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Fate and Effects of Pollutants

41. Grant, J., "Sensitivity of Benthic Community Respiration and Pri

mary Production to Changes in Temperature and Light." Mar. BioL

(W. Ger.), 90, 299 (1986). 42. Pulich, W., Jr., and Rabalais, S., "Primary Production Potential of

Blue-Green Algal Mats on Southern Texas [USA] Tidal Flats." Southwest. Nat, 31, 39 (1986).

43. Pringle, C, et al, "In situ Nutrient Assays of Periphyton Growth in a Lowland Costa Rican Stream." Hydrobiologia, 134,207 ( 1986).

44. Lowe, R. L., et al, "Periphyton Response to Nutrient Manipulations in Streams Draining Clearcut and Forested Watersheds." J. N. Am.

Benthol Soc, 5,221(1986). 45. Noel, D. S., et al, "Effects of Forest Clearcutting in New England

[USA] on Stream Macroinvertebrates and Periphyton." Environ.

Manage, 10,661(1986). 46. Mclntire, C. D., and Amspoker, M. C, "Effects of Sediment Prop

erties on Benthic Primary Production in the Columbia River Estuary [Washington, Oregon, USA]." Aquat Bot, 24, 249 (1986).

47. Cazaubon, A., and Loudiki, M., "Microdistribution of Epilithic Algae on the Stones of a Corsican [France] Stream; the Rizzanese." Ann.

Limmnol, 22, 3(1986). 48. Lobo, E., and Buselato-Toniolli, T. C, "Exposure Time on Artificial

Substrates for Periphyton Community Settlement in the Low Part of the Cai River, Rio Grande do Sul, Brazil." Rickia (Portuguese), 12,35(1985).

49. Jenkerson, C. G, and Hickman, M., "Interrelationship among the

Epipelon, Epiphyton, and Phytoplankton in a Eutrophic Lake." Int.

Revueles. Hydrobiol, 71, 577 (1986). 50. Grant, J., et al, "The Interaction between Benthic Diatom Film

and Sediment Transport." Estuarine, Coastal and Shelf Set, 23,

225(1986). 51. Baillie, P. W., "Oxyg?nation of Intertidal Estuarine Sediments by

Benthic Microalgal Photosynthesis." Estuarine, Coastal and Shelf Sei., 22, 143(1986).

52. Luttenton, M. C, and Rada, R. G, "Effects of Disturbance on Epi phytic Community Architecture." J. Phycol, 22, 230 (1986).

53. Rushforth, S. R., et al, "Algal Communities of Springs and Streams in the Mt. St. Helen's Region, Washington, U.S.A. Following the

May 1980 Eruption." / Phycol, 22, 129 (1986). 54. Robinson, C. T., and Minshall, G. W., "Effects of Disturbance Fre

quency on Stream Benthic Community Structure in Relation to

Canopy Cover and Season." / N. Am. Benthol. Soc, 5,237 (1986). 55. Cattaneo, A., and Kalff, J., "The Effect of Grazer Size Manipulation

on Periphyton Communities." Oecologia, 69, 612 (1986). 56. Howard, R. K., and Short, F. T., "Seagrass [Halodule wrightii]

Growth and Survivorship under the Influence of Epiphyte Grazers."

Aquat. Bot, 24, 287(1986). 57. Lodge, D. M., "Selective Grazing on Periphyton: A Determinant of

Freshwater Gastropod Microdistribution." Freshwater BioL, 16, 831

(1986). 58. Vaughan, C. C, "The Role of Periphyton Abundance and Quality

in the Microdistribution of a Stream Grazer, Helicopsyche borealis

(Tricoptera: Helicopsychidae)." Freshwater BioL, 16, 485 (1986). 59. Fuller, R. L., et al, "The Importance of Algae to Stream Inverte

brates." J. N. Am. Benthol. Soc, 5, 290 (1986). 60. Kamura, S., and Choonhabandit, S., "Algal Communities Within

Territories of the Damselfish Stegastes apicalis and the Effects of

Grazing by the Sea Urchin Diadema spp. in the Gulf of Thailand."

