David K. Ryan

Toxicity of the 13 priority pollutant metals to Vibrio fisheri in the Microtox® chronic toxicity test

The Microtox Acute Toxicity Test has been successfully used to measure the toxicity of metals and other pollutants at high concentrations (ppm) in selected environmental samples. However, metals and other toxicants are often found in much lower concentrations (ppb) in many municipal wastewaters and receiving waters. In order to assess the toxicity of these pollutants in these samples, a more sensitive toxicity assay is needed. The Microtox chronic toxicity test has been developed to measure the sublethal effect of toxicants over multiple generations of the test species, Vibrio fisheri. In this study, the toxicity of the 13 priority pollutant metals [i.e. As, Se, Cd, Cr (III and VI), Cu, Pb, Sb, Ag, Tl, Zn, Be, Hg and Ni] to V. fisheri was evaluated using the Microtox chronic toxicity test. In this test, the inhibitory concentration (IC), lowest observable effect concentration (LOEC), and no observable effect concentration (NOEC) were obtained after 22-h of incubation at 27+/-1 degrees C, by comparing the light output of the control to that of the test sample. Among the 13 priority pollutant metals, beryllium (Be) was found to be the most toxic in the test (LOEC=0.742-1.49 microg/l) while thallium (Tl) was the least toxic (LOEC=3840-15300 microg/l). The LOECs for copper (as Cu) and lead (Pb) in reagent (ASTM Type I) water were 6.78-13.6 microg/l and 626-1251 microg/l, respectively. The toxicity of copper sulfate (as Cu) in reagent water was shown and significantly reduced with the addition of natural organic matter (fulvic acid) or EDTA to the sample. The LOEC values for the 13 priority pollutant metals in this test were comparable to or lower than those reported for commonly used aquatic toxicity tests, such as the Ceriodaphnia dubia assay.

Copper(II) complexing capacities of natural waters by fluorescence quenching.

The natural fluorescence of humic substances in both fresh waters and marine samples is quenched upon complexation to Cu2+ ion. Binding curves obtained from titration data are mathematically modeled and complexing capacities ( CL) determined by computer curve fitting. The results indicate that the fluorescence technique directly measures organic matter complexation only, and CL values are unaffected by hydrolysis. Filtered water samples are analyzed at their natural pH under N2 but are otherwise unaltered. Rayleigh scattering, measured along with fluorescence, signals precipitate formation and was used to indicate a suitable stopping point for titrations. Scattering and fluorescence trends were similar for all samples except an estuarine sample. A multiple correlation study of several commonly measured water characteristics and the titration parameters showed a statistically significant trend between UV absorbance and CL values.

Atmospheric deposition of toxics onto Massachusetts Bay-I. Metals

Wet and dry atmospheric deposition of toxic metals was measured at biweekly intervals for one year, from i5 September i992 to i6 September i993 at two sites on Massachusetts Bay, Nahant, near Boston and Truro, near the tip of Cape Cod. Wet and dry deposition was measured using a conventional wet/dry collector, except that the dry bucket contained a layer of water in order to simulate the uptake of dry deposition onto a water surface. In addition, at Nahant, a dichotomous particle collector was used to measure metal concentrations on particles. Analytical methods were INAA and ICP-MS. Generally, dry deposition of metals was greater at Nahant than at Truro, and wet deposition was greater or equal at Truro than at Nahant. Averaging results from the two sites, the following deposition rates (wet + dry) were obtained for the Bay in μg m−2 yr−1: Al i02000, As i32, Cd 405, Co 58, Cr 2700, Cu 3500, Fe 140,000, Mn 4420, Ni 7200, Pb 2700, Sb i60, Se 264, Zn 7800. Preliminary results are also given for mercury wet deposition, which was measured for 6 weekly periods in the fall of 1993.

