ảnh hưởng của kln

ENVIRONMENTAL RESEARCH 37, 253-286 (1985)
REVIEW
The Mediation of Mutagenicity and Clastogenicity of Heavy
Metals by Physicochemical Factors
H.BABIcH,M. A. DEVANAS,ANDG.STOTZKY
Laboratory of Microbial Ecology, Department of Biology, New York Uni~~ersity, 1009 Main
Building. Washington Square, Nebt, York. Nen* York 10003
Received January 24, 1984
Heavy metals are an important class of environmental hazards. and as the use of heavy
metals in industry continues to increase, larger segments of the biota. including human
beings, will be exposed to increasing levels of these toxicants. As many heavy metals are
mutagenic and clastogenic, they cause teratogenic and/or carcinogenic effects. Studies with
microbes and representatives of the aquatic biota have shown that the toxicity of heavy
metals is mediated by the physicochemical characteristics of natural environments. A few
studies have also indicated that such abiotic factors (e.g.. pH. chelating agents. inorganic
anionic and cationic composition) mediate the mutagenicity and clastogenicity of heavy
metals. These studies indicate that the physicochemical characteristics of natural environ-
ments may also potentiate or attenuate the mutagenicity and clastogenicity of heavy metals
to the indigenous biota. Furthermore, studies with laboratory animals have shown that the
acute and chronic toxicity, including the teratogenicity and carcinogenicity, of heavy metals
is mediated by physicochemical factors. A similar dependence of the mutagenicity and
clastogenicity of heavy metals by the physicochemical characteristics unique to specific
body fluids and tissues may explain the association of specific heavy metal-induced tumors
with specific tissues. There is an apparent need to develop genotoxicity tests that incor-
porate into their procedures the mediating influence of physicochemical factors (PH. for
example), as the use of only standardized procedures may hinder the detection of heavy
metal, as well as of organic, genotoxins whose mutagenicity or clastogenicity is altered by
conditions other than those used in the standardized assay. thereby producing false negative
results. 0 198 Academic Press. Inc.
INTRODUCTION
Heavy metals are ubiquitous in the biosphere, where they occur as part of the
natural background of chemicals to which human beings and the biota are ex-
posed. However, industrial uses of metals and other industrial and domestic pro-
cesses (e.g., burning of fossil fuels, incineration of wastes, automobile exhausts,
smelting processes, and the use of sewage sludge as landfill material and fertilizer)
have introduced substantial amounts of potentially toxic heavy metals into the
atmosphere and into aquatic and terrestrial environments, thereby contaminating
food chains important to human beings. Consequently, such anthropogenic emis-
sions have elevated the body burden of heavy metals. For example, an increase
in the total body burden of cadmium (Cd) by a factor of five and in the kidneys
(i.e., the “critical organ”) by a factor of fifty has been noted since the first part
of this century (Drasch, 1983). Furthermore, as some metals adversely affect the
253
0013-9351/85 $3.00
Copyright D 1985 by Academic Press. Inc.
All rights of reproduction in any form reserved.
254 BABICH, DEVANAS, AND STOTZKY
genome, there is concern about the potential detrimental genetic effects of in-
creased exposure to such genotoxic agents: (1) an increase in the mutation rate
or in the incidence of chromosomal aberrations in germ cells caused by exposure
to heavy metals may hinder reproductive success (e.g., by inducing spontaneous
abortion) and/or increase the frequency of genetic disease in future generations,
and (2) mutations or chromosomal aberrations in somatic cells caused by heavy
metal exposure may lead to an increased incidence of cancer in the present gen-
eration (Hoffmann, 1982).
There is a growing recognition that heavy metals are an important class of
environmental and occupational genotoxins. There are several reviews of metal-
induced carcinogenesis (e.g., Sunderman, 1978, 1979; Flessel, 1979; Furst, 1981;
Jennette, 1981; Nordberg and Andersen, 1981; Vainio and Sorsa, 1981), and some
papers have evaluated the mutagenicity, clastogenicity, teratogenicity, and/or car-
cinogenicity of heavy metals, including Cd (Degraeve, 1981), chromium (Cr)
(Leonard and Lauwerys, 1980; Levis and Bianchi, 1982), mercury (Hg) (Leonard
et al., 1983), nickel (Ni) (Leonard et al., 1981), and lead (Pb) (Gerber et al., 1980).
Based on current evidence, at least ten metals can be considered carcinogenic,
with some compounds of arsenic (As), beryllium (Be), Cd, Cr, and Ni being
implicated as human carcinogens and some compounds of Be, Cd, cobalt (Co),
Cr, iron (Fe), manganese (Mn), Ni, titanium (Ti), and zinc (Zn) being implicated
in causing cancer in laboratory animals (Flessel, 1979; Sunderman, 1979; Man-
cuso, 1980; Wagoner et al., 1980). The genotoxic effects of some heavy metals
have been demonstrated: lesions in DNA (e.g., strand breaks and cross-links),
gene mutations, chromosomal aberrations, transformation of mammalian cells,
increases in the frequency of sister-chromatid exchanges (Kazantzis and Lilly,
1979; Heck and Costa, 1982; Christie and Costa, 1983; Tsapakos and Wetterhahn,
1983), and adverse effects on spindle formation, which has the potential to induce
aneuploidy (Onfelt and Ramel, 1979).
Several studies have indicated that environmental and occupational exposure
to heavy metals is a genetic hazard to human beings and the general biota. For
example, certain types of cancer appear to be more frequent among workers in
Ni refineries (lung, nasal sinus, larynx, and kidney carcinomas), Cd refineries
(lung carcinomas), and Cr refineries and among chrome platers and chromate
pigment workers (lung carcinomas) than in the population as a whole (Sunderman,
1979). An increased frequency of spontaneous abortions was noted in families of
workers occupationally exposed to Cr (Levis and Bianchi, 1982), and an increased
incidence of chromosomal aberrations occurred in individuals occupationally ex-
posed to Cd (deKnudt and Leonard, 1975), Cr (Sarto et al., 1982), Ni (Leonard
et al., 1981), and Pb (deKnudt et al., 1973, 1977).
Fish from a river in northern Illinois, which was polluted with As, Hg, Pb, toluene.
crude oil, gasoline, benzanthracene, chlorinated hydrocarbons, phosphates, sul-
fates, and coliform bacteria, exhibited an increased frequency of tumors when
compared with comparable species from nonpolluted Canadian waters (Brown et
al., 1973). Water samples from the Rhine River in Europe, which is heavily con-
taminated with organic pollutants, including polynuclear aromatic hydrocarbons
(Alink et al., 1980), and with heavy metals (Dissanayake et al., 1983), exhibited
mutagenic properties (van Kreijl et al., 1980). An increased incidence of sister-
METAL GENOTOXICITY: ABIOTIC FACTORS 255
chromatid exchanges and other chromosomal aberrations occurred in the gills
and testes of Umbra pygmaea maintained in Rhine River water (Prein et al.,
1978; Alink et al., 1980; Hooftman and Vink, 1981). Chromosomal aberrations
were induced in the gills of BoleophthaEmus dussumieri maintained in water
amended with W+ or phenyl Hg+ (Krishnaja and Rege, 1982). An increased
frequency of pollen abortion and mutations occurred in Zea rnays and Trades-
cantia? respectively, grown on soils in the vicinity of a Pb smelting plant emitting
large quantities of Cd, copper (Cu), Pb, and Zn (Lower et al., 1978).
Studies on the toxicity of heavy metals that have used microbes (Table l),
representatives of the aquatic biota (Table 2), or laboratory animals (Table 3) have
clearly demonstrated that the toxicity is influenced by environmental factors,
such as pH, the inorganic anionic and cationic composition, and the type and
concentration of chelating agents. However, there have been few comparable
studies on the effects of environmental factors on the genotoxicity of heavy
metals, even though studies with organic pollutants have indicated that some
environmental factors, pH in particular (Singh and Kaul, 1978), mediate muta-
genicity and clastogenicity. For example, the mutagenicity of N-nitrosodieth-
ylamine. N-nitrosodi-rz-butylamine, and N-nitrosomorpholine was greatest at pH
5.2, intermediate at pH 6.2, and least at pH 7.2 (Negishi and Hayatsu, 1980); the
mutagenicity of N-methyl-N’-nitro-N-nitrosoguanidine was greater at pH 6 than
at pH 7 or 8 (Adelberg et al., 1965); the mutagenicity of diethylnitrosamine was
greater at pH 5.6 than at pH 7.4 (Leonardo and Peak, 1982); decreasing the pH
from 8 to 6 increased the mutagenicity of ethyl nitrosourea (deKok et al., 1983);
and, conversely, increasing the pH from 5 to 9 increased the mutagenicity of the
fungicide, captan (N-(trichloromethylthio)-4-cyclohexene-l,2-dicarboximide)
(Bridges et al., 1972). The pH also affects the mutagenicity of some inorganic
compounds, including nitrite (Vielmetter and Schuster, 1960) and sodium azide
(deFlora and Boido, 1980).
The paper will discuss the potential of abiotic environmental factors to mediate
the genotoxicity of heavy metals. Few studies have investigated this question,
and those that have have indicated an interaction between abiotic factors (e.g.,
pH, inorganic anions and cations, organics) and heavy metal genotoxicity. It has
been suggested that the lack of consistency in microbial mutagenicity and tissue
culture clastogenicity assays for detecting the genotoxicity of heavy metals may
be due to the complexation of the heavy metals by the constituents of the assay
media, resulting in detoxification of the metals (McCann et al., 1975). Assays for
mutagenicity and clastogenicity that are so standardized that they employ essen-
tially only one set of environmental conditions might underestimate the potential
genotoxicity of chemicals whose adverse effect may be manifested only under
environmental conditions different from those used in the assays. Consequently,
the need to develop genotoxicity testing procedures for heavy metals that rec-
ognize and incorporate the mediating influence of physicochemical environmental
factors will also be discussed.