Galexia,5, 175(1986). 61. Liddell, W. D, and Ohlhorst, S. L., "Changes in Benthic Community

Composition Following the Mass Mortality of Diadema at Jamaica

[West Indies]." J. Exp. Mar. BioL Ecol. (Neth.), 95, 271 (1986). 62. Pratt, J. R., et al, "Effects of Seasonal Changes on Protozoans In

habiting Artificial Substrates in a Small Pond." Arch. Protistenkd.

(Ger.), 131,45(1986). 63. Baldock, B. M., "Peritrich Ciliates Epizoic on Larvae of Brachycen

trus subnubilus (Tricoptera): Importance in Relation to the Total Protozoan Population in Streams." Hydrobiologia, 131,125 (1986).

64. Smith, M. E., "Distribution Patterns and Seasonal Occurrence of

Rhabdostyla sp. (Peritricha: Epistylididae) on Dew nivea (Oligo chaeta: Naididae)." Am. Midi Nat, 116, 348 (1986).

65. Kyle, D. E., and Noblet, G. P., "Seasonal Distribution of Ther motolerant Free-Living Amoeba: I. Willard's Pond [South Carolina, USA]." / Protozool, 33, 422 (1986).

66. Strayer, D., and Likens, G. E., "An Energy Budget for the Zoobenthos of Mirror Lake, New Hampshire [USA]." Ecology, 67, 303 (1986).

67. Wallace, R. L., and Edmondson, W. T., "Mechanisms and Adaptive Significance of Substrate Selection by a Sessile Rotifer." Ecology, 67,314(1986).

68. Sarvala, J., "Patterns of Benthic Copepod Assemblages in an Oli

gotrophic Lake." Ann. Zool Fenn., 23, 101 (1986). 69. Lundalv, T., et al, "Long-Term Trends in Algal Dominated Rocky

Subtidal Communities on the Swedish West Coast-A Transitional

system?" Hydrobiologia, 142, 81 (1986). 70. Coull, B. C, "Long-Term Variability of Meiobenthos: Value, Syn

opsis, Hypothesis Generation and Predictive Modelling." Hydro biologia, 142, 21 \ (1986).

71. Herman, P. M. J., and Heip, C, "The Predictability of Biological Populations and Communities: An Example From the Meioben thos." Hydrobiologia, 142, 281 (1986).

Human health effects

assays Ronald C. Sims, Judith L. Sims, R. Ryan Dupont

CURRENT APPROACH Health risk assessment of individual environmental chemicals,

risks of drinking water supplies, methods for evaluating toxicity, and structure-activity relationships and response-surface meth

odology represent the focus for 1986. A report of the Steering Committee on Identification of Toxic and Potentially Toxic Chemicals for Consideration by the National Toxicology Pro gram found that, for the majority of substances they examined, data essential for human health hazard assessment were lacking.1

Milestone publications concerning human health effects aspects include works by Mortelmans et al,2 Busch et al,3 Hartman et

al,4 Rein,5 Gentile,6 Hodgson,7 and Tagashira and Omura.8

Public health aspects of hazardous waste disposal were addressed at a meeting of the Universities Associated for Research and Education in Pathology.9 Baram and Kenyon10 discussed several areas of the law that provide rights of access and duties to disclose risk with regard to chronic health and environmental hazards arising from exposure to chemical substances under "routine"

operations. The general areas of estimation of health risks and assessing

chemical hazards of environmental chemicals were addressed

by Kenaga,11 Cothern et al,12 Hoel et al,n and Ricci.14 The use

of risk assessment for making decisions for industrial chemicals already on the market was considered by Schultze and Mucke.15

Specific chemicals addressed using risk assessment included vinyl chloride,16 dioxin,17 and zinc, cadmium, and calcium.18 A special issue of The Science of the Total Environment was devoted to

June 1987 601


risk management in chemical safety.19 Validation methods for hazard assessment mathematical models were discussed by Burns.20

Assessment of drinking water for carcinogens and for exposure was discussed by Dixon et al.21 Ram et al.f22 and Munro and

