Determination of toxic cyclic heptapeptides by liquid chromatography with detection using ultra-violet, protein phosphatase assay and tandem mass spectrometry

Microcystins, toxic cyclic heptapeptides and nodularin-R, a toxic cyclic pentapeptide, were determined using liquid chromatography (LC) with detection using photo-diode array ultra-violet (PDA-UV) and protein phosphatase (PP) assay. Positive fractions were analysed for toxins using collision-induced dissociation (CID) and tandem MS/MS experiments which were carried out simultaneously using electrospray ion-trap instrumentation. Reversed-phase liquid chromatography (LC) using an acetonitrile/water gradient was used for the LC-MS(2) determination of six microcystins standards and nodularin. The molecular related ion species, [M+H](+)([M+2H](2+) in the case of MC-RR), were used as the precursor ions for MS(2) experiments. Optimum calibration and reproducibility data were obtained for MC-LR using LC-MS(2); 0.1-5.0 microg/ml, r2 = 0.992 (n = 3); % RSD < or =7.3 at 0.25 microg MC-LR/ml (n = 3). The detection limit (S/N = 3) was better than 0.1 ng. Water samples for microcystin analysis were first screened using protein phosphatase (PP) assays and positives were concentrated using C-18 solid-phase extraction. The developed method was applied to examine a lake in Ireland contaminated by Microcystis sp. and MC-LR and MC-LA were identified.     

Phytoremediation of petroleum hydrocarbons in tropical coastal soils II. microbial response to plant roots and contaminant

GOAL, SCOPE AND BACKGROUND: The goal of this study was to understand the interaction between plants and microorganisms during petroleum-hydrocarbon bioremediation in Pacific Islands coastal soils. Total bacteria and hydrocarbon-degrading microorganisms population dyanamics were examined in the rhizospheres of tropical trees and shrubs, which were evaluated for their phytoremediation potential in a greenhouse experiment. The respective and combined effects of plant roots and diesel contaminant on the microbial populations were determined in relation to diesel fuel depletion. An increase in the grading populations size of the hydrocarbon-degrading populations of microbes, elicited by rhizodeposition, is generally regarded as conducive to an enhanced degradation of petroleum hydrocarbon pollutants in vegetated soil. METHODS: The soil was a coastal sandy loam (pH 7.8) which was artificially contaminated with 10 g of No. 2 diesel fuel/kg soil or left uncontaminated. The pots were irrigated with fertilizer and 1% NaCl. The enumerations were carried out in the contaminated and uncontaminated rhizospheres of three trees, kiawe (Prosopis pallida), milo (Thespesia populnea), and kou (Cordia subcordata) and three shrubs, beach naupaka (Scaevola sericea), false sandalwood (Myoporum sandwicense), and oleander (Nerium oleander). Unplanted control soils were included in the experiment. Total bacteria and phenanthrene-degrading bacteria were enumerated on plates. Diesel- and pristane-degrading microorganisms were enumerated by the most-probable-number technique in tissue-culture plates. RESULTS AND DISCUSSION: All four types of microorganisms responded to the rhizosphere of the 6 plants in uncontaminated soil and to the diesel contaminant in unplanted soil. In contaminated rhizospheres, no effect of the plant on the hydrocarbon-degrader numbers was visible. Total bacteria responded more to the plant roots than to the contaminant. The phenanthrene-degrading bacteria and pristane-degrading microorganisms were more influenced by the contaminant than by the plants. The diesel-degrading microorganisms were equally stimulated by the plants and the contaminant. The numbers of hydrocarbon degraders were similar in the contaminated rhizospheres of the three effective plants (kiawe, kou, and milo) and in those of the three ineffective shrubs. CONCLUSION: The results suggest the quality of the rhizodeposition is plant-dependent and governs the type of diesel-degrader populations that will be enhanced by a given plant. RECOMMENDATIONS AND OUTLOOK: In the proposed phytoremediation-benefit model plant roots maintain high levels of hydrocaron degraders in uncontaminated soil. When the root enters a contaminated zone of soil, those hydrocarbon degraders that prefer the contaminant would switch to the contaminant as a carbon source, effectively removing the hydrocarbons. If the root exudates and the contaminant are equally attractive to the hydrocarbon degraders, the contaminant degradaton would be less effective.     

