livers and kidneys from 10 normal parturient and 10 premature parturient mothers and their pups were analyzed for Hg, Se, Br, Cd, Ag, Cu, Fe, Zn, Mn, K, Na, Ca, and Mg in order to detect differences that might exist between the two groups. A further objective was to establish how these elements varied in relation to each other in the normal and abnormal sea lions. Our results revealed that Hg, Se, Cd, and Br levels were significantly higher in the livers of the normal mothers and that these elements were all in balance (highly correlated) with each other. This was especially true for Hg, Se, and Br. In mothers with high concentrations of these elements (e.g. Hg greater than 800 g/g dry weight), atomic ratios of approximately 1Hg:1Se:1Br were observed. Atomic Se:Hg ratios were also near unity in the abnormal mothers; however, Br concentrations were always severely depressed in these individuals. Normal full-term pups had higher hepatic levels of Hg and Se, and near-perfect 1:1 Se:Hg atomic ratios were almost always observed. In contrast, the livers of the premature pups appeared to be deficient in Hg, and, consequently, elevated Se:Hg ratios were always found. In almost all cases, the premature pups had increased concentrations of Na, Ca, and Br. Levels of these elements were correlated with their Se:Hg ratios. Amounts of Mn and Cu were reduced in the premature pups and negatively correlated with Se:Hg ratios. The results suggest that balance between elements is of more importance than absolute concentration when the possible effects of toxic elements are considered. It also appears that bromine may be important in the detoxification process involving Se and Hg and perhaps Cd as well; i.e., every mother that had Br in balance with Hg, Cd, and Se had a normal pup, while every mother that lacked sufficient Br had a premature pup. The question of whether Hg detoxifies Se is also raised. All the normal pups had Se:Hg atomic ratios of less than 2.2, while all the premature pups had reduced Hg amounts and Se:Hg ratios above 3.4.     

Fathead Minnow and Bluegill Sunfish Life‐Stage Responses to 17β‐Estradiol Exposure in Outdoor Mesocosms

Developmental and reproductive effects of 17β‐estradiol (E2) exposure on two generations of fathead minnows and one generation of bluegill sunfish were assessed. Fish were exposed to E2 for six continuous weeks in outdoor mesocosms simulating natural lake environments. First generation fish were exposed while sexually mature. Second generation fathead minnows were exposed either during early development, sexual maturity, or both stages. Multiple endpoints were measured to assess effects of E2 exposure on fecundity and fish health and development. Plasma vitellogenin concentrations were highly variable in all fish. Differences in egg production timing for both species indicate differences in fecundity between females exposed to E2 and controls. First generation fathead minnows exposed to E2 had lower body condition factors and reduced secondary sexual characteristic expression by males. Only a difference in relative liver weight was observed in second generation fathead minnows. First generation bluegill males exposed to E2 had significantly smaller testes compared to controls. Although fish response was highly variable, results indicate that exposure to E2 at environmentally relevant concentrations affect fathead minnow and bluegill sunfish health and development, which may have implications for the health and sustainability of fish populations. Furthermore, exposure timing and environmental factors affect fish response to E2 exposure.     

Extrinsic regime shifts drive abrupt changes in regeneration dynamics at upper treeline in the Rocky Mountains, U.S.A

Given the widespread and often dramatic influence of climate change on terrestrial ecosystems, it is increasingly common for abrupt threshold changes to occur, yet explicitly testing for climate and ecological regime shifts is lacking in climatically sensitive upper treeline ecotones. In this study, quantitative evidence based on empirical data is provided to support the key role of extrinsic, climate-induced thresholds in governing the spatial and temporal patterns of tree establishment in these high-elevation environments. Dendroecological techniques were used to reconstruct a 420-year history of regeneration dynamics within upper treeline ecotones along a latitudinal gradient (approximately 44-35 degrees N) in the Rocky Mountains. Correlation analysis was used to assess the possible influence of minimum and maximum temperature indices and cool-season (November-April) precipitation on regional age-structure data. Regime-shift analysis was used to detect thresholds in tree establishment during the entire period of record (1580-2000), temperature variables significantly Correlated with establishment during the 20th century, and cool-season precipitation. Tree establishment was significantly correlated with minimum temperature during the spring (March-May) and cool season. Regime-shift analysis identified an abrupt increase in regional tree establishment in 1950 (1950-1954 age class). Coincident with this period was a shift toward reduced cool-season precipitation. The alignment of these climate conditions apparently triggered an abrupt increase in establishment that was unprecedented during the period of record. Two main findings emerge from this research that underscore the critical role of climate in governing regeneration dynamics within upper treeline ecotones. (1) Regional climate variability is capable of exceeding bioclimatic thresholds, thereby initiating synchronous and abrupt changes in the spatial and temporal patterns of tree establishment at broad regional scales. (2) The importance of climate parameters exceeding critical threshold values and triggering a regime shift in tree establishment appears to be contingent on the alignment of favorable temperature and moisture regimes. This research suggests that threshold changes in the climate system can fundamentally alter regeneration dynamics within upper treeline ecotones and, through the use of regime-shift analysis, reveals important climate-vegetation linkages.     