Travis.23 The association of water contaminants with cardio

vascular disease and cancer was summarized by Craun.24 The

U. S. EPA Technical Support Document provided guidance for each step in the water quality-based toxics control process from

screening to compliance monitoring.25 Finally, chemical qualities of water that contribute to human health in a positive way were

discussed by Hopps and Feder.26 Analysis of data obtained from health effects assays were dis

cussed by Bois et al.27 for the Microtox test, and by Mitchell and Brice28 for the in vivo micronucleus test. A computer pro

gram in BASIC was designed by Abou-Setta et al.29 for deter mining probit and log-probit or logit correlation for toxicology. Clayson and Krewski30 examined three areas of difficulty with the concept of negativity in experimental carcinogenesis, in

cluding: the existence of chemicals that do not induce cancer, the melding of positive and negative assay results, and the pos

sibility of thresholds. A review of structure-activity relationships among mutagens

and carcinogens was presented by Frierson et al.31 The relation

ship between specific molecular connectivity indexes and tera

togenicity, carcinogenicity, and mutagenicity was discussed by Murakami and Fukami for carbamate pesticides32 and for chlo

rinated benzenes.33 The application of response-surface meth

odology to detect interactions of genotoxic agents in cultured mammalian cells was used by Wilson et al.34 for statistical design and analysis of experiments involving multiple variables.

Modeling approaches to health effects assays were addressed

specifically for neurotoxicology by Veronesi35 and for environ

mentally induced cancer using the multistage model of carci

nogenesis by Gaffney and Altshuler.36 Evaluation and compar ison of the National Toxicology Program battery of short-term tests with an alternate battery for 70 noncarcinogens were de

scribed by Ennever and Rosenkranz.37 The relationship between chemically induced patterns of turimorigenesis in rodents and of in vitro genetic toxicity was evaluated for 73 substances by Tennant et al3S A comparison of m vivo mammalian assay results

with in vitro results for predicting mutagens was presented by Natarajan and Obe.39 The effect of sex differences on toxic sus

ceptibility was summarized in a book by Calabrese.40 A compar ison of routes of administration of test chemicals in in vivo animal studies (intraperitoneal and gavage), and a case for the continued

use of the intraperitoneal route of exposure were presented by Shelby.41 A comparison of singular versus synergistic modulation

of lymphocyte mitogenesis by carcinogenic xenobiotics was conducted by McCabe and Nowak.42 A continuous liquid-liquid extraction system for concentrating trace organic chemicals in

the presence of humic materials was designed and evaluated by Baker and Suffet.43

Levin and Ames44 reported the development of two additional strains of Salmonella for detecting base changes, in addition to the strains presently used to detect base pair substitution mu

tations. The strains were used for classifying mutagens as to

their specificity in causing the six possible base changes (tran sitions and transversions).

Automation of microbial mutagenicity test systems by re peated optical density measurement of liquid cultures was dem

onstrated and discussed by Gocke and Schupbach.45 Automation was considered a promising way to reduce the manual work

load of a test laboratory.

TEST SYSTEMS

Recent biological test systems developed for identification of genetic toxicity included whole organism and cell assays iden tified in Table 1. Staining techniques were developed for eval uating the induction of aneuploidy in mammalian cells,65 mech anisms of chemically induced aneuploidy were discussed,66'67 and testing approaches were reported.68 However, differential

staining of chromosomes and spindle was not useful for iden

tifying the effect of cancer promoters on primary cultures of human fibroblasts.69

Comparisons of results using in vivo and in vitro assays were reported for cytogenic assays,70 and high performance liquid chromatography was used to confirm some differences between in vitro and in vivo metabolism of the polycyclic aromatic hy

drocarbon fluoranthene.71 Statistical analyses for in vitro cyto

genic assays using CHO cells were also summarized.72 Interlab

oratory testing was used to compare enhancement of Simian

Adenovirus SA7 transformation in Syrian Hamster Embryo Cells for environmental chemicals.73"75

Test systems using multiple genetic endpoints were used to evaluate the agents aminofluorene and dimethylnitrosamine.76 The endpoints included mutations at the sodium/potassium ATPase (ouabain resistance) and HGPRT loci, SCEs in Chinese hamster V79 cells, and mutation of Salmonella typhimurium strains TA98 and TA 100. Modification of the SOS Chromotest system, including automation and optimization of the method, increased the sensitivity of the test for progenotoxicity.77

A method for preliminary screening of polycyclic mutagens within 20 minutes was introduced by Tomoda et al.7* The method uses a biomimetic electrode composed of an oxygen electrode and a copper-phthalocyanine membrane.