MECHANISMS OF THE GENOTOXIC EFFECTS OF HEAVY METALS
Heavy metals appear to function differently from other mutagens and carcin-
ogens, although this may only be apparent because so little is known about the
TABLE 1
INFLUENCE OF SOME PHYSICOCHEMICAL FACTORS ON THE TOXICITY OF HEAVY
METALS TO MICROORGANISMS
Physicochemical
factor Comments and references
PH Increasing the pH from acidic to alkaline levels reduced the toxicity of

Ni to bacteria, an actinomycete, a yeast, and lilamentous fungi (Babich
and Stotzky, 1982a, 1983a and d).
Increasing the pH from acidic to alkaline levels potentiated the toxicity
of Zn to filamentous fungi (Babich and Stotzky. 1983a) and a lilamen-
tous alga (Hargreaves and Whitton, 1976) but reduced that to a uni-
cellular alga (Rai er al., 1981).
Increasing the pH from acidic to alkaline levels reduced the toxicity of
Cd to a cyanobacterium (Singh and Pandey. 1981) but increased that
to bacteria and fungi (Babich and Stotzky, 1977a; Korkeala and Pek-
kanen, 1978); the effect of pH on the toxicity of Cd to filamentous
fungi was dependent on the composition of the assay medium (Babich
and Stotzky, 1983a).
Inorganic cations Mg reduced the toxicity of Ni to bacteria, a yeast (Abelson and Aldous,
1950), and filamentous fungi (Babich and Stotzky. 198lb, 1982d. 1983~)
and of Cd to a bacterium (Abelson and Aldous, 1950). a lilamentous
fungus (Laborey and Lavollay, 1977), and a tilamentous alga (Say and
Whitton, 1977).
Ca reduced the toxicity of Cd to an alga (Gipps and Coller, 1982) and of
Zn to a cyanobacterium (Shehata and Whitton, 1982) and algae
(Harding and Whitton. 1977; Rai et al., 1981).
Inorganic anions Increasing the chlorinity reduced the toxicity of Cd to tilamentous fungi
(Babich and Stotzky, 1982~) and of Hg to bacteria and coliphages (Ba-
bich and Stotzky. 1979b).
S’- reduced the toxicity of Cu to a rumen microbiota (Forsberg, 1978)
and of Ni to marine filamentous fungi (Babich and Stotzky, 1983e).
Salinity Increasing the salinity decreased the toxicity of Cd to a marine alga
(Wong et al., 1979) and of Ni to marine filamentous fungi (Babich and
Stotzky, 1983~).
Water hardness Increasing the hardness reduced the toxicity of Ni and Cd to filamentous
fungi (Babich and Stotzky, 198la, b), and of Zn to an alga (Wong, 1980)
and a paramecium (Chapman and Dunlop, 1981).
Chelating agents EDTA reduced the toxicity of Ni to an alga (Spencer and Nichols, 1983)
and an actinomycete (Babich and Stotzky, 1983e). of Cd to a cyano-
bacterium (Singh and Pandey, 1981), and of Cu to a bacterium (Love-
less and Painter, 1968) and algae (Sunda and Guillard, 1976).
Pyridinedicarboxylic acid reduced the toxicity of Ni to a bacterium, an
actinomycete, a yeast, and a filamentous fungus (Babich and Stotzky,
1983e).
NTA reduced the toxicity of Ni to algae (Spencer and Nichols, 1983), an
actinomycete, and filamentous fungi (Babich and Stotzky, 1983e) and
of Zn to a cyanobacterium (Greene ef al., 1978) and algae (Allen et a[.,
1980; Canterford and Canterford. 1980).
Succinic acid reduced the toxicity of Pb to filamentous fungi (Babich and
Stotzky, 1978a).
Citric acid reduced the toxicity of Ni to an actinomycete (Babich and
Stotzky. 1983e).
Undefined organics Proteose peptone reduced the toxicity of Cd to a protozoan (Houba and
Remacle, 1982).
TABLE I-Continued
Physicochemical
Yeast extract reduced the toxicity of Ni to a bacterium and a yeast (Ba-
bich and Stotzky. 1983e) and of Pb to tilamentous fungi (Babich and
Stotzky. 1979al.
Humic acids reduced the toxicity of Ni (Babich and Stotzky. 1982dl and
Pb (Babich and Stotzky, 1979a) to filamentous fungi and of Cd to an
alga (Gjessing, 1981).
Clay minerals Montmorillonite > attapulgite > kaolinite reduced the toxicity of Ni (Ba-
bich and Stotzky, 1982b. d, 1983b) and Pb (Babich and Stotzky. 1979a)
to filamentous fungi, with the extent of reduction being correlated with
the cation exchange capacity of the clays.
Montmorillonite and. to a lesser extent, kaolinite protected bacteria and
fungi against the toxicity of Cd (Babich and Stotzky. 1977b, c).
Hydrous metal oxides Fe(OH), reduced the toxicity of Cu to an alga (Steemann Nielsen and
Kamp-Nielsen, 1970).
A&O, . nHzO and MnOz * nH,O reduced the toxicity of Ni to a marine
filamentous fungus (Babich and Stotzky, 1983e).
Temperature Increasing the temperature increased the toxicity of Cd to a protozoan
(Szeto and Nyberg. 1979) and a chemolithotrophic bacterium (Kova-
lenko and Karavaiko, 1981) and of Hg to an alga (Huisman et al., 1980).
Increasing the temperature decreased the toxicity of Ni to a filamentous
fungus (Babich and Stotzky. 1982d).
Hydrostatic pressure Increasing the hydrostatic pressure increased the toxicity of Ni to an
unidentified marine bacterium capable of oxidizing Mn?’ (Arcuri and
Ehrlich, 1977).
mechanisms of action of the metals. Unlike the vast majority of organic mutagens
and carcinogens, metals do not require metabolic activation to be genotoxic. and
the ion (e.g., Cd’+) or the ion complex (e.g., CrOi-) appears to be the ultimate
form of the mutagen and carcinogen. Three important aspects of the mutagenic
or carcinogenic activity of heavy metals have not been fully clarified: (1) how the
metal ion or complex penetrates cell membranes; (2) how metals combine, once
inside the cell, with nucleic acids and, hence, alter the genetic information; and
(3) why a divalent heavy metal cation, such as Cd2+, is genotoxic, whereas other
divalent cations, such as calcium (Ca’+) or magnesium (Mg2+), are not genotoxic
(Furst, 1981). However, it has been suggested that because genotoxic metals (such
as Cd2+, Hg2+, and Pb2+) can form covalent bonds whereas essential cations
(such as Ca2+ and Mg2+) form ionic bonds, the genotoxic heavy metal ions form
more stable complexes with cellular components than do essential ions and, thus,
prevent normal cation-mediated functions (Christie and Costa, 1983).
Heavy metals have a strong affinity for cell surfaces. For example, Cd’+, Pb’+ ,
and Zn’+ sorbed strongly to the surfaces of amphibian kidney cells (Kiremidjian
and Stotzky, 1975), to macrophages and lymphocytes of guinea pig (Kiremidjian-
Schumacher et al., 1981a, b), to normal human lymphoid and Burkitt lymphoma
cells (Kiremidjian-Schumacher and Stotzky, 1976), and to bacteria and yeasts
(Collins and Stotzky, 1982). Although heavy metals differed in their affinities for
surface ionogenic groups of different cells, all reduced the surface charge of these
TABLE 2
INFLUENCE OF SOME PHYSICOCHEMKAL FACTORS ON THE TOXICITY OF HEAVY METALS TO
REPRESENTATIVES OF THE AQUATIC BIOTA
Physicochemical
PH Increasing the pH from 4.1 to 5.1 increased the toxicity of Cd and Cu to

the duckweed, Lemnu paucicostata (Nasu et al.. 1983).
Increasing the pH from 6 to approximately 8.4 reduced the toxicity of Cr6+
to the water flea, Duphnia magna (Muller. 1980).
Increasing the pH from 5.5 to 7.7 increased the toxicity ofV to the rainbow
trout, Salmo gairdneri (Stendahl and Sprague, 1982).
Inorganic cations Ca reduced the toxicity of Cd to the freshwater shrimp, Gammarus pulex
(Wright and Frain, 1981), and to the brook trout, Salvelinus fiontindis
(Carroll et nl., 1979).
Ca reduced the toxicity of Zn to the fathead minnow, Pimephales promelas
(Judy and Davies, 1979).
Zn reduced the toxicity of Cd to D. magna (Attar and Maly, 1982).
Inorganic anions Increasing the level of phosphate reduced the uptake and accumulation of
Pb by the freshwater amphipod, Hyallela azteca (Freedman et al., 1980).
Pyrophosphate reduced the uptake of Cd by D. magna (Poldoski, 1979).
Hardness Cu was 4 to 6 times more toxic to G. pulex in soft than in hard water
(Stephenson, 1983).
Cd was more toxic to eggs of the freshwater fish, Olyzias latipes, in soft
than in hard water (Michibata, 1981).
Increasing the total alkalinity or the hardness reduced the toxicity of Cd
(Chakoumakos et al., 1979) and Cr6+ (Muller, 1980) to D. magna.
Salinity Increasing the salinity from 16.5 to 33.0°/oo reduced the toxicity of Cu to
embryos of the Pacific oyster, Crnssostrea gigas (Coglianese, 1982).
Increasing the salinity from 1 to 35O/ooreduced the toxicity of Cd and Cr6+
to juveniles of the blue crab, Callinectes sapidus (Frank and Robertson.
1979).
Chelating agents EDTA reduced the absorption of Cu and Cd by L. paucicostata (Nasu et
al., 1983).
EDTA. citrate, oxalate, and glycine reduced the toxicity of Cu to embryos
of C. gigas (Knezovich et al., 1981).
EDTA, NTA. and diethylenetriaminepentaacetic acid reduced the uptake
and accumulation of Cd, Cu, Pb, and Zn by the carp, Cyprinus carpio
(Muramoto, 1980, 1981).
Uptake of Cd by the American oyster, Crussostrea virginica, was reduced
by EDTA and NTA (Hung, 1982).
Ethylenebis[N,N’-(2,6-dicarboxy)]piperidine, EDTA, and NTA decreased
the uptake of Cd by D. magna (Poldoski, 1979).
NTA and glycine reduced the uptake of Cu by larvae of the midge, Chi-
ronomus tentans (Dodge and Theis, 1979).
Undefined organics Humic acid reduced the uptake of Cu by C. tentans (Dodge and Theis,
1979), D. magna (Poldoski, 1979), and embryos of C. gigas (Knezovich
et al., 1981).