Atmospheric deposition of toxics onto Massachusetts Bay-II. Polycyclic aromatic hydrocarbons

Wet and dry atmospheric deposition of polycyclic aromatic hydrocarbons (PAHs) was measured at biweekly intervals from 15 September 1992 to 16 September 1993 at two sites on Massachusetts Bay, Nahant, near Boston and Truro, near the tip of Cape Cod. Wet and dry deposition was measured using a conventional wet/dry collector, except that the dry bucket contained a layer of water in order to simulate the uptake of dry deposition onto a water surface. The PAHs were extracted from the aqueous solution/suspension by methylene chloride, preconcentrated by low-temperature evaporation, and analyzed by GC-MS. Dry and wet depositions of PAHs were significantly greater at Nahant than at Truro, due to the proximity of emission sources in the metropolitan Boston area. Highest deposition of PAHs was observed in the winter season. At Nahant, the following deposition rates (wet + dry) were observed in units of 104 ng m−2 yr−1: fluoroanthene, 13; phenanthrene, 11; pyrene, 9.8; chrysene, 5.3; and the potentially carcinogenic benzo(a) pyrene, 3. At Truro, the deposition rates were several fold lower. Polychlorinated biphenyls (PCBs) were not found above the detection limit of the analytical procedure.

Gut contents: A significant contaminant of Mytilus edulis whole body metal concentrations

Ingested matter can have a significant effect on whole body metal concentration measurements in Mytilus edulis. Depuration of mussels in clean seawater for 36 h prior to dissection eliminates most of these contaminating gut contents. Depuration followed by metal analyses is the most direct method of determining mussel tissue metal bioburdens. After being transplanted into a plume of primary treated sewage effluent in Salem Harbor, Massachusetts for 32 days, Al, Cr, and Fe concentrations in depurated mussels were significantly lower than those determined for either non-depurated mussels or for depurated mussels to which fecal concentrations of Al, Cr, and Fe were added back in. Although mathematical methods developed by both Ouellette (1978) and Boehm et al. (1988) could be applied to non-depurated mussels in order to correct for errors associated with gut metal contamination, these indirect methods were not as reliable as depuration prior to analysis.

Lamin B Receptor Regulates the Growth and Maturation of Myeloid Progenitors via its Sterol Reductase Domain: Implications for Cholesterol Biosynthesis in Regulating Myelopoiesis

Lamin B receptor (LBR) is a bifunctional nuclear membrane protein with N-terminal lamin B and chromatin-binding domains plus a C-terminal sterol Δ14 reductase domain. LBR expression increases during neutrophil differentiation, and deficient expression disrupts neutrophil nuclear lobulation characteristic of Pelger-Huët anomaly. Thus, LBR plays a critical role in regulating myeloid differentiation, but how the two functional domains of LBR support this role is currently unclear. We previously identified abnormal proliferation and deficient functional maturation of promyelocytes (erythroid, myeloid, and lymphoid [EML]-derived promyelocytes) derived from EML-ic/ic cells, a myeloid model of ichthyosis (ic) bone marrow that lacks Lbr expression. In this study, we provide new evidence that cholesterol biosynthesis is important to myeloid cell growth and is supported by the sterol reductase domain of Lbr. Cholesterol biosynthesis inhibitors caused growth inhibition of EML cells that increased in EML-derived promyelocytes, whereas cells lacking Lbr exhibited complete growth arrest at both stages. Lipid production increased during wild-type neutrophil maturation, but ic/ic cells exhibited deficient levels of lipid and cholesterol production. Ectopic expression of a full-length Lbr in EML-ic/ic cells rescued both nuclear lobulation and growth arrest in cholesterol starvation conditions. Lipid production also was rescued, and a deficient respiratory burst was corrected. Expression of just the C-terminal sterol reductase domain of Lbr in ic/ic cells also improved each of these phenotypes. Our data support the conclusion that the sterol Δ14 reductase domain of LBR plays a critical role in cholesterol biosynthesis and that this process is essential to both myeloid cell growth and functional maturation.

Annual cycle of heavy metals in a tropical lake‐lake Chapala, Mexico

Abstract Dissolved concentrations of seven metals (As, Pb, Cd, Zn, Ni, Cu, Cr) were determined monthly at six sampling stations in Lake Chapala, Mexico, between November 1990 and December, 1991. Lake Chapala is the largest lake in Mexico, and serves as a source of drinking water and an economically important fishery and center for tourism. Historically, Lake Chapala has received poorly characterized domestic, industrial, and agricultural wastes. As, Cd, Pb, Cr, and Zn displayed peaks in concentration during the period before the rainy season due, presumably, to evaporation. Comparison of inflow and outflow concentrations also indicated a direct source of As to the lake. During this period, the concentrations of both Cd and Pb were above the freshwater chronic criteria values recommended by the United States Environmental Protection Agency for protection of aquatic ecosystem health. The metals As, Cr, Zn, Ni, and Cu also showed concentration peaks following the rainy season in August and September, perhaps...