Partitioning of dioxins and dibenzofurans: Whole blood, blood plasma and adipose tissue

It has become relatively common to estimate human dioxin body burden and to document dioxin exposures by measuring dioxin and dibenzofuran congeners in adipose tissue, whole blood or blood plasma, and reporting these values on a lipid basis. It has not been determined whether these three types of specimens contain identical dioxin and dibenzofuran levels. This paper compares paired plasma and adipose tissue and paired whole blood and adipose tissue in analyses from two groups of patients. The first group consists of twenty U.S. veterans with paired plasma and adipose specimens. The second group consists of four German adults with whole blood compared to adipose tissue. Forty-eight analyses were performed. The results suggest that for some higher chlorinated compounds, such as OCDD, plasma lipid values may be higher than adipose lipid values, but whole blood lipid values for the higher as well as lower chlorinated PCDD/Fs may be relatively similar. On the other hand, the values for the lower chlorinated PCDD/Fs, such as TCDD, are similar in blood plasma, adipose tissue and whole blood. Total PCDD/F dioxin “toxic equivalents” are similar within each of the two series reported here, using the current “International Dioxin Toxic Equivalent” system.     

Improved light extinction reconstruction in interagency monitoring of protected visual environments

The U.S. Environmental Protection Agency (EPA) published the Regional Haze Rule (RHR) in 1999. The RHR default goal is to reduce haze linearly from the baseline period of 2000 through 2004 to natural background in 2064. EPA-recommended method for estimating baseline and natural haze uses the Interagency Monitoring of Protected Visual Environments (IMPROVE) light extinction formula. The IMPROVE formula predicts light extinction from measured aerosol chemical concentrations and estimates of the relative humidity multiplier. On average, the IMPROVE formula overpredicts 6156 nephelometer days (24-hr average measured particle light scattering, bsp) of data by 25%. A new IMPROVED method that reconstructs light extinction using a concentration power law model overpredicts these nephelometer days of data by just 2%. Ignoring the 20% lowest light scattering days, this new IMPROVED formula has a 3% underprediction bias over the 4925 highest nephelometer days with light scattering > or =8 inverse megameters. For comparison, the IMPROVE formula has a 12% overprediction bias for the same days. The IMPROVE formula overprediction averages 77%, 27%, 17%, 9%, and -5% broken down by quintile from lowest to highest nephelometer measured light scattering days. The new IMPROVED formula average overprediction is 21%, -5%, -5%, -2%, and 0%. So, agreement between measured and predicted light scattering improves by modifying the current IMPROVE light extinction formula.     

Ozone dynamics and deposition processes at a deforested site in the Amazon basin

The goal of this study is to investigate the impact of deforestation on ozone dynamics and deposition in the Brazilian Amazon basin. This goal is accomplished through i) analyses of ozone levels and deposition rates at a deforested site during the rainy season; and ii) comparisons of these data with similar information derived at a forest. At the pasture site maximum ozone mixing ratios reach 20 parts per billion on a volume basis (ppbv) but about 6 ppbv prevail over the forest. Maximum ozone deposition velocities for pastures can reach 0.7 cm s-1, which is about threefold lower than values derived for forests. Combining ozone abundance and deposition velocities, pasture maximum ozone fluxes reach approximately 0.2 microgram (ozone) m-2 s-1. This flux represents approximately 70% of the deposition rates measured over the forest. Hence, this study suggests that conversion of rainforests to pastures could lead to a net reduction (30%) in the ozone sink in the Amazon.     