Particulates Uptake of Cd by the clam, Protothaca staminea, was reduced in the pres-
ence of sediment (Hardy et al., 1981).
Colloidal organic particulates decreased the uptake of Cd by the freshwater
crustacean, Simocephalus serrulatus (Giesy et al., 1977).
Temperature Increasing the temperature from 5 to 30°C reduced the toxicity of Cti+ to
the goldfish, Carassius auratus, and, to a lesser extent, to the golden
TABLE 2-Continued
Physicochemical
shiner. Notenzigorzus ctysoleucus, but did not affect the toxicity to the
bluegill, Leponzis macrochirus, the channel catfish, Ictuhrus puncfnf~~.
or S. g&&z& (Smith and Heath, 1979).
Fingerlings of the perch. Percafluviatilis, accumulated more Cd at I5 than
at 5°C (Edgren and Notter. 1980).
Increasing the temperature from 5 to 20°C increased the uptake and ac-
cumulation of Cd by C. virginica (Hung, 1982).
cells (Kiremidjian and Stotzky, 1975; Kiremidjian-Schumacher and Stotzky. 1976;

Kiremidjian-Schumacher et al., 1981a), affected immune reactions between an-
tigens and sensitized cells (Kiremidjian-Schumacher et al., 1981b), and reduced
the production of lymphokines and the ability of macrophages to destroy tumor
cells (Nelson et al., 1982).
Heavy metals can modify nucleic acids either by interacting directly with them
or affecting their synthesis. Heavy metal ions interact with nucleic acids strands
to influence their base pairing, as studies with synthetic polyribonucleotides have
shown that Cd2+ and Mn”, but not Zn2+, caused mispairing of nucleic acid
bases (Murray and Flessel, 1976). Using synthetic polyribonucleotide templates,
the fidelity of DNA replication by DNA polymerases from avian myeloblastosis
virus was decreased by Cd2+, Co2+, Cr6+, Cu+ , Cu?+ , Mn2+, Ni’+ , and Pb’+
(Sirover and Loeb, 1976, 1977) and that by DNA polymerase I from Escherichia
coli was decreased by Cd2+, Co2+, Hg’+, Mn*+, and Nil+ (Miyaki et al., 1977).
It has been stated that “in studying fidelity of DNA synthesis by DNA polymer-
ases, one is studying mutagenesis in vitro” (Zakour et al., 1981) Heavy metals
also inhibited RNA transcription, with the sequence of inhibition being Pb2+ >
Zn’+ > Cu2+ > Be2+ > Cd2+ > N?+ > Co?+ > Mn’+ (Niyogi et al., 1981).
Because of their interference with spindle formation, heavy metals have the
potential of inducing aneuploidy. Heavy metals interact strongly with the sulfhy-
dry1 groups involved in the polymerization of tubulin. thereby hindering the de-
velopment of microtubules and the formation of the spindle apparatus. Tin(Sn)
(i.e., trimethyltin chloride, tributyltin chloride), Hg (i.e., HgCI,, phenylmercuric
chloride, methylmercuric chloride, methylmercuric dicyandiamide), and Pb (i.e.,
Pb(NO,),, triethyllead chloride, trimethyllead chloride) interfered with spindle
formation and induced c-mitoses in the root tips of Alliurn cepa (Onfelt and
Ramel, 1979). Exposure of human lymphocytes to methylmercuric chloride, but
not to HgCl,, induced aneuploidy and caused c-mitoses and disturbed metaphases
(Kirsch-Volders et al., 1983).
In vitro exposures of mammalian cells to heavy metals have resulted in a variety
of chromosomal and chromatid aberrations, including gaps and breaks, and an
increased frequency of sister-chromatid exchanges (Table 4). In addition, the
transformation of cells in tissue culture by heavy metals has also been noted:
Cd2+ increased transformation in human diploid cells (strain L-58) infected with
TABLE 3
INFLUENCE OF SOME PHYSICOCHEMICAL FACTORS ON THE TOXICITY OF CADMIUM TO
LABORATORY ANIMALS”
Physicochemical
Zinc Zn protected against Cd-induced: (a) necrosis of mouse and rat testes (Par-
izek, 1957; Gunn et al.. 1961. 1968a, b); (b) formation of interstitial cell
tumors in mouse and rat testes (Gunn et al., 1963, 1964); (c) formation of
pleomorphic sarcomas in rats at the subcutaneous site of Cd injection
(Gunn et al., 1964); (d) placental necrosis and fetal death in mice (Chi-
quoine, 1965); (e) teratogenesis in hamsters (Ferm and Carpenter, 1967,
1968) and chicks (Narbaitz et al., 1983); (f) anemia in quail (Fox ef al.,
1971); (gf growth depression and gizzard abnormalities in chicks (Hill et
al., 1963); and (h) development of lesions on sensory ganglia of rats (Gab-
biani ef al., 1976).
Copper Cu protected against Cd-induced mortality in chicks (Hill ef al.. 1963) and
anemia in quail (Fox et al., 1971).
Lead Pb and Cd, both teratogens, interacted synergistically in their teratogenicity
to hamsters (Ferm, 1971).
Mercury Hg and Cd, both teratogens, interacted additively in their teratogenicity to
hamsters (Gale, 1973) but antagonistically in their teratogenicity to mice
(Layton and Ferm, 1980).
Iron Fe3+ protected against Cd-induced fetal growth retardation in mice (Web-
ster, 1979). mortality and growth depression in chicks (Hill et al., 1963),
and anemia in quail (Fox er al.. 1971).
Selenium Se protected against Cd-induced: (a) necrosis of mouse (Gunn et al.. 1968a,
b) and rat (Mason ef al., 1964) testes: (b) necrosis of the placenta of rats
(Parizek et al.. 1968): (c) teratogenicity in hamsters (Holmberg and Ferm,
1969); and (d) depression of blood hemoglobin and increase in heart size
of rats (Meyer et al., 1982).
Se blocked the Cd-induced inhibition of hepatic microsomal biotransfor-
mation of drugs, i.e., aniline and ethylmorphine (Early and Schnell, 1981).
Arsenic As and Cd, both teratogens, interacted additively in their teratogenicity to
hamsters (Holmberg and Ferm, 1969).
Cysteine Cysteine protected against Cd-induced anemia in quail (Fox et al., 1971) and
necrosis of mouse testes but potentiated the toxicity of lethal amounts of
Cd to mice (Gunn er al., 1966, 1968a. b).
Glutathione Glutathione protected against Cd-induced necrosis of mouse testes (Gunn
ef al., 1966) and lesions of sensory ganglia of rats (Gabbiani et al., 1976).
Ascorbic acid Ascorbic acid protected against Cd-induced growth retardation in chicks
(Hill, 1979) and anemia and growth retardation in quail (Fox ef al., 1971).
Chelating agents EDTA protected rats from Cd-induced hypertension (Yunice and Perry, 1961)
and lethal doses of Cd (Cantilena and Klassen, 1981).
2,3-Dimercaptopropanol protected against Cd-induced necrosis of mouse
testes (Gunn et al., 1968a) but had no effect on the mortality of mice
(Gunn et al., 1968a) or rats (Cantilena and Klassen, 1981) exposed to lethal
doses of Cd.
NTA protected pregnant rats from Cd toxicity but had little (Nolen et al.,
1972) or no (Scharpf et al., 1972) effect on reducing Cd teratogenicity.
D,L-Penicillamine, diethylenetriaminepentaacetic acid, and meso-dimercap-
tosuccinic acid reduced the toxicity to rats of lethal levels of Cd (Cantilena
and Klassen, 1981).
0 Cadmium was selected as the representative metal, as it is the most studied heavy metal, and
therefore there is more information about its toxicity as mediated by physicochemical factors than
for other metals.
TABLE 4
HEAVY METAL CLASTOGEN~CITYOBSERVED IN IN VITRO MAMMALIAN CELL ASSAYS
Cell line Comments and references
Human lymphocytes Chromosomal aberrations were not induced by Pb acetate (Schmid et al.,
1972).
Chromosomal aberrations were not induced by CdCl?. Co(NO,l,, and
HgClz (Paton and Allison, 1972).
Chromosome-breaking ability of several Cr-containing compounds fol-
lowed the sequence: K2Cr20, > K$rO, > Cr(CH,COO), > Cr(NO,),,
CrCl, (Nakamuro et al., 19781.
CdCl?, Pb acetate, and ZnClz did not affect the incidence of chromatid or
chromosomal gaps or of aneuploid cells; dicentric chromosomes oc-
curred only with ZnClz (deKnudt and Deminatti, 1978).
NiSO, increased the frequency of sister-chromatid exchanges and chro-
matid aberrations, including gaps and breaks (Larramendy et al., 1981).
Cd acetate, but not Pb or Zn acetate, induced chromatid aberrations, in-
cluding breaks. gaps, and acentric fragments (Gasiorek and Bauchinger,
1981).
K&r20,, but not CrCl,. induced a dose-dependent increase in the fre-
quency of sister-chromatid exchanges; K,Cr20, and, to a much lesser
extent. CrCl, induced chromosomal aberrations (Stella er a/., 1982).
K2Cr20, increased the incidence of chromosome and chromatid aberra-
tions and the incidence of sister-chromatid exchanges (Imreh and Rad-
ulescu. 1982).
Methyl Hg, but not HgC&,. increased the incidence of c-mitoses, disturbed
metaphases, and chromosomal aberrations and induced aneuploidy
(Kirsch-Volders ei ul., 1983).
Syrian hamster K2Crz0, and CrO,. but not CrC&, CrCI,, or Cr,(SO,),, induced chromatid
embryo cells gaps. breaks, and exchanges (Tsuda and Kato, 1977).
NiSO, increased the incidence of sister-chromatid exchanges and chro-
matid aberrations, including gaps and breaks (Larramendy et al.. 1981).
Chinese hamster K,Cr,O, and NazCrzO, induced chromosomal aberrations, including chro-
ovary cells matid gaps, breaks, and interchanges, and increased the frequency of
sister-chromatid exchanges (Majone and Levis, 1979).