Improving species status assessments under the U.S. Endangered Species Act and implications for multispecies conservation challenges worldwide

Despite its successes, the U.S. Endangered Species Act (ESA) has proven challenging to implement due to funding limitations, workload backlog, and other problems. As threats to species survival intensify and as more species come under threat, the need for the ESA and similar conservation laws and policies in other countries to function efficiently has grown. Attempts by the U.S. Fish and Wildlife Service (USFWS) to streamline ESA decisions include multispecies recovery plans and habitat conservation plans. We address species status assessment (SSA), a USFWS process to inform ESA decisions from listing to recovery, within the context of multispecies and ecosystem planning. Although existing SSAs have a single-species focus, ecosystem-based research can efficiently inform multiple SSAs within a region and provide a foundation for transition to multispecies SSAs in the future. We considered at-risk grassland species and ecosystems within the southeastern United States, where a disproportionate number of rare and endemic species are associated with grasslands. To initiate our ecosystem-based approach, we used a combined literature-based and structured World Café workshop format to identify science needs for SSAs. Discussions concentrated on 5 categories of threats to grassland species and ecosystems, consistent with recommendations to make shared threats a focus of planning under the ESA: (1) habitat loss, fragmentation, and disruption of functional connectivity; (2) climate change; (3) altered disturbance regimes; (4) invasive species; and (5) localized impacts. For each threat, workshop participants identified science and information needs, including database availability, research priorities, and modeling and mapping needs. Grouping species by habitat and shared threats can make the SSA process and other planning processes for conservation of at-risk species worldwide more efficient and useful. We found a combination of literature review and structured discussion effective for identifying the scientific information and analysis needed to support the development of multiple SSAs. Article impact statement: Species status assessments can be improved by an ecosystem-based approach that groups imperiled species by shared habitats and threats.     

Pathways for the oxidation of sarin in urban atmospheres

The nerve agent sarin has recently been deployed by terrorists in a major city. The molecule is volatile and made its way to many victims by passing as vapor through a highly reactive medium. Here we estimate rates and pathways for the removal of gas phase sarin from a generalized urban atmosphere. Only information from the open scientific literature is used. By structure reactivity comparisons with the organophosphorus pesticides, hydroxyl radical hydrogen abstraction may occur in as little as one hour. Decomposition of side chains after hydroxyl attack leads to organic oxygenates which preserve the phosphonofluoridate and so toxicity. The aqueous aerosol surface is contacted in minutes and offers access to a range of dissolved nucleophiles. Substitution displaces the fluoride leaving group, giving safe phosphoric acid analogs. Because of uncertainties in the electron distribution and in aqueous decay mechanisms, the time constants must be viewed as lower limits.     

Determinants of homeowners' attitudes to the installation of high-voltage overhead transmission lines

This article deals comprehensively with factors which influence attitudes to the placement of high-voltage overhead transmission lines (HVOTLs). It employs an analytical schema which links the initial stimulus of the power lines' proposal in an area to Background and Socio-economic independent variables and a set of mediating variables. From a survey of 600 residents in three different domiciliary settings in Queensland, Australia, results indicate an absence of positive attitudes and, at best, neutral ones to HVOTL placement. By multinomial logistic regression, the study probes the grounds behind negativity and extreme negativity, uncovering direct links between these attitudes and certain of the variables included in the analytical schema.