ENVIRONMENTAL SAMPLES

Results of assays of drinking waters and surface waters are presented in Table 2, while results for municipal wastewaters, industrial wastes, and other environmental samples of concern

are presented in Table 3.

SPECIFIC CHEMICALS

Information concerning assays of individual organic chemicals or classes of chemicals and individual inorganic chemicals that have been evaluated for public health impacts has been presented in Tables 4 and 5, respectively.

SPECIAL ASPECTS OF HEALTH EFFECTS ASSAYS

The preparation of water soluble fractions of crude oils for toxicity assays was addressed.216 Conditions of mammalian cell

tissue cultures, including low pH and high osmotic levels, that may cause genotoxicity were addressed at a workshop held at

the 16th Annual Meeting of the Environmental Mutagen So ciety.217 Special aspects of conducting human health effects assays that were identified and evaluated are summarized in Table 6.

602 Journal WPCF, Volume 59, Number 6


Table 1?Miscellaneous cell and whole organism assays used for genotoxic evaluation of environmental chemicals.

Assay Endpoint Chemical(s) Reference

E. coli K-12 SOS chromotest

Soybean test system

Drosophila melanogaster

Metallothionein protein variants in rat liver

Marine fish S9?Salmonella

preincubation

Micronucleus test using newt

peripheral blood erythrocytes

Aspergillus nidulans Strain XD83

Toadfish kidney cells

Saccharomyces cerevisiae

Cell culture (human and mon

key growing cells)

Human lymphocyte SCE

Human IMR-90 fibroblasts

Salmonella typhimurium (TA 1538/TA 1978), E. coli K

12, E. coli WP2

Mouse liver tumors

Syrian hamster embryo/simian adenovirus SA7 (SHE/SA7) viral enhancement assay

Whole blood cultures (WBC) and plasma leukocyte cul tures (PLC)

Oyster toadfish and American eel peripheral lymphocyte test

Murine colonie tissue SCE

[3H]thymidine incorporation

Cell filamentation induced by DNA

damaging agents

Somatic mosaicism for detection of

genetic and/or chromosomal structural changes

Microsomal monooxygenase activity

Ethylation of nucleic acids as pro

mutagenic lesions

Mutagenicity

Cytogenetic damage, mutagenicity

Chromosome malsegregation, mi totic crossing-over, forward muta tion induction

Chromosomal aberrations and SCEs

Forward mutation, back nuclear frameshift and base-pair substitu tion mutation, nuclear intragenic and intergenic recombination, and mitochondrial forward point mutations and deletions

Aquatic toxicity

Sister chromatid exchanges

DNA-synthesis, aryl hydrocarbon hydroxylase activity

Bacterial DNA damage

Carcinogenicity

Transformation of primary SHE tar

get cells

SCE induction in human and pig cultures

SCE induction

SCE frequency

In vivo evaluation of [3H]thymidine incorporation into mouse organs

44 chemicals

EMS

2,3,7,8-TCDBF

Diethylnitrosamine

Seven genotoxic carcinogens

BaP, EMS, DES, ENNG

10 chemical carcinogens ("false negative" in bacterial

mutagenicity assays)

EMS, cyclophosphamide

Ultraviolet light

Cd, Cu, Zn

Mutagens

7,12-dimethylbenzanthracene

26 pesticides and pesticide degradation products

Mouse liver carcinogens

BaP

Ultraviolet light

Mitomycin C, EDB, dimethoate

1,2-dimethylhydrazine

1,2-dibromoethane, 1,2-di chloroethane

46

47

48

49

50

51

52

53

54

55

56

57

58

59

60

61

62

63

64

Table 2?Results of health effects assays of drinking waters and surface waters.

Type of sample Assay Comments Reference

Drinking water

Chlorinated drinking water Various mutagenicity assays

Review of drinking water and cancer mortality 79

Results of studies of mutagenic activity in chlorinated 80-86

drinking water, including identification and/or isola tion of mutagenic constituents

June 1987

(Table 2 Continued)

603


Table 2?(Continued)


Drinking water disinfectants and disinfection by-products

Acrylamide (ACR) in drinking wa ter (flocculant used in water

treatment)

Fluoride (potential drinking water

additive)