Chinese hamster CdSO, induced c-mitoses and chromosomal aberrations, particularly gaps
tibroblast cells (Rohr and Bauchinger. 1976).
K,Cr20,, but not CrCl, or Cr(NO&, increased the incidence of sister-
chromatid exchanges-(Venier et cd., 1982).
Chinese hamster K2Cr10, induced chromatid breaks and exchanges and isochromatid dele-
lung cells tions (Newbold et al.. 1979).
Cu(NO,l, increased the frequency of sister-chromatid exchanges (Sideris
and Petraki, 1982).
K,CrO,, Na&rO,, Na2.Cr20,. and, to a much lesser extent, CrCI, and
Cr,O, increased the Incidence of sister-chromatid exchanges (Elias et
al., 1983).
Don Chinese CrCl,, CrO,, K,Cr,O,. K,CrO,, NiSO,, and NiC&, but not Cr,(SO,),.
hamster cells FeSO,, FeCl,, Cd(NO,),, CdCl?. Cd acetate, SnC&. HgCl?. Hglz, or Hg
acetate, increased the frequency of sister-chromatid exchanges (Ohno
et ul., 1982).
FM3A cells from a K,CrO,, K2Cr207. and CrO, induced large numbers of chromosomal aber-
mouse mammary rations, mcluding breaks and exchanges, whereas Cr2(S0,), induced
carcinoma only a few chromosomal aberrations; KMnO, and, to a lesser extent,
MnClz induced chromosomal aberrations: K,Ni(CN), and, to a lesser
extent, NiS induced chromosomal aberrations, whereas NiClz and Ni
acetate induced little increase in chromosomal aberrations; no chro-
mosomal aberrations occurred with CdCl? or HgCl: (Umeda and Nish-
imura. 1979).
the oncogenic mammalian leucosis virus (Zasukhina et al., 1977), and Cd*+, Cr6+,
and, to a lesser extent, Co*+, Cu*+, Hg2+, Mn*+, Ni*+, Pb*+, and Zn*+ enhanced
the transformation of Syrian hamster embryo cells after the inoculation of the
simian adenovirus, SA7 (Cast0 et al., 1976, 1979).
IN V/T/30 ASSAYS OF THE MUTAGENICITY OF HEAVY METALS
Several mammalian systems have been developed as in vitro assays of the
mutagenicity of heavy metals. Mutagenicity is most commonly detected by al-
terations in the expression of genes specifying enzymes that control the incor-
poration of an added toxic base analog: a positive mutagenic effect is indicated
by the loss of the marker enzyme, enabling cultured tester cells to grow and form
colonies in the presence of the toxic antimetabolite. Commonly used enzyme
markers include hypoxanthine-guanine phosphoribosyl transferase (HGPRT), the
loss of which imparts resistance to 6-thioguanine or 8-azaguanine, and thymidine
kinase (TK), which incorporates such thymidine analogs as 5bromodeoxyuridine
or trifluorothymidine (Heck and Costa, 1982). Cd*+ and Ni*+ induced trifluoro-
thymidine-resistant mutant cells in an assay using L5178Y/TK mouse lymphoma
cells (Amacher and Paillet, 1980). Cr6+, but not Cr3+, Cd*+, Hg2+, Mn*+, or
Ni*+, induced 8-azaguanine-resistant mutant cells in FM3A cells derived from a
mouse mammary carcinoma (Nishimura and Umeda, 1978) and in Chinese ham-
ster cells as soluble K2Cr,07 or ZnCrO, but not as insoluble PbCrO, (Newbold
ef al., 1979). Mn*+ was highly mutagenic in the induction of g-azaguanine-resis-
tant mutants in Chinese hamster cells, whereas Ni*+ and Co*+ were only weakly
mutagenic (Miyaki et al., 1979). Cd*+, Cr6+, Mn*+, and, to a lesser extent, Hg*+
and Pb*+ induced forward mutation at the TK locus in L5 178Y mouse lymphoma
cells (Oberly et al., 1982).
Perhaps the most information about the mutagenicity of heavy metals has been
obtained with microbial assays (Table 5), with the majority of studies using either
the Salmonella typhimurium or the Escherichia coli reverse mutation system.
The S. typhimurium assay utilizes mutant strains auxotrophic for histidine (his-)
that have either a normal or an error-prone DNA repair capability, and therefore,
both base-pair substitutions and frameshifts can be detected. Exposure to mu-
tagenic agents induces reversion to histidine prototrophy, and clonies of such
revertants develop on histidine-free media. The addition of a microsomal fraction
is needed to activate many organic mutagens (Ames, 1971). The E. coli reverse-
mutation system uses a strain (usually, strain WP2) that requires tryptophan for
growth and reverts to tryptophan nondependence after exposure to mutagens
(Green and Muriel, 1977). Another system, the Bacillus subtilis ret-assay, has
been used to study the ability of heavy metals to damage DNA by measuring the
inhibition of growth of DNA repair-deficient bacteria (Nishioka, 1975).
Although mutagenicity assays, in particular the Salmonella reverse-mutation
assay, are routinely used to predict the potential carcinogenicity of chemicals,
the results obtained with heavy metals are inconsistent. For example, Cd (De-
graeve, 1981), Cr (Leonard and Lauwerys, 1980), Ni (Leonard et al., 1981), and
Pb (Gerber et al., 1980) are carcinogens, yet only Cr is consistently detected as
being mutagenic, and then only as Cr 6+ (Table 5). Several reasons have been
suggested for the inconsistency of microbial mutagenicity assays for detecting
heavy metal genotoxicity. For example, it was suggested that (1) the bacterial
strains in current use are able to correct errors in DNA synthesis by mismatch
repair and, thus, are genetically incapable of yielding a positive mutagenic re-
sponse to heavy metals: (2) carcinogenic heavy metals may be comutagens rather
than mutagens; (3) the bacteriocidal effects of heavy metals may obscure their
mutagenicity, especially if the results of these experiments are expressed as the
number of mutants/plate without considering survival levels; and (4) there may
be no correlation between the carcinogenicity and mutagenicity of heavy metals
(Rossman, 1981). Nevertheless, the reverse-mutation assay with Corynebacte-
rium sp. was sensitive to NI ‘2+ (Pikalek and Necasek, 1983) and that with S.
typhimurizcm (Petrilli and deFlora, 1977) and E. coli (Green et al., 1976) was
sensitive to Cr6+, and the B. subtilis ret-assay was sensitive to Cd?+, Co?+, Cr6+,
Hg+, Hg*+. and organic Hg (Kanematsu et al., 1980).
Another explanation for the inconsistency of microbial mutagenicity assays, as
well as of mammalian cell cultures assays for clastogenicity, is that the compo-
nents of the assay media detoxify the heavy metals (McCann et al., 1975). The
assay media contain (1) phosphates which can precipitate heavy metals, (2) or-
ganic chelators which may reduce the bioavailability of heavy metals. and (3)
essential cations, such as Ca*+ and Mg?+, which can compete with the added
heavy metals for sites on cell surfaces. This explanation seems most plausible,
as Cr6+, the only metal consistently detected as being mutagenic, occurs as an
anionic complex (either as CrO$- or Cr,O:-) whose bioavailability is not as
affected by phosphate, organic chelators, or cations in the assay media. Similarly,
studies with several heavy metals using FM3A ceils from a mouse mammary
carcinoma showed that the greatest clastogenic effects occurred with anionic
complexes of heavy metals (i.e., Cr#, CrOi-, Ni(CN)i-, MnOi-) rather than
with their cationic forms (i.e., Cr3+, Ni’ +, Mn2+) (Umeda and Nishimura, 1979).
Furthermore, heavy metals exhibited differential affinities for the organics com-
monly incorporated into microbiological and other assay media: the sequence for
the affinity of Hg was casamino acids > proteose peptone > yeast extract >
Bacto tryptone > Bacto peptone; for Cu it was casamino acids > yeast extract
> Bacto tryptone > proteose peptone > Bacto peptone; for Pb it was casamino
acids > yeast extract > Bacto tryptone > Bacto peptone > proteose peptone;
and for Cd it was casamino acids > proteose peptone > Bacto tryptone > yeast
extract, with no binding to Bacto peptone (Ramamoorthy and Kushner, 1975).
As the extent of binding of heavy metals to organics affects the toxicity of the
metals (see Tables 1 to 3), the differential affinities of the heavy metals for specific
organics and the different kinds of organics that are incorporated into media of
different assay systems may also explain the inconsistent results obtained in mu-
tagenicity and clastogenicity assays of heavy metals.
The influence of the composition of the assay media on the toxicity and mu-
tagenicity of heavy metals is best illustrated with Cr, as Cr3+ and Cr6+ exhibit
differential toxicities (Babich et al., 1982a, b; Venier et al., 1983) and mutage-
nicities (deFlora, 1978; Petrilli and deFlora, 1978a, b; Venier et al., 1982) in
microorganisms and in mammalian cells. Cr3+ is the most stable oxidation state
TABLE 5
MUTAGENICITY OF HEAVY METALS AS DETECTED IN MICROBIAL SYSTEMS E
Biologic system Results References

E. coli B/r/Sd-4 Mn2+ and Cu?+, but not Hg?+, induced mutation from streptomycin Demerec and Hanson, 1951;
dependence to streptomycin independence Demerec et al., 1951
B. subtilis Trp Cu2+ was mutagenic; inhibited transformability Weed, 1963
transformation assay
E. co/i K-12 (A) induct test Cu’+ was ineffective as an inducer of phage production by lysogenic Heinemann and Howard. 1964
E. coli
Coliphage T4 Mn?+ induced mutation at the rlI locus Orgel and Orgel, 1965
E. coli K-39 (A) induct test Methyl Hg+ and methoxyethyl Hg+ were ineffective inducers of Fiskesjo. 1971
phage production by lysogenic E. coli i;
S. cerevisiae Mn’+ induced antibiotic-resistant mutation in mitochondrial DNA Putrament et al., 1973, 1975a, b, 1977 *z
E. coli WP2 Tip reversion Cr6+ was mutagenic; Cr3+, Cd’+, Hg2+, and Zn2+ were not Venitt and Levy, 1974
E
assay
Platymonas subcordijormis Pb?+ was not mutagenic Hessler, 1975 5
morphotype assay 5
B. subtilis ret-assay Cr6+, organic Hg, Cd*+, and Mn2+ were mutagenic; Cr3+, Co2+ Nishioka, 1975 ”v)
Cu+, Cu2+, Fez+, Fe3+, Ni2+, Pb2+, and Zn2+ were not b
B. subtilis ret-assay; S. Organic Hg- and Ni- containing fungicides were not mutagenic Shirasu et al., 1976
typhimurium His- and E.