A rapid fluorescence quenching method for the determination of equilibrium parameters for copper(II) complexation by humic materials

Abstract Fluorescence quenching measurements for determining stability constants ( K ) and complexation capacities ( C L ) of copper(II) complexation by humic materials have primarily been conducted by titrating a sample with metal ion. These titrations are long and tedious, requiring many data points, pH adjustment between additions, and long equilibration times. Once collected, the data must then be manipulated using regression analyses to obtain the equilibrium parameters. This paper describes the development of a new, rapid method of determining equilibrium parameters from fluorescence quenching. The use of individual pH-buffered aliquots of sample for each point along the titration curve yields data that can be analyzed with a simplified linear data treatment to obtain values for C L and K for the sample. This rapid fluorescence quenching method was evaluated using a model ligand (salicylic acid), isolated soil and stream fulvic acids, and a natural water sample.

Effects of emulsion viscosity during surfactant‐enhanced soil flushing in porous media

Surfactants can potentially improve the efficiency of pump‐and‐treat technology for remediation of aquifers contaminated by nonaqueous phase liquids (NAPLs). However, the formation of emulsions during the removal process can Increase the viscosity in the system. This can result in pore clogging and reduction of flow, which inhibits the contaminant removal process. Formation of viscous emulsions has been identified in previous research as one of the probable causes for in situ field test failures using surfactant‐enhanced soil‐flushing technology. However, the effects of in situ emulsification and viscosity increases have not been quantified previously. The purpose of this article is to investigate effects of in situ emulsification on the remediation process. Laboratory column studies examined the mobilization of m‐xylene from porous media using a 1% alcohol ethoxylate surfactant solution (Witconol® SN90). Effects of in situ emulsification were determined. Glass columns (1.1 cm i.d. × 30 cm) were packed wi...

Multi-Wavelength Fluorescence-Quenching Model for Determination of Cu2+ Conditional Stability Constants and Ligand Concentrations of Fulvic Acid:

In this work, a multi-wavelength model (MWM) is developed. It uses fluorescence bands in the fulvic acid (FA) spectrum that quench upon binding of inorganic Cu2+ to FA. Quenching data at pH values of 5, 6, and 7 are placed in sets, containing fluorescence measures at select wavelengths versus added copper (CM). Intensity data of wavelength set 1 are obtained from 25 nm constant offset synchronous fluorescence spectra (SyF), in which are observed distinct peaks (λex = 415 nm, λem = 440 nm; and λex = 471 nm, λem = 496 nm). Wavelength set 2 intensity data are obtained from the FA fluorescence excitation and emission maxima (λex = 335 nm, λem = 450 nm; and λex = 471 nm, λem = 496 nm). Application of MWM shows that the multi-wavelength data sets characterize ligands of different binding strength (log Kx) and concentration (CLx). Corresponding to pH values of 5, 6, and 7, mean and standard deviation values for wavelength set 1 are log K415/440 = 4.66 (0.12), 5.03 (0.12), and 5.05 (0.08), log K471/496 = 4.93 (0.06), 5.27 (0.11), and 5.39 (0.09), C415/440 = 3.1 (1.5), 10.9 (4.5), and 7.9 (3.9) μM, C471/496 = 14.3 (3.0), 1.7 (0.6), and 1.4 (0.5) μM. And for wavelength set 2, log K335/450 = 4.50 (0.03), 4.96 (0.27), and 5.22 (0.08), log K471/496 = 5.02 (0.04), 5.42 (0.32), and 5.71 (0.09), C335/450 = 8.8 (0.5), 21.9 (7.9), and 18.7 (0.3) μM, C471/496 = 21.0 (2.5), 7.17 (1.2), and 7.09 (0.3) μM. The ability of the 415/440 nm SyF transect to characterize the main excitation and emission maximum of FA at 335/440 nm is evaluated. Relatively low concentration values returned by the model for this transect (415/440 nm) suggest that it is not entirely illustrative of the maximum. The model predictive capability is verified at pH 6 with two fluorescing Cu2+ chelating organic compounds, L-tyrosine and salicylic acid. This test confirms that the model is capable of providing good estimates of equilibrium binding parameters from multi-wavelength measurements of a mixed ligand system.