Drinking water contaminated with

4-chloro-3-methylphenol (CMP), a cutting oil

Water from Nishitakase River,

Kyoto City, Japan

Water from Torch Lake, Michigan

Organic river sediment pollutants

Extracts of blooms of blue-green

algae

Humic acids in water

Surface waters contaminated with phenols

Reproductive toxicity in rats and hepta-cellular DNA

repair

Lipid peroxidation in liver, brain, and intestine of

rats; DNA-repair in human fibroblasts and rat hepa tocytes

Ames and subacute toxicity in rats

Ames

Ames

Activity of mixed-function oxidases in rat liver

Toxicity to rat hepatocytes and to female mice

Mutagenicity

Permeation through hairless mouse skin

Symposium on health effects

ACR produced adverse reproductive effects in rats, but was not genotoxic in isolated rat hepatocytes

Results of studies of fluoride effects on lipid peroxi dation and mutagenic potential

Drinking water in Denmark contaminated with CMP,

probably from water pipes

Six different frameshift mutagens, all requiring meta bolic activation, were present

Lake water has high levels of dissolved copper from

copper mining activities

Evaluation of metabolic enzyme induction by sedi ment from mouth of Tama River in Japan, which receives domestic sewage and industrial waste

water

Hepatocyte-toxicity assay system potentially useful for screening for contamination by blue-green algae

Humic acid was not mutagenic and inhibited muta

genicity of benzo-[a]pyrene and 3-aminoanthra cene

Permeation dependent on pH of surface water

87

88-89

90-91

92

93

94

95

96

97

98

Table 3?Results of health effects assays of municipal wastewaters, industrial wastes and other environmental samples.


Municipal wastewaters

Tuskegee wastewater

Urinary mutagens in municipal sewage workers

Complex hazardous wastes

Paper mill effluent

Ames

Black bullhead fish

Ames

Modified version of TLC/ Salmonella assay

Induction of micronuclei in peripheral erythro

cytes of fish

Evaluation of mutagenicity at various

stages of conventional treatment

processes

Neoplasms developed on fish, al

though chemical analysis failed to indicate presence of mutagenic or

carcinogenic chemicals

Sewage workers had higher risk for uri

nary mutagens than water treatment workers

Evaluation of technique that involves direct application of Salmonella as

say to thin-layer chromatogram of 10 hazardous wastes (inorganic and

organic wastes in solid, semi-solid and liquid forms)

Evaluation of use of an aquatic verte brate for potential genotoxic effects

99

100

101

102

103



Table 3?(Continued)


Spent bleaching liquors from

sulphite and kraft pulps

Soils amended with two refinery wastes

Soils amended with two refinery wastes and two wood pre servative wastes

Petroleum-derived complex mixtures

Crude and refined oils

Used crankcase oils (UCO) from diesel and spark igni tion automobiles

Coal gasification wastes

Hydrotreatment of shale oil

Mutagenicity

Ames and Aspergillus methionine

Ames

Modified Ames

Mutagenicity using uni

cellular alga Chlamy domonas reinhardtii

Ames

Short-term bacterial mu

tagenicity

Ames

Mutagenicity from spent liquors from

sulphite pulps less than from kraft

pulps

Mutagenic potential of both wastes re duced by soil incorporation

Mutagenic potential of refinery wastes

generally reduced by soil incorpora tion but potential of wood preserva tive wastes at higher loading rates not reduced

Prescreening study recommended us

ing range of S9 concentration and

S9 preparations from different spe cies

Study of use of typical marine biota for determination of mutagenic po tential in aquatic environments

Health risks of UCO from diesel en

gines not significantly different of UCO from gasoline engines

Isolation and identification of muta

gens

Photomutagens and promutagens present in parent crude but only photomutagens present in hydro treated material

104

105

106

107

108

109

110-111

112

Table 4?Health effects assays of specific inorganic chemicals or classes of environmental concern.