coli Tip reversion
assays
Schizosaccharomyces pombe Mitotic ade- gene conversion induced by CI-6+ Bonatti et al., 1976
S. typhimurium His- Fe2+ induced mutation; mutagenicity enhanced by the addition of Brusick et al., 1976
reversion assay microsomal fraction
E. coli WP2 Trp- reversion CI-6+, but not Ni2+, was mutagenic Green et al., 1976
fluctuation assay
S. typhimurium His- Zn*+ , but not Cd’+, was highly mutagenic in the presence of a Kalinina and Polukhina, 1977
reversion assay mouse liver microsomal fraction
S. typhimurium His- C1-6+ was mutagenic, but mutagenicity was not enhanced by addition Petrilli and deFlora, 1977
reversion assay of microsomal fraction; Cr3+ was nonmutagenic, either with or
without microsomal fraction
E. coli WP2 alkaline Cr6+ degraded bacterial DNA in vivo Nishioka and Yagi, 1978
sucrose gradient
sedimentation assay
S. typhimurium His- Cr6+, but not Cr3+. was mutagenic Lofroth and Ames. 1978
reversion assay
S. typhimurium His- and Cr6+ was mutagenic Tindall et al., 1978
E. co/i WP2 Tip
reversion assays
E. coli HS30R Arg- Cr6+ was highly, but Cr3+ was slightly, mutagenic Nakamuro et al., 1978
reversion assay; B.
subtilis ret-assay
Plectonema boryanrtm Mn?+ induced phage-resistant, but not streptomycin-resistant, Singh and Kashyap, 1978 3
mutants
2
S. typhimurium His Addition of a complete microsomal fraction decreased the Lofroth. 1978; Gruber and Jennette.
reversion assay mutagenicity of CI-6’ ; mutagenicity was not affected by addition of 1978 ?
a microsomal fraction lacking NADPH 8
S. typhimurium His- Numerous @+-containing compounds were mutagenic, with deFlora, 1978: Petrilli and deFlora,
reversion assay mutagenicity decreasing upon addition of a complete microsomal 1978a; deFlora et al., 1982
fraction, reducing agents (ascorbic acid. sodium sulfite), or reduced
metabolites (DPNH, TPNH, reduced glutathione); addition of an
oxidizing agent (KMnO,) prevented reversal of mutagenicity by the
complete microsomal fraction
S. typhimrcrium His Numerous Cr3+-containing compounds were nonmutagenic, in the Petrilli and deFlora, 1978b
reversion assay absence or presence of a mixed microsomal fraction; addition of an
oxidizing agent (KMnO,) induced mutagenicity
E. coli Pol A- survival test: PbCrO, was not mutagenic Nestmann et (I/.. 1979
E. coli WP2 Trp-
reversion assay; E. coli
K-12 Gal+ forward
mutation assay
S. typhimurium His- PbCrO, was mutagenic, with mutagenicity being due to the Crh+, and Nestmann et rrl.. 1979
reversion assay; E. coli not to the Pb’+, component
WP2 reversion fluctuation
assay; S. cerevisiae D5
mitotic recombination assay
S. typhimurium His- Pb” was not mutagenic Simmon, 1979: Rosenkranz and
reversion assay; Poirer. 1979
E. coli Pol A- assay
B. subtilis ret-assay Mutagenicity of Cr6’ was not enhanced by Hg?’ Matsui, 1980
S. typhimurium His- Gastric juice decreased the mutagenicity of Cr6+ deFlora and Boido, 1980
reversion assay
TABLE 5-Continued K
Biologic system Results References
S. cerevisiae D4 Cd”’ induced ade- and trp- gene conversions Eger and Doko, 1980
B. subtilis ret-assay Cd?+, Co*+, Cr6’, Hg+, Hg?+, and organic Hg were mutagenic; Kanematsu et ul., 1980
Cu+, Cu*+, Cr3+, Ni?+, Pb’+, and Zn?+ were not
E. coli WP2 Trp Her- Cr6+ was mutagenic Kanematsu et al., 1980
reversion assay
S. typhimurium His- cis-[Cr(bipyridine)?oxylate]I . 4H?O, cis-[Cr(bipyridine)zCIJCI . Warren et ~1.. 1981
reversion assay; E. co/i 2Hz0, [Cr(urea)6]C13 . 3Hz0, and cis-[Cr(l,lO-
K-12 differential repair phenanthroline)&]Cl . 2.5H20 were mutagenic in both assays;
assay nine other Cr3+-chelates were nonmutagenic in either assay: and
six others were mutagenic in only one of the assays
S. typhimurium His- spot Cr6+ was mutagenic; Co*+, Cu2+. Cd?+, Cr3+, Mn?+, and Ni?+ were Tso and Fung, 1981
test reversion assay not
B. subtilis and S. The chelators, EDTA, salicylate. and Tiron, reduced the mutagenicity Gentile et al., 1981
typhimurium ret-assays of CP+ ; citrate and salicylate increased the mutagenicity of Cr3 +
S. typhimurium His- trans-[Co(4-picoline),C121Cl, trans-[Co(3-picoline),C12]Cl, cis-[Co( I, IO- Schultz et al., 1982
reversion assay; E. coli phenanthroline)2C121Cl, and cis-[Co(l.IO-phenanthroline)zoxylate]Cl
K-12 differential repair were mutagenic; nine other Co r + -chelates were not mutagenic in
assay both assays
E. coli Pol A- assay Cd2+ and Cuz+ were mutagenic Babich and Stotzky, 1982e
S. cerevisiae D7 Mn2+ and Cr6+ induced mitotic gene conversion at the trp- locus Singh, 1983
and reverse mutation at the ilv- locus
S. typhimurium His- Cr6+, but not Cr3+, was mutagenic Venier et al., 1983
reversion assay
Corynebacterium sp. 887 N?+ was mutagenic; Cd’+ and Co?+ were not Pikalek and Necasek, 1983
Horn- reversion assay
S. typhimurium His- Cu2+ alone or ascorbic acid alone was not mutagenic, but a Norkus et al., 1983
reversion assay combination of Cu?+ and ascorbic acid was mutagenic, and its
mutagenicity could be blocked by EDTA
S. typhimurium His- Cr3+, as chromatin B and chromatin MS-both tannin agents-was Bronzetti et ul., 1983
reversion assay mutagenic only after metabolic activation
S. typhimurium His- 12 @+-containing compounds were, but one Cr”- and three Cr3+- Petrilli and deFlora, 1983
reversion assay containing compounds were not, mutagenic
S. typhimurium His- Crb+ mutagenicity was decreased by rat liver microsomal fraction Bennicelli e/ al., 1983
reversion assay and, to a lesser extent, by human lung microsomal fraction
of Cr, and it has a tendency to form complexes with organic ligands, such as
proteins and nucleic acids. Because of this strong binding tendency, Cr3+ does
not easily penetrate cells and is frequently bound to extracellular constituents
(Gentile et af., 1981). For example, when entering the blood stream, Cr3+ binds
strongly to plasma proteins, whereas Cr6+ concentrates inside erythrocytes (Gray
and Sterling, 19.50). Cr6+, as an anionic complex, crosses cell membranes by
passive or facilitated transport, utilizing the phosphate (PO:-)- or sulfate (SO:-)-
transport system. After Cr6+ enters the cell, it may be reduced to Cr3+, which
can bind to DNA and exert a genotoxic effect (National Academy of Science,
1974; Tsapakos and Wetterhahn, 1983). When Crh+ was preincubated with mi-
crosomal fractions from rat (deFlora, 1978; Gruber and Jennette, 1978) or rainbow
trout (deFlora et al., 1982) liver or with human erythrocyte lysates (Petrilli and
deFlora, 1978a), its mutagenicity in the Saltmxzefla reversion assay was elimi-
nated, apparently as a result of the reduction of Crhf to Cr3+, as incubation of
Cr6+ with a microsomal fraction from rat liver lacking NADPH did not eliminate
the mutagenicity of Cr 6+ (Lofroth, 1978). Furthermore, the addition of reducing
agents (ascorbic acid, sodium sulfite) or reduced metabolites (reduced gluta-
thione, DPNH, or TPNH) prevented mutagenesis by Cr6+, whereas the addition
of an oxidizing agent (KMnO,) inhibited the reversal of mutagenicity by micro-
somal fractions from rat liver and by human erythrocyte lysates (Petrilli and
deFlora, 1978a). Cr3 + was not mutagenic, and additions of microsomal fractions
from rat liver, lung, or muscle, rat muscle mitochondria (with or without ATP),
oxidized glutathione, or human serum, plasma, or erythrocyte lysates did not
induce mutagenicity. However, the addition of KMnO,, which oxidized Cr”+ to
Cr’+, induced mutagenicity (Petrilli and deFlora, 1978b).
EFFECT OF PHYSICOCHEMICAL FACTORS ON THE MUTAGENICITY AND

CLASTOGENICITY OF HEAVY METALS
There is essentially no information about the influence of abiotic factors on
heavy metal mutagenicity and clastogenicity, although studies of the toxicity of
various heavy metals, using microbes (Table I), representative aquatic plants and
animals (Table 2), or laboratory animals (Table 3). have clearly demonstrated the
mediating influence of such physicochemical factors. The rigid standardization of
mutagenicity and clastogenicity assays, which presumably is needed to ensure
repeatability of experiments and duplicability of data between laboratories, has
apparently hindered the elucidation of the potential mediating influence of abiotic
factors on heavy metal genotoxicity. For example, the finding that pH influenced
the mutagenicity of diethylnitrosamine and some polycyclic hydrocarbons and
aromatic amines prompted the investigators to state: “Since strong mutagenicity
is not always obtained at pH 7.4 (the value at which most microbial assays are
normally performed), pH may be an important variable to consider in determining
the mutagenic potential of suspected carcinogens. The range of mutagens tested
in which pH effects have been observed suggests that this variable may be of
general significance in microbial assays” (Leonardo and Peak, 1982).
