Effects of Polychlorinated Biphenyl Compounds and Proposed PCB-Replacement Products on Embryo-Larval Stages of Fish and Amphibians

Static renewal bioassays were performed with four polychlorinated biphenyls, including Capacitor 21 and Aroclors 1016, 1242, and 1254. Each compound was used to treat embryo-larval stages of the rainbow trout (Salmo gairdneri), channel catfish (Ictalurus punctatus), goldfish (Carassius auratus), redear sunfish (Lepomis microlophus), leopard frog (Rana pipiens), Fowler\u27s toad (Bufo fowleri), and American toad (Bufo americanus). Developmental stages of the trout were the most sensitive, with LC50 \u27s at 4 days posthatching of 0.3, 1.0, 1.1, and 1.6 μg/l for Aroclors 1254, 1242, 1016, and Capacitor 21, respectively. LC1\u27s ranged between 0.009-0.011 μg/l. Fowler\u27s toad was the most tolerant species, and LC50 \u27s varied from 3.7-28.0 μg/l. Toxicity of the PCB\u27s generally increased with percent chlorine substitution. Bioassays also were conducted with dioctylphthalate (D0P), diisononylphthalate, and Dow Corning 561 silicone, three products proposed as replacements for PCB\u27s. Embryos and larvae of the channel catfish, redear sunfish, leopard frog, and Fowler\u27s toad were exposed through 4 days posthatching. Catfish was the most sensitive species, and sunfish was the most resistant. Based on LC50 values, replacement products were found to be 2-5 orders of magnitude less toxic than PCB\u27s. Continuous flow embryo-larval bioassays were performed on Capacitor 21 and DOP. Test species included redear sunfish, rainbow trout, and largemouth bass (Micropterus salmoides). Paralleling results from static renewal tests, flow-through data reflected a vast difference in toxicity between PCB\u27s and potential replacements, and good data correlation was achieved between the two bioassay procedures

Sensitivity of Vertebrate Embryos to Heavy Metals as a Criterion of Water Quality: Phase III: Use of Fish and Amphibian Eggs as Bioindicator Organisms for Evaluating Water Quality

Fish and amphibian eggs, embryos and early posthatched (larval) stages were evaluated as bioindicator organisms with which to monitor the quality of natural water resources. Eggs of 9 species were cultured in water collected from each of 11 Inner Bluegrass rivers and streams. The latter were chosen to represent water sources varying in quality from extremely poor to good. Selection was based on the sources and magnitude of pollution, and the diversity and density of piscine populations. Cultures were maintained in vitro, using 12-hour changes of water. Averaging data for all 9 animal species, egg hatchability (embryonic survival) ranged from 0% for the most contaminated water to 94% for uncontaminated water found to support a healthy aquatic biota. Eggs of the squirrel treefrog and gray treefrog were the most sensitive of 5 amphibian species tested, and of 4 fish species, rainbow trout eggs proved most susceptible to water contaminants

Sensitivity of Vertebrate Embryos to Heavy Metals as a Criterion of Water Quality, Phase I

Avian, amphibian and fish embryos were given continuous treatment with inorganic mercury, methyl mercury, cadmium and lead, to determine the sensitivity of embryogenesis to metallic poisoning. All metals produced substantial degrees of lethality and/or gross anatomical anomalies at 10 ppb or less. Treatment with inorganic mercury at 10 ppb produced 100% kill of frog embryos. Chick and rainbow trout embryos suffered 10-20% lethality when exposed to 1 ppb of either inorganic or methyl mercury. Lead and cadmium at 1 ppb produced 24-32% lethality in chick embryos. No significant differences were observed in the embryopathic effects of inorganic or methyl mercury. Concerning the toxic effects of mercury, cadmium and lead, the embryonic stage appears to constitute the critical sensitive link in the vertebrate life cycle. The reproductive potential of vertebrate populations may be severely restricted (e.g., embryonic mortality) by such pollutants at trace levels which may not prove hazardous to adult animals, and environmental standards based on tolerance levels for adult animals may not provide adequate protection for sensitive developmental stages

Coexpression of Kit and the receptors for erythropoietin, interleukin 6 and GM-CSF on hemopoietic cells