Chemical Assay Comments Reference

Aliphatic epoxides

Aliphatic and polyhalogen ated carcinogens

Aromatic amines

Azapyrenes

Benzo(a)pyrene(BP) BP-diones

Furan analogues of BP and their 2-nitro d?riv?tes

Nitro-BP isomers

Benzyl chloride

Carbon tetrachloride

Chlordane

Chlorolefins

June 1987

Ames

Ames

Ames and anchorage indepen dent survival

Syrian hamster fibroblasts

Ames and SOS chromotest

Hepatocyte-mediated Salmonella

mutagenicity

rat fetal development; rat liver functions

Acute, subacute, and sub

chronic studies in rats

Ames

36 compounds comprising 6 subclasses tested

Mechanisms of carcinogenic actions

Investigation of induction of mixed function oxi dases and relationship to mutagenicity

Compounds tested were mutagenic; some were carcinogenic

Health effects assessment

BP-3,6-dione found to be mutagenic, cytotoxic, and to induce DNA damage

Responses in two test systems correlated; re

sponses to 2-nitro derivatives high; lower re

sponses with furan analogues

Hepatocytes capable of producing metabolites different from those of S9

Evidence of fetotoxicity and hepatomalfunction

Evidence of hepatic injury


Evidence of indirect mutagenic activity

113

114

115

116

117 118

119

120

121-122

123

124 125

(Table 4 Continued)

605


Table 4?(Continued)


Cyclophosphamide

DDT

Diazinon

Diazinon

1,2-dibromo-3-chloropro pane (DBCP)

DBCP

DBCP

DBCP

p-Dichlorobenzene

1,1-Dichloroethane

Dimethoate

Dinitrobenzene

Dioxins

Dioxin (TCDD) Dioxins (nitropolychlorinated

dibenzo-p-dioxins)

Ethylbenzene

Formaldehyde

Lindane

Malathion

4,4-Methylenedianiline

Monosodium methanear sonate

Naphthalene

M-Nitrobenzaldehyde

Mono-nitrobenzene deriva tives

4-Nitrobiphenyl

Nitrobiphenyls

Nitropyrene and derivatives

Cytogenetic and embryo-toxic ef fects in chick embryo

Calcium transport and thymidine uptake of bovine lymphocytes

Heart and skeletal muscle en

zyme activities

Alterations in peripheral blood

lymphocytes

Mouse spot test

Mouse-specific locus

Drinking water reproduction study in rats

Modified Ames

T?ratologie

Drosophila melanogaster ge netic damage

Reproductive performance, sperm quality, and reproduc tive organ histology in male rats

Ames

Chortophaga (grasshopper) em

bryos

RNA polymerase activity of all nuclei and transcription prod ucts in lymphocyte culture

Carcinogenicity

Hematology, growth, and repro duction of mice

Ames and DNA breakage in rat

hepatocytes

Ames and rec

Ames

Ames

Various

Teratogenicity associated with mutagenic activity

Study of relationship between chemically in duced leukemias and DDT exposure

Chronic low levels had little effect

Study of mutagenic potential

Demonstrated to be mutagenic in somatic cells

Negative results for induction of heritable gene mutations

No adverse reproductive effects demonstrated

Study of DBCP biotransformation and induction of acute extrahepatic toxicity

Teratogenicity not demonstrated


Slight increase in frequency of point mutations

Potent testicular toxicant

Symposium proceedings, including toxicologi cal, epidemiological, and risk aspects of hu man exposure

Health risk assessment approach

Newly identified class of compounds contains some bacterial mutagens


Induction of acentric chromosome fragments and chromosome stickiness

Drinking water criteria document

Decrease in polymerase activity and inhibition of RNA synthesis

Evidence of carcinogenicity

Decrease in reproductive capabilities; no effect on hematology or growth


Demonstration of weak genotoxic activity

Rec assay gave more positive results than Ames for 37 derivatives

Bladder tissues contained metabolic capability to activate carcinogenic potential

Study of relationship between mutagenic potential and chemical structure of nitro

biphenyls

Studies of mutagenicity and genotoxicity

126

127

128

129

130

131

132

133

134 135 136

137

138

139 140

141 142

143

144

145

146

147

148

149

150

151

152-158



Table 4?(Continued)


Dialkyl N-nitrosamines

N-Nitrosopropylamines

Oxygen free radicals

N-substituted phenanthrene 9,10-imines

Phenol

Phosphamidon

Pirimiphos-methyl

Polychlorinated biphenyls (PCBs)