Little is known about the influence of pH on metal-induced mutagenesis. The
mutagenicity of Mn2+ towards extranuclear (i.e., mitochondrial) DNA in Sac-
charomyces cerevisiae was pH dependent, as increasing the pH from 6 to 6.7

reduced the number of cytoplasmic petite (rho-) mutants and from 6 to 6.5 re-
duced the number of erythromycin-resistant mutants (Putrament et al., 1975a).
However, pH did not influence the mutagenicity of Cr6+ to S. typhimurium, as
no differences in the extent of revertants to histidine prototrophy were noted
after preincubation of CI-6+ at pH 7.4 or at pH 1.3 (deFlora and Boido, 1980).
Although little is known about the mediating influence of pH on heavy metal
mutagenicity, several studies have shown that pH influences the toxicity of heavy
metals to microbes (Table 1). However, the mechanisms whereby pH affects
heavy metal toxicity have not been clearly defined, as pH affects several aspects
of the interactions between cells and heavy metals. (1) pH affects the metabolic
state of the target cells, and differences in a specific biotic response to a heavy
metal at different pH values may reflect the altered physiology of the cells at
different pH values. (2) pH affects the adsorption and subsequent uptake of heavy
metals by cells, with metal uptake usually increasing as the pH is increased. For
example, increasing the pH of the medium from 3 to 7 increased the uptake of
Cd by living and heat-killed cells of Chlorella regularis (Sakaguchi et al., 1979)
and from 4.5 to 6.8 increased the uptake of Cd, Co, Cu, Mn, and Ni by Klebsiella
pneumoniae (Rudd et al., 1983). (3) pH affects the chemical speciation of some
heavy metals, e.g., at pH 7, Ni occurs as Nr ‘2+, Cd as Cd2+, Pb as PbOH+, Zn
as a mixture of Zn2+ and Zn(OH),, and Hg as Hg(OH),, and different inorganic
speciation forms of the same heavy metal have different toxicities (Babich and
Stotzky, 1977a, 1980a). (4) pH affects the extent of binding of heavy metals with
inorganic and organic ligands, and free and complexed heavy metals have dif-
ferent toxicities (Babich and Stotzky, 1983a).
The standardization of microbial assays for mutagenicity has limited the appli-
cability of these assays to evaluating the potential carcinogenicity and terato-
genicity of heavy metals in natural environments in which pH varies greatly. The
pH of most soils ranges between 5 and 7, but acid mineral soils may have a pH
of 4 and peat soils may be as low as pH 3, whereas the pH of calcareous soils
may be 8 and above (Lyon and Buckman, 1943). The pH range of lakes that have
some degree of flow through the basin is generally from 6 to 9. However, in
limestone regions, the dissolved carbonates (CO:-) and the Mg2+ and Ca2+ ions
may raise the pH to above 9, whereas in lakes in regions of lowlands and bogs,
the pH ranges from 4 to 6. The pH of seawater at the surface is very stable,
usually ranging from 8.1 to 8.3 (Reid, 1961).
The human body is composed of several microenvironments, each with its own
unique physicochemical properties, including pH. The pH of gastric juice varies
from 1.0 to 3.5, of urine from 4.5 to 7.8, of saliva from 6.0 to 7.0, of thyme in
the small intestine from 7.0 to 8.0, of blood from 7.2 to 7.4, of secretions from
the large intestine from 7.5 to 8.0, and the pH of bile is 8.0 (Guyton, 1971; Strand,
1983). Preincubation of Cr6+ in human gastric juice decreased its mutagenicity
as assayed with S. typhimurium: preincubation of Cr6+ in gastric juice at pH 1.3,
in heated gastric juice at pH 1.3, and in gastric juice adjusted to pH 7.4 resulted
in 318 + 28, 342 + 31, and 454 2 34 revertants to histidine prototrophy, re-
spectively, whereas preincubation in phosphate-buffered saline at pH 7.4 and in
sodium chloride adjusted to pH 1.3 resulted in 890 ? 42 and 823 * 37 revertants,

respectively. There is apparently a thermoresistant factor in human gastric juice
that inactivates Cr6+, possibly by reducing it to Cr3+, and whose inactivating
ability is greater at pH 1.3 than at pH 7.4 (deFlora and Boido, 1980). Chromates
have been associated primarily with respiratory cancer in human beings (Enter-
line, 1974), and the inactivation of chromates by human gastric juice may explain
the lack of an association between chromates and stomach cancer (deFlora and
Boido, 1980). Another beneficial result of the reduction of Cr6+ to Cr3+ is that
Cr3+ is only poorly absorbed from the intestines (Donaldson and Barreras, 1%6).
The mutagenicity of heavy metals is influenced by complexing organics, as is
their toxicity (Babich and Stotzky, 1980a, 1983a, b; Jakobsen and Eik-nes, 1982;
Stotzky and Babich, 1980, 1983, 1984; Sugawara er al., 1983). The mutagenicity
of Cr6+ was decreased, as assayed with the B. subtilis ret-assay, by increasing
concentrations of the chelating agents, ethylenediaminetetraacetic acid (EDTA),
salicylate, or Tiron (disodium-1,2-dihydroxybenzene-3,5disulfonate). However,
as Cr6+ was assayed as the anionic complex, Cr,O:-, and as chelating agents
interact only with cations, the protective effect of these chelators on the muta-
genicity of Cr,O:- could not have resulted from a direct interaction between the
chelator and the anionic complex of Cr. The chelators apparently reduced Cr6+
to Cr’+, which, as a cation, was then chelated. However, salicyiate and citrate,
both acting as chelators, induced a small mutagenic response with Cr3+, which
usually is not mutagenic, in the B. subtilis ret-assay. Nitrilotriacetic acid (NTA),
EDTA, Tiron, pyrophosphate, and acetate did not induce the mutagenicity of
Cr3+ (Gentile et al., 1981).
Seventeen Cr3 + -containing compounds were tested for their DNA-damaging
capabilities using an E. co/i differential repair assay and for their mutagenic-
ity using the S. typhimurium reversion assay. CrCI,, K3[Cr(oxylate),],
[Cr(NH3),H,0Cl]C1z, [Cr(NH3),oxylatelN03. [Cr(NH3)4Cl,lCl, [Cr(NH3)5CllC12,
K,[Cr(CN),], cis-[Cr(ethylenediamine)+&]Cl, and [Cr(NH3),H,0)]C13 were neg-
ative in both assays; cis-[Cr( 1, 10-phenanthroline)zCIZ]C1, cis-[Cr(bipyridine), oxy-
late]I, cis-[Cr(bipyridine)2ClZ]C1, and [Cr(urea),]Cl, were active in both assays;
and [Cr(ethylenediamine)31(SCN),, [Cr(ethylenediamine),]C13, [Cr( 1,2-propanedia-
mine),]Cl,, and trans-[Cr(ethylenediamine)&SCN),]SCN were less active in the
differential repair assay and negative in the mutation assay (Warren et al., 1981).
In a subsequent study, fifteen Co3 + -containing compounds were testing for their
mutagenicity and DNA-damaging properties. truns-[Co(4-picoline),C12]Cl, cis-
[Cot 1, IO-phenanthroline),oxylate]Cl, and cis-[Co( 1, IO-phenanthroline),C1,]Cl
were active in both assays; (+)-[Co(ethylenediamine)$1,, (+)-[Co(eth-
ylenediamine)3]13, ( - )-[Co(ethylenediamine),ll,, [cO(NH3),]c13, and [Co( 1 ,%-pro-
panediamine),]Cl, were active only in the DNA-repair assay; [Co(ethylene-
diamine),oxylate]Cl, (-) - [Co(ethylenediamine],oxylate]Cl, cis - [Co(ethylenedia-
mine),Cl,]Cl, truns-[Co(ethylenediamine),Clz]Cl, [runs-[Co(pyridine),CIZ]Cl,
and cis-[Co(2,2’-bipyridine)&lZ]C1 were only slightly active in the DNA-repair
assay and were not active as mutagens. truns-[Co(3-picoline),C12]Cl was the only
complex that was active in the Salmonella mutagenicity assay, but it was inactive
in the E. coli differential repair assay (Schultz et al., 1982).
The genotoxicity of heavy metals is apparently not necessarily decreased by

their interaction with organics. Heavy metals probably exist in biological systems
predominantly as metal-organic complexes, as a result of their interactions with
numerous organic molecules that possess moieties capable of chelation (e.g.,
carboxyl and amino groups) and binding (e.g., sulfhydryl groups). Chelation may
facilitate the transport of heavy metals to or from vulnerable target sites or hinder
those intracellular interactions that ultimately lead to cancer. The “carcinogenic
heavy metals” have the ability-via chelation or mixed (ternary) complex for-
mation-to interact with DNA and other nuclear constituents and to alter cellular
properties, such as membrane integrity. Some important factors controlling the
formation and activity of metal chelates in biological systems include: (1) intrinsic
stability of the chelate-metal complex, (2) chelate-metal ratios, (3) stability of
the chelate-metal complex as a function of pH, (4) competition from other metals
and ligands, both endogenous and exogenous, (5) overall charge and lipophilicity,
and (6) rates of chelate formation and hydrolysis of heavy metals (e.g., Cr un-
dergoes a series of reactions with water to form complex anionic species, such
as Cr,O:-). Furthermore, heavy metal ions in biological systems usually combine
simultaneously with two different ligands (i.e., ternary complexes). For example,
Cu2+ is bound simultaneously to two different amino acids in serum, with the
most usual mixed complex being Cu-histidine-cysteine (Martell, 1981; Schubert,
1981). The chemical properties of such ternary speciation complexes may have
a marked effect on the ability of a heavy metal ion to interact with target mole-
cules and cause a genotoxic or carcinogenic effect.