The detection of functional growth factor (GF) receptors on subpopulations of hemopoietic cells may provide a further dissection of immature cell subsets. Since little information is available about coexpression of different GF receptors at the level of single hemopoietic cells, we studied the feasibility of simultaneous cell staining with a combination of biotin- and digoxigenin-labeled GFs for flow cytometric detection of functional receptors. Using this methodology, coexpression of Kit and receptors for erythropoietin (EPO), interleukin 6 (IL-6), and GM-CSF on hemopoietic cells was studied by triple-staining of rhesus monkey bone marrow (BM) cells with labeled GFs and antibodies against other cell surface markers. Most of the immature, CD34+2 cells were Kit+ but did not display detectable levels of EPO-receptors (EPO-Rs) or GM-CSF-R. Approximately 60% of these CD34+2/Kit+ cells coexpressed the IL-6-R, demonstrating that immature cells are heterogeneous with respect to IL-6-R expression. Maturation of monomyeloid progenitors, as demonstrated by decreasing CD34 and increasing CD11b expression, is accompanied by a decline of Kit and an increase in GM-CSF-R expression in such a way that Kit+/GM-CSF-R+ cells are hardly detectable. IL-6-R expression is maintained or even increased during monomyeloid differentiation. IL-6-R and GM-CSF-R were not identified on most CD71+2 cells, which indicated that these receptors are probably not expressed during erythroid differentiation. Together with previous results, our data show that both Kit and CD71 are upregulated with erythroid commitment of immature progenitors. Upon further differentiation, Kit+/EPO-R-cells lose CD34 and acquire EPO-R. Maturing erythroid cells eventually lose CD71 and Kit expression but retain the EPO-R. In conclusion, this approach enables further characterization of the specificity of GFs for different bone marrow subpopulations. Apart from insight into the differentiation stages on which individual GFs may act, information about receptor coexpression may be used to identify individual cells that can respond to multiple GFs, and allows for further characterization of the regulation of lineage-specific differentiation

Sensitivity of Vertebrate Embryos to Heavy Metals as a Criterion of Water Quality, Phase II: Bioassay Procedures Using Developmental Stages as Test Organisms

Chick, amphibian, and fish embryos were evaluated as bioassay and bioindicator organisms. Test procedures were developed by which embryonic stages may be used 1) in bioassay systems to evaluate the toxicity of particular metallic or metal-containing trace contaminants, and 20 as bioindicators to monitor the quality of natural water resources. A bioassay technique was devised in which metallic toxicants were administered to chick embryos by needle tract injection into the yolk sac. This provided more uniform distribution of test metals throughout the yolk mass than can be obtained by conventional yolk sac injection methods, and gave more sensitivity and uniformity of test results. Metals such as arsenic, cadmium, mercury, lead and zinc are easily detectable at a level of 1 ppb. An in vitroculture technique was developed by which embryos of aquatic vertebrates may be maintained for bioassay and bioindicator purposes. Five test species were identified, suitable synthetic culture water was formulated, and culture monitoring procedures were determined. Most toxic metals (e.g., mercury) may be detected at 1ppb or less with the use of more sensitive embryonic species (e.g., trout). Early cleavage stages of the leopard frog (Rana pipiens) proved more sensitive to cadmium than older embryos, similar to results obtained in Phase I with mercury treatment of frog embryos. Early developmental stages, therefore, have proven especially important for use in bioassay and bioindicator systems

Toxicological Studies on Aquatic Contaminants Originating from Coal Production and Utilization: The Induction of Tolerance to Silver in Laboratory Populations of Fish and the Chronic Toxicity of Nickel to Fish Early Life Stages

Aquatic toxicity studies were performed on two important coal-derived contaminants, silver and nickel. Silver was investigated with regard to metal-induced tolerance in laboratory populations of the fathead minnow (Pimephales promelas). Fish were exposed to acute silver concentrations following acclimation to sublethal exposures of this metal. Based on median lethal times (LT50), animals which had received 14 days prior exposure to 1.5 and 15 μg Ag/L were three to four times more resistant to silver than were previously unexposed organisms. This metal-induced resistance was not a sustained response. After organisms which had been acclimated to 15 μg/L had been transferred to clean water for two weeks, LT50 values determined with these animals were statistically indistinguishable from those calculated with non-acclimated control fish. With respect to nickel, a 32-day continuous-flow test was performed with the fathead minnow. Nickel was administered in duplicate at six exposure concentrations ranging from 0.038 to 0.733 mg/Lin medium-hard water (100 mg CaC03/L). Animal test responses included mortality, teratogenesis, and growth. Based on frequencies of mortality after 32 days of continuous exposure, significant effects were recorded at a nickel concentration of 0.120 mg/L, and the no observed effect concentration (NOEC) was determined to be 0.057 mg Ni/L