Polycyclic aromatic hydro carbons

Solvents

Tetrachloroethylene

Thimet 10-G

Toluene

Trichlorfon

Vinylidene chloride

Xylenes

Hamster hepatocyte V79 cell mediated

Ames liquid incubation assay

E. coli DNA-repair

Ames and Chinese hamster V79 cells

Histopathological and biochemi cal changes in mouse liver

Micronucleus and sperm mor

phology

Serum analytes

Mixed-function oxidase (MFO) activity in various mouse lym phoid tissues

Ames, Mycobacterium, micronu

clei formation in mice, and SCE in human lymphocytes

Teratogenicity

Study of relationships between structure, meta bolic activation, and mechanisms of muta

genic activity

Study of organ-specific metabolic activation

Mutagenic and cytotoxic effects

Mutagenic effects


Study of effects on liver

Study of mutagenic properties

Study of relationship between PCBs and serum

analytes to assess health effects

Induction of MFO activity varied with type of

lymphoid tissue

Review of toxicology

Carcinogenicity assessment

Insecticide not mutagenic in the 4 test systems

Drinking water criteria document

Evidence of teratogenicity and fetotoxicity at

high dose levels

Review of mutagenicity

Review of toxicological information and health effects

159

160 161 162

163-164

165

166

167

168

169 170 171

172 173

174 175-176

Table 5?Health effects assays of specific inorganic chemicals of environmental concern.


Asbestos

Arsenic

Arsenic trioxide and sodium arsenate

Sodium arsenite

Beryllium

Cadmium

Cadmium

Cadmium

Cadmium

Human excision DNA repair

Chinese hamster ovary and

human skin fibroblasts

Pathophysiological

Chinese hamster ovary

Male rat reproduction func

tions

Chronic health effects


Evidence of inhibition of DNA repair

Enhancement of clastogenicity and

mutagenicity of DNA cross-linking agents


Review of toxicology of cadmium

Studies of effects on cardiovascular

system, erythropoiesis, and ner vous system; lifetime effects of Itai-itai disease

Effects on cell division and chromo somes

Study of response characteristics in

terms of age and dose

177 178 179

180

181 182

183-187

188

189

June 1987

(Table 5 Continued)

607


Table 5?(Continued)


Chromium

Cobalt

Lead

Lead

Mercury

Mercury

Mercuric chloride

Methylmercury

Methylmercury

Nickel and nickel salts

Organometals

Sodium selenite

Dioctyltin dichloride

Vanadium

Various mutagenic and patho

physiological

Ames

Pathophysiological

Hematology in chickens

Pathophysiological

Blood pressure in sponta neously hypertensive rats

Glutathione peroxidase activity

Mutagenicity

Neurotoxicity

Peripheral leukocytes in fe male mice

Mouse immune responsive ness to self and heterolo

gous cell membrane com

ponents

Rat intestinal absorption of calcium

Studies of effects of various chro mium compounds

Increased mutagenicity due to for mation of complexes between co

balt cation and organic chemicals

Review of health hazards

Studies of effects on rat tissues and mouse reproduction


Study of interaction of mercury and water deprivation

Studies of effects on liver and kidney functions

Studies of toxicity, including sex dif ferences

Inhibition of liver activity

Studies of mutagenic potential and mechanisms of action

Review of macrophysiological inves

tigations

Decreased levels following treatment

Suppression of immunocompetence

Study of calcium transport into duo denum

190-196

197

198-199

200-201

202 203

204-205

206-207

208 209-211

212

213

214

215

Table 6?Special aspects of human health effects assays.

Assay Comments Reference

Salmonella typhimurium/hepaiocyte

Salmonella typhimurium

Salmonella typhimurium

Salmonella typhimurium/erythrocyte

Salmonella typhimurium forward mutation assay

Cultured C3H/10T1/2 cells

CHO cells

In vitro mutagenicity assays

L5178Y Mouse lymphoma

Effect of source of hepatocytes (monkey and man) evaluated

Effect of toadfish hepatic postmitochondrial fractions on activation and deactivation of promutagens

Effect of human hepatocytes versus human liver S9 on mutagenic activity. Results also compared with results obtained with rat liver preparations

Comparison of washed red blood cells of mice and S9 fraction from Aro

clor-1254-induced rats for activation of 2-aminofluorene (2-AF)

Permits measurement of cell survival following mutagen treatment by plat

ing the culture on specially supplemented plates at same concentration

used to measure mutant yield

Development of methods to assess and to improve the efficiency of deliv

ering a petroleum sample (complex hydrophobic mixture) to assay cells

for genotoxicity studies

Differences in effects of types of sera (newborn versus fetal bovine), amounts and batches of sera