In natural environments, heavy metals are also complexed with organics, both
soluble (e.g., chelators) and particulate. For example, the humic acids present in
soils (Stevenson, 1972) and those occurring in freshwater (Wilson, 1978) and
marine (Rashid, 1971) sediments and in the aqueous phase of aquatic environ-
ments are capable of complexing variable amounts of metals. Chelated heavy
metals are usually less toxic than the free forms of the same heavy metals (see
Table 1 to 3; Cantilena and Klassen, 1981, 1982a, b; Babich and Stotzky, 1983e;
Planas-Bohne and Lehmann, 1983). However, no research has yet evaluated the
influence of organics occurring in natural environments on the genotoxicity of
heavy metals. Although the studies of Gentile et al. (1981), Warren ef al. (1981),
and Schultz et al. (1982) mentioned above indicate that, in general, chelation
reduces the mutagenicity of heavy metals, some exceptions were noted (e.g.,
citrate activated the mutagenicity of Cr3+). The organics present in tissue culture
and microbial assay media bind heavy metals (Ramamoorthy and Kushner, 1975),
thereby also reducing the reliability of standardized microbial mutagenicity and
tissue culture clastogenicity assays for detecting the mutagenesis and clasto-
genesis of heavy metals (McCann et al., 1975).
Inorganic cations and anions mediate the toxicity of heavy metals (Tables 1 to
3), and thus, they have the potential of affecting the genotoxicity of heavy metals.
It has been suggested (McCann er al., 1975) that the inorganic composition of
microbiological and tissue culture assay media is another factor that reduces the
reliability of standardized microbial mutagenicity and tissue culture clastogenicity
assays for detecting the genotoxicity of heavy metals. The cations normally
present in the assay media (e.g., Ca*+ and Mg*+) may reduce the genotoxicity
of heavy metals, as competition for sites on cell surfaces between essential cations
and the cationic speciation form(s) of heavy metals affects the extent of uptake
of the heavy metals by the cells. For example, the uptake of Cd*+ and Ni*+ by
E. co/i was reduced by Mg*+ (Abelson and Aldous, 1950). Essential inorganic
anions in the media also mediate the bioavailability of heavy metals. PO:- and
CO:- interact with the cationic form of heavy metals to form insoluble precipi-
tates, which are unavailable for uptake by the biota. For example, the cytotoxicity
of Pb to fungi was reduced in the presence of PO:- and CO:-, presumably due
to the formation of insoluble Pb3(P0,), and PbCO, (Babich and Stotzky, 1979a).
Furthermore, essential inorganic anions in the media may also compete with those
heavy metals that occur as anionic complex ions for sites on cell surfaces: e.g.,
SOi- reduced the uptake of CrOz- by human leukocytes (Jennette, 1981).
The effects of essential inorganic cations and anions on the genotoxicity of
heavy metals are not unique to microbiological and tissue culture assay media,
but they are applicable also to natural environments and to the various fluids and
tissues of the human body. The soil solution is essentially a weak electrolyte
composed of a variety of inorganic cations and anions of different valences
(Stotzky, 1972). Studies with terrestrial plants have noted that increasing the
concentration of potassium (K+), Ca*+, or Zn*+ decreased the uptake of Cd*+
(see Babich and Stotzky, 1978a). In fresh waters, CO:- and SOi- are the dom-
inant anions and Mg*+ and Ca* + the dominant cations, comprising 48 and 15%,
respectively, of the total cations in soft waters and 53 and 34%, respectively, in
hard waters (Reid, 1961). The toxicity of heavy metals is dependent on the hard-
ness of fresh water, with toxicity decreasing as hardness is increased (Table 1 and
2). The relation between water hardness and the genotoxicity of heavy metals in
aquatic environments has not been studied.
Seawater is a strong electrolyte solution (essentially a 3.5% salt solution), con-
sisting of 19.39 g/kg chloride (Cl-), 10.77 g/kg sodium (Na+). 2.71 g/kg SO:-,
1.30 g/kg Mg2+, 0.409 g/kg Ca*+, 0.388 g/kg Kf. and 0.23 g/kg CO:- (Tait and
desanto, 1972). The ameliorating effects of seawater on Ni toxicity to marine
fungi was related to the high concentration of Mg*+, which apparently competi-
tively inhibited uptake of Ni*+ by the fungi (Babich and Stotzky, 1983e). Cll,
the dominant inorganic ion in seawater, affects the speciation of some heavy
metals: e.g., in seawater, Hg occurs as a mixture of HgCl 7 and HgCli- and Cd
as a mixture of CdCl+, CdCl,, and CdCl; (Hahne and Kroontje, 1973). Although
there appear to be no studies on the comparative mutagenicities of Hg and Cd in
aquatic environments having different chlorinity (i.e., salinity), toxicity studies
have shown that Hg and Cd are less toxic in marine than in fresh waters, with
the differences in toxicity being correlated with the speciation forms of the heavy
metals; i.e., mixtures of HgClj and HgCli- and of CdCl+, CdCl,, and CdClj
were less toxic than the divalent cationic forms of Hg and Cd (Babich and Stotzky,
1979b, 1982~). In contrast, the clastogenicity of heavy metals appears to be
greater in the anionic than in the cationic form. For example, in FM3A cells
derived from a mouse mammary carcinoma, MnO, produced a greater number
of chromosomal aberrations than did Mn 2+; Ni(CN)i- induced a significant in-

crease, but Ni2+ only slightly increased the number of chromosomal aberrations;
and CrOi- and Cr,O:- induced large numbers of chromosomal aberrations, in-
cluding breaks and exchanges, whereas Cr3 + only produced a minor increase in
these aberrations (Umeda and Nishimura, 1979). However, the lower clastoge-
nicity of the cationic forms of the heavy metals may have resulted, as already
noted with microbial mutagenicity assays (McCann et al., 1975), from complex-
ation with organics, precipitation by PO:-, or competition by essential cations
in the tissue culture media.
The inorganic composition of various human tissues and fluids may affect the
speciation of heavy metals and, therefore, may influence their genotoxicity. For
example, the high Cl concentration in gastric juice, as a result of the secretion
of hydrochloric acid by the parietal cells, may result in the formation of metal-
Cl complexes. The inorganic ionic composition of the various body fluids and
tissues may have a differential effect on the potential genotoxicity of some heavy
metals. The concentrations of Na+, K+, CaZf, Mg2+, Cl-, HCO;, and PO:-
in plasma are 142, 4, 5, 2, 102, 26, and 2 meg/l, respectively: in interstitial fluid,
they are 145, 4, 4, 1, 116, 29, and 2 meq/l, respectively (Ruth and Patton, 1966);
whereas in urine, they are 128, 60, 5, 115, 134, 14, and 50 meq/l, respectively
(Guyton, 1971). In biological fluids and tissues, these ions may compete with
heavy metals for complexation with organics. For example, although the forma-
tion constant of the Hg-EDTA complex is high (& = 1028), EDTA is an ineffective
chelator of Hg in plasma, as a result of the competition of H+ and Ca2+ for EDTA
and of hydroxyl ions (OH-) and serum albumin for Hg2+ (Schubert, 1981).
The inorganic mineral content in the organism also appears to affect the ge-
notoxicity of heavy metals. For example, Zn protected against Cd-induced for-
mation of (1) interstitial cell tumors in mouse and rat testes (Gunn et al., 1963,
1964), (2) pleomorphic sarcomas in rats at the subcutaneous site of Cd adminis-
tration (Gunn et al., 1964), and (3) teratogenicity in chicks (Narbaitz et al., 1983)
and hamsters (Ferm and Carpenter, 1968; Ferm, 1971). Selenium (Se) protected
against Cd-induced teratogenicity in hamsters (Holmberg and Ferm, 1969).
An important factor that determines the genotoxicity of some heavy metals is
their valence, which may be influenced by the physicochemical characteristics
of the assay media (Babich et al., 1982b), of various biological fluids and tissues
(deFlora, 1978; Petrilli and deFlora, 1978a; deFlora and Boido, 1980; deFlora et
al., 1982), and of natural environments (Cranston and Murray, 1978; Babich and
Stotzky, 1980b). For example, the mediation of heavy metal-induced mutagenicity
in in vitro assays by the physicochemical characteristics that exist in the fluids
and tissues of the human body and its relevance to in vivo carcinogenicity is best
illustrated with Cr. Cr6+ is mutagenic in vitro, but its in vitro metabolic conversion
to Cr3+ eliminates its mutagenicity. Preincubation of Cr6+ in human gastric juice
or saliva presumably reduces it to Cr3+, which may explain the lack of an asso-
ciation between exposures to chromates and stomach cancer in human beings.
These reductive processes and the poor absorption of Cr3+ by the intestine appear
to be beneficial detoxification mechanisms for swallowed Cr6+. The in vitro mu-
tagenicity of Cr6+ was also decreased in the presence of lysates of human eryth-
rocytes but not in the presence of human serum or plasma. Inasmuch as Cr6+
entering the blood stream is concentrated in erythrocytes, its reduction to Cr3+
within these cells results in detoxification during transport via the blood to other
tissues and probably explains the lack of carcinogenicity of Cr compounds at a
distance from their site of administration into laboratory animals. Conversely, the
microsomal fraction from rat lung had an extremely poor inactivating effect on
the in vitro mutagenicity of Cr6+, which correlated with the selective localization
of chromate-induced tumors in the lungs of individuals exposed to Cr in the
workplace. A comparison of microsomal fractions prepared from different tissues
of Aroclor-treated rats showed the following sequence in their ability to reduce
Cr6+ to Cr3+: liver > suprarenal glands > kidney > testis > stomach > lung,
with the spleen, bladder, colon, and striated muscle showing no ability to reduce
Cr6+ to Cr3+ and, thereby, to decrease its in vitro mutagenicity. The lack of
reduction in the in vitro mutagenicity of Cr6+ by microsomal fractions from
striated muscle may explain the development of tumors at the usual site of ap-
plication of Cr6+ m . experimental animals (Petrilli and deFlora, 1978a. b, 1982.
1983; deFlora and Boido, 1980).