Geometric realizations of generalized algebraic curvature operators

We study the 8 natural GL equivariant geometric realization questions for the space of generalized algebraic curvature tensors. All but one of them is solvable; a non-zero projectively flat Ricci antisymmetric generalized algebraic curvature is not geometrically realizable by a projectively flat Ricci antisymmetric torsion free connection

Embryopathic Effects of Waterborne and Sediment-Accumulated Cadmium, Mercury and Zinc on Reproduction and Survival of Fish and Amphibian Populations in Kentucky

Fish and amphibian egg cultures were used to determine the embryopathic effects of cadmium, mercury, and zinc released from natural and metal-enriched sediments, and to develop egg culture bioassay procedures suitable for monitoring bottom sediments for hazardous contaminants. Eggs of the narrow-mouthed toad (Gastrophryne carolinensis), goldfish (Carassius auratus), and rainbow trout (Salmo gairdnei) were cultured in contaminant-free water added to natural and metal-enriched sediments. Exposure was initiated after fertilization (toad, goldfish) or at 10 days prehatching (trout) and maintained continuously through 4-10 days posthatching. Sediments were enriched with 0.1-100 ppm cadmium and mercury and 1.0-1000 ppm zinc. Natural control sediments contained average concentrations of 0.052 ppm mercury, 1.0 ppm cadmium, and 108.2 ppm zinc. Substantial frequencies of mortality and teratogenesis occurred for all 3 animal species when eggs were cultured over natural elements further enriched with as little as 0.1-1.0 ppm cadmium or mercury and 1-10ppm zinc. Survival of trout embryos and alevins closely paralleled sediment test concentrations. The sediment TL50 concentrations for trout stages cultured from 10 days prehatching through 10 days posthatching were approximately 1ppm for mercury, 2.15 ppm for cadmium, and 210.6 ppm for zinc. Sediment metals were substantially more lethal to eggs and embryos than to free-living larvae or fry

The Aquatic Toxicity of Organic Compounds to Embryo-Larval Stages of Fish and Amphibians

Aquatic toxicity tests were conducted on 11 organic compounds considered hazardous to water resources. The toxicity of each compound was evaluated using embryo-larval stages of two to eight fish and amphibian species. Exposure was initiated at fertilization and maintained through 4 days posthatching. The animal test species exhibited varying degrees of sensitivity to the selected toxicants. Combined frequencies for mortality and teratogenesis at 4 days posthatching gave LC50 ranges of 3.66 to 8.25 mg/L for benzene, 1.16 to 22.42 mg/L for carbon tetrachloride, 0.11 to 1.20 mg/L for chlorobenzene, 2.03 to \u3e 68 mg/L for chloroform, 3.01 to 5.56 mg/L for 1,2-dichlorobenzene, 2.54 to .34 mg/L for 1,2-dichloroethane, 13.16 to \u3e 48 mg/L for methylene chloride, 0.002 to 0.64 mg/L for nitrobenzene, 0.04 to .32 mg/L for phenol, 0.02 to 0.85 mg/L for toluene, and 3.53 to 3.77 mg/L for m-xylene. The species which exhibited the greatest susceptibility to organic compounds were the rainbow trout, Rana pipiens, and Rana temporaria. The more sensitive amphibian species generally were those which normaly are restricted to aquatic or moist terrestrial habitats, whereas the more tolerant amphibians included those semi-aquatic and terrestrial species which appear to be more broadly adapted ecologically. Of the 11 test compounds, nitrobenzene, toluene, chlorobenzene, and phenol were the most toxic. The least toxic organics included dichloroethane and methylene chloride. For three chlorinated alkanes, including methylene chloride (CH2Cl2), chloroform (CHCl3), and carbon tetrachloride (CCl4), toxicity was found to 1ncrease with the degree of chlorination. Concerning several aromatic hydrocarbons, benzene always was found to be less toxic than its monosubstituted analogs. Toxicity of the 11 compounds was further evaluated by calculating toxicant concentrations which produced embryo-larval mortality and/or teratogenesis at frequencies of 10% (LC10) and 1% (LC1). The LC values, used to estimate toxicity thresholds, ranged from \u3c 0.l for nitrobenzene to 69.9 μg/L for methylene chloride. A limited number of toxicity tests were performed to determine whether embryo-larval bioassays are suitable to assess effects of transitory chemical exposures, such as those resulting from intermittent discharges or accidental spills of chemicals into water resources. Results indicated that Rana pipiens embryos were sufficiently sensitive to quantify effects produced by short-term exposures to chloroform. Animals tested during the earliest embryonic stage appeared to be less tolerant than organisms exposed later in development