Behavior of benzene and formaldehyde in culture medium as affected by fetal calf serum greatly affects testing protocol

Effect of agar on results were characterized on basis of agar suppliers

(agar type)

218 219

220

221

222

223

224

225

226



Table 6?(Continued)

Assay Comments Reference

Animal cell DNA polymerase fidelity Provides information on DNA polymerases from animal cells to comple- 227 ment DNA polymerase from other eukaryotic organisms

Chinese hamster V79 spheroids Model of mutagen "penetrability" by providing several layers of cells 228

growing with tissue-like packing

Micronucleus test Sex differences suggest that use of male mice is sufficient for general 229

screening of clastogens

SCE Comparison of two external metabolizing systems, Aroclor-induced rat liv- 230 ers and hepatocytes, used for activation of human lymphocytes and V79 cells

Ronald C. Sims, Judith L. Sims, and R. Ryan Dupont are with Utah State University. Correspondence should be addressed to Ronald C. Sims, Utah Water Research Laboratory, Utah State

University, Logan, UT 84322-8200.

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173. Courtney, K. D., et al, "Assessment of Teratogenic Potential of Trichlorfon in Mice and Rats." / Environ. Sei. Health, B(21), 207

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Effects of chemicals on microorganisms

John D. Walker

"Toxicity Testing Using Microorganisms," a two-volume text was published.1 The first volume contains a brief overview of screening procedures,2 and detailed chapters on several screening

procedures including bacterial growth,3 biochemical tests,4 bio luminescence assays,5 respirometric techniques,6 and microcal

orimetric studies.7 The second volume contains a brief overview2 and chapters on factors that affect microbial assays,8 toxicity screening for fungi and yeasts,9 phytoplankton,10 wastewater,11 soil,12 biod?gradation tests,13 and a perspective on microbial toxicity testing.14

BACTERIAL TOXICITY TESTS

Bioluminescence. P. phosphoreum was used in the Microtox?

test to: assess the toxicity of zinc and pentachlorophenol (PCP); determine that interactions between zinc and PCP were additive; and illustrate that use of the isobolographic method to estimate chemical interactions may lead to erroneous conclusions about

those interactions.15 Studies were conducted to determine the influence of exposure time, test temperature, pH, salinity, and alkalinity on the toxicity of cadmium, zinc, PCP, and benzene in the Microtox? test. Exposure time increased the toxicity of cadmium and zinc, but decreased the toxicity of benzene; in general, chemicals were more toxic at 20?C than at 15?C, at pH 6 rather than pH greater than 6, at 1.5% NaCI concentrations, and in soft water (Tables 2, 4, 8).16 P. phosphoreum was also used in a LBK-Wallach luminometer to measure differences in efficacy levels for a biocide-inhibitor stored for 3 months in steel containers and glass containers; storing the biocide-inhibitor in steel containers significantly reduced efficacy.17

Microcalorimetry. Fast-flow microcalorimetry was used to

determine the toxicity of copper, mercury, zinc, phenol, sodium

dodecyl sulfate (SDS), and cetyl trimethylammonium bromide (CTAB) to mixed bacterial cultures; results were compared with the Microtox? test and with acute toxicity tests conducted with the aquatic invertebrate, Daphnia magna and the freshwater alga, Selenastrum capricornutum (Tables 4, 5, 6, 8, 9).18

Resazurin reduction. Details of a 20- to 30-minute resazurin

reduction screening method using Bacillus cereus to measure

toxicity of soluble and insoluble chemicals in waters or to assess toxicity-structure relationships were recently published.19 The

procedure was used to determine the effects of carrier solvents,

dimethylsulfoxide (DMSO), methanol, ethanol, and acetone on the toxicity of PCP and seven chlorobenzenes (Tables 2,4). The authors concluded that toxic responses were independent of car

rier solvent, but for chemicals with low water solubility, DMSO should facilitate determination of chemical toxicity.20

Tetrazolium dye reduction. A 30-minute INT-dehydrogenase

assay (DIDHA) using the spent medium of Pseudomonas al caligenes was developed to directly measure (without Formazan

extraction and centrifugation) the toxicity of metals, phenol,


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