Toxicological response and its reversibility in rats fed Lake Ontario or Pacific coho salmon for 13 weeks

Lake Ontario coho salmon were known to contain a mixture of chemical contaminants. A previous study demonstrated that rats fed the Lake Ontario fish-supplemented diet for 28 days exhibited mild biochemical and histological changes. The purpose of the present study was to determine the effects due to a longer term of exposure and the reversibility of these effects. Growth rate and food consumption were not affected by feeding the animals with Lake Ontario or Pacific fish-supplemented diets for 13 weeks. No deaths were observed. Decreased spleen weights were observed in groups of males fed 1.45%, 5.8% Lake Ontario and 2.9% Pacific diet. After a 13 week recovery the spleen weights returned to normal. Decreased serum potassium was observed in male rats fed 2.9% Lake Ontario diet, and all levels of Pacific diet for 13 weeks, and was not evident following maintenance on normal diet. Serum glucose was not affected by the 13-week period of treatment, however; a reduction in this parameter occurred in male rats fed the two highest doses of Lake Ontario diet and all doses of Pacific diet following the 13-week recovery period. Minor hematological changes occurred only in the male rats fed either Lake Ontario or Pacific diet following a 13 week recovery period and included reduced marrow myeloid cells and myeloid/erythroid ratio. Hepatic microsomal ethoxyresorufin deethylase activity was significantly increased in rats ingesting Lake Ontario diet. Mild histological changes occurred in the liver and thyroid of the treated males, and in the liver and kidney of the treated females. These changes were attributed to the chemical residues and/or the fish diet. Data presented here indicated that the Lake Ontario fish-supplemented diet can cause mild biochemical, hematological and histological changes but most of these were reversible when exposure was terminated.     

Concentrations of dioxin-like PCB congeners in unweathered Aroclors by HRGC/HRMS using EPA Method 1668A

We have determined the congener compositions of nine commercial Aroclor products of polychlorinated biphenyls (PCBs) to the sub-part-per-million level using high-resolution gas chromatography combined with high-resolution mass spectrometry according to US Environmental Protection Agency (EPA) Method 1668A. These Aroclor composition data should allow improved characterization and risk assessment of PCB contamination at hazardous waste sites, particularly for dioxin-like PCB congeners. By combining the data on the concentrations of each dioxin-like congener with its World Health Organization toxicity equivalency factor, we have established dioxin toxic equivalent concentrations for each pure Aroclor product.     

Sexual selection on alleles that determine body size in the swordtail Xiphophorus nigrensis

Summary In a natural population, we document changes in the frequencies of alleles influencing body size and size-correlated alternative male reproductive tactics, and we examine the possible role of sexual selection in producing these changes. Male swordtails (Xiphophorus nigrensis, Rio Choy) exhibit three body size classes (small, intermediate, and large) that primarily derive from allelic variation (s, I, L) at the Y-linked pituitary (P) locus. Some XX individuals are male. They are small and can be fathered either by XX or XY males. We compared the frequencies of Y-linked P alleles across two generations in a natural population. There was a significant decrease in the Y-s genotype relative to Y-I and Y-L genotypes. Laboratory experiments suggest that the disadvantage of Y-s resulted, at least in part, from female preference for larger courting males relative to small chasing males. All larger courting males are of the Y-I or Y-L genotypes. Although the frequency of Y-linked P alleles changed across generations, there was no change in the distribution of male body size classes. The stability of the phenotypic distribution across generations, despite changes in P allele frequencies, was maintained by the production of small XX sons by Y-I and Y-L males. Offprint requests to: M.J. Ryan     

Discrimination between nestmate and non-nestmate kin by social wasps (Polistes fuscatus,Hymenoptera: Vespidae)

Summary The threespine stickleback,Gasterosteus aculeatus is a voracious cannibal of both its own eggs and those of conspecific neighbours. Females, but not males, can distinguish their progeny from those of other fish and attacked alien eggs more frequently. In experiments to examine nest raiding, females that initiated raids on nests resulting in cannibalism were the first females to spawn in the reconstructed nest. These results support the hypothesis that cannibalism by females may be adaptive in situations where intense female competition for male guardians occurs.     

Robust benchmark dose determination based on profile score methods

We investigate several methods commonly used to obtain a benchmark dose and show that those based on full likelihood or profile likelihood methods might have severe shortcomings. We propose two new profile likelihood-based approaches which overcome these problems. Another contribution is the extension of the benchmark dose determination to non full likelihood models, such as quasi-likelihood, generalized estimating equations, which are widely used in settings such as developmental toxicity where clustered data are encountered. This widening of the scope of application is possible by the use of (robust) score statistics. Benchmark dose methods are applied to a data set from a developmental toxicity study.