The oxidation-reduction potential (Et,) of the environment also can influence
the valence of some heavy metals. For example, in the oxygenated part of the
fjord of Saanich Inlet, British Columbia, Cr occurred as Cr6+, whereas in the
anoxic zone, it occurred as Cr 3+ (Cranston and Murray, 1978). Studies of the
speciation of.Cr in the Columbia River and estuary, in the Washington/Oregon
area, showed that CrOi- accounted for >90% of the dissolved Cr, with Cr3+
accounting for only about 3% of the dissolved Cr. Cr (valence undefined) also
occurred bound to particulates, with such bound Cr accounting for about 35% of
the total Cr in river samples and up to 69% in samples of estuarine waters (Cran-
ston and Murray, 1980). As chromosomal aberrations were induced in fish main-
tained in Cr6+-amended waters (Krishnaja and Rege, 1982). the effect of Eh on
the genotoxicity of Cr to the biota in aquatic environments polluted with heavy
metals requires further study. In soils, Cr occurs as Cr3+ or Cr6+ (Bartlett and
James, 1979), with interconversions between the two valences being dependent
on the E,, and the binding of the metals to organic matter (James and Bartlett,
1983a) and other soil colloids (James and Bartlett, 1983b).
In addition to being exposed to natural background levels of heavy metals,
considerable additional exposure of human beings may also occur from both
occupational and environmental pollution. Emissions from automobiles, from the
combustion of coal and oil, and from mining and other industrial activities contain
complex mixtures of heavy metals and some incompletely combusted organ&.
Interactions between these compounds occur and increase the risk of adverse
health effects. However, knowledge about these types of interactions and their
health effects is limited, although there is some information on the effects of
interactions between heavy metals in carcinogenesis. As already noted, Zn an-
tagonized the development of Cd-induced interstitial cell tumors in mouse and
rat testes and the development of pleomorphic sarcomas in rats at the subcuta-
neous site of Cd injection (Gunn et al., 1963, 1964). The carcinogenic effect of
Ni was antagonized by Mn, whereas a synergistic interaction occurred between

Ni and organic carcinogens (e.g., benzo(a)pyrene) (Nordberg and Andersen,
1981). Whereas Zn protected against Cd-induced teratogenesis in hamsters (Ferm
and Carpenter, 1967, 1968) and chicks (Narbaitz et al., 1983), Pb interacted syn-
ergistically with Cd in their teratogenicity to hamsters (Ferm, 1961).
Similarly, not much is known about the effects of interactions between multiple
heavy metals and between heavy metals and other pollutants in inducing muta-
genesis and clastogenesis. Hg 2+ did not enhance the DNA-damaging effect of
Cr6+ in the B. subtilis ret-assay (Matsui, 1980). As assayed with S. typhimurium
and E. coli, Cr6+ enhanced the mutagenicity of 9-aminoacridine and sodium azide
but not of ethyl methanesulfonate (LaVelle, 1983), and Co2+, Cu2+, Fe3+, Mn2+,
and Zn2+ enhanced the mutagenic effect of cis-.5,6-dihydro-6-hydroperoxy-5-hy-
droxythymine on transforming DNA of Hemophilus influenza (Thomas et al.,
1976). Although Cd2+ did not, whereas dimethyl nitrosamine (DMN) did, exhibit
mutagenicity in the micronucleus test using the ddY strain of mice, synergism of
mutagenicity occurred when Cd2+ and DMN were applied together (Watanabe et
al., 1982). Selinite (SeO:-) antagonized the ability of Hg2+ and methyl Hg+ to
induce sister-chromatid exchanges in human whole-blood cultures (Morimoto et
al., 1982). Cu2+, Fe3+, and Mn2+ enhanced the unscheduled DNA synthesis
induced by isoniazid and other hydrazines in cultured human fibroblasts (Whiting
et al., 1979). Benzo(u)pyrene did not affect Cr6+ -induced mutagenicity in S. ty-
phimurium, but the mutagenicity of Cr6+ was decreased by condensates from
cigarette smoke (Petrilli and deFlora, 1982). Cu2+ increased the induction of
sister-chromatid exchanges by sodium bisullite (NaHSO,) and ascorbic acid in
Chinese hamster cells (Macrae and Stich, 1979), increased the cytotoxic and
chromosome-damaging effects of ascorbic acid on human and rat cells cultured
in vitro (Stich et al., 1979), activated the mutagenic and cytotoxic effects of
ascrobic acid on S. typhimurium (Norkus et al., 1983), and activated the frag-
mentation by ascorbic acid of parental DNA in melanoma cells (Lonn and Lonn,
1983).
It is obvious from this review that to assay adequately the potential mutage-
nicity and clastogenicity and, thus, to predict accurately the potential carcino-
genicity and teratogenicity of heavy metals, it is essential that in addition to using
a battery of assays, the physicochemical characteristics of the assay media be
varied to reflect the abiotic properties of different environments in the biosphere
and the various microenvironments (i.e., tissues and fluids) of the human body.
The potential carcinogenicity and teratogenicity of heavy metals evaluated only
with standardized mutagenicity or clastogenicity assays may yield false negative
results simply because the conditions of the assays were inadequate to reflect
those in the “real world.”
CONCLUDING REMARKS
It has been estimated that 70,000 chemicals are in use and that 700 to 3000 new
chemicals will be introduced each year in the United States. The task of thor-
oughly testing all these chemicals for their potential adverse environmental and
human health effects is enormous (Hoffmann, 1982)-but obviously essential.
With the adoption of the Toxic Substances Control Act of 1976 (TSCA), the
Congress of the United States mandated the regulation, by the U.S. Environ-
mental Protection Agency (EPA), of the manufacture, processing, distribution,
commercial use, labeling, and disposal of chemicals on the basis of “unreasonable
risk” of injury to the health of human beings and to aquatic and terrestrial en-
vironments. Section 5 of TSCA requires manufacturers to submit to EPA a pre-
manufacture notice which specifies physicochemical testing and environmental
and health data before the manufacture of any “new chemical substance” or the
manufacture or processing of any existing chemical for a “significant new use.”
Congress was not specific about the types of data on environmental and health
effects that manufacturers must include in their premanufacture notice. However,
as guidance to manufacturers and to assure sufficient information for risk as-
sessments of these “new chemical substances,” EPA suggested “mutagenicity
data (screening tests),” which included studies of “gene (point) mutation,” with
the S. typhimurium reverse-mutation assay being preferred, and of “chromosome
aberrations,” with an in vitro mammalian cytotoxicity test being preferred (EPA,
1981).
These screening tests were recommended by EPA to provide information on
the genotoxicity of potential pollutants to human beings and to the biota in gen-
eral. However, the use of only standardized assay procedures greatly reduces the
reliability of these assays as accurate indicators and predictors of the potential
carcinogenicity and teratogenicity of chemicals, as the limited conditions used in
the assays may mask or inactivate the mutagenicity and clastogenicity of the
chemicals being evaluated. For example, the need of incorporating the mediating
influence of pH in genotoxicity screening assays (Leonardo and Peak, 1982), as
well as in toxicity testing (Stern and Walker, 1978; Babich and Stotzky, 1983a),
for environmental pollutants has been recognized.
The relatively few studies on the effects of physicochemical factors on the in
vitro mutagenicity of heavy metals have indicated that pH (Putrament et al.,
1975a), chelating agents (Gentile et al., 1981; Warren et al., 1981; Schultz et al.,
1982), and the valence (Nishioka, 1975) and speciation form of the heavy metals
(deFlora, 1978; Petrilli and deFlora, 1978a; deFlora and Boida, 1980; deFlora et
al., 1982) mediate their mutagenicity. Furthermore, the speciation form of heavy
metals influences their in vitro clastogenicity (Umeda and Nishimura, 1979), and
their in vivo carcinogenicity (Gunn et al., 1963, 1964) and teratogenicity (Ferm
and Carpenter, 1968; Ferm, 1971; Narbaitz et al., 1983) is influenced by the
inorganic mineral content of the test organism. These few studies, coupled with
numerous studies showing that the toxicity of heavy metals is mediated by the
various physicochemical factors of natural environments (Tables 1 and 2) and of
the human body (Table 3; Petrilli and deFlora, 1982, 1983), clearly suggest the
need to evaluate further the influence of such abiotic factors on heavy metal-
induced in vitro mutagenicity and clastogenicity and in vivo carcinogenicity and
teratogenicity.
The mutagenicity and clastogenicity of heavy metals determined only with
standardized assays may yield false negative results only because the conditions
of the assay did not reflect the actual conditions in the in vivo biologic system or
in the natural environment. Just as the in vifro and in vivo toxicities of heavy
metals to microbes and to the macrobiota are influenced by such abiotic factors
(see Babich and Stotzky, 1978, 1980a, b, 1983a, b; Stotzky and Babich, 1980,
1985a, 1985b), the mutagenicity and clastogenicity of heavy metals are undoubt-
edly also mediated by such physicochemical environmental variables.
ACKNOWLEDGMENT
This review was supported, in part, by Grant 808329 from the U.S. Environmental Protection
Agency. The views expressed in this paper are not necessarily those of the Agency.
REFERENCES
Abelson, P. H., and Aldous, E. (1950). Ion antagonism in microorganisms: Interference of normal
magnesium metabolism by nickel, cobalt. cadmium, zinc, and manganese. J. Bacreriol. 60, 401-
413.
Adelberg, E. A., Mandel, M., and Chen, G. C. C. (1965). Optimal conditions for mutagenesis by N-
methyl-N’-nitro-N-nitroguanidine in Escherichia co/i K12. Biochem. Biophys. Res. Commun. 18,
788-795.
Alink, G. M., Frederix-Welters, E. M. H.. van der Gaag, M. A., van de Kerkhoff, J. F. J., and Peels,
C. L. M. (1980). Induction of sister-chromatid exchanges in fish exposed to Rhine water. Mum.
Res. 78, 369-374.
Allen, H. E., Hall, R. H., and Brisbin, T. D. (1980). Metal speciation: Effects on aquatic toxicity.
Environ. Sri. Technol. 14, 441-443.
Amacher, D. E., and Paillet, S. C. (1980). Induction of trifluorothymidine-resistant mutants by metal
ions in L5178Y/TK+‘- cells. Mufat. Ras. 78, 279-288.
Ames, B. N. (1971). The detection of chemical mutagens with enteric bacteria. In “Chemical Muta-
gens: Principles and Methods for Their Detection” (A. Hollaender. Ed.), Vol. 1, pp. 267-282.
Plenum, New York.
Arcuri, E. J., and Ehrlich, H. L. (1977). Influence of hydrostatic pressure on the effects of the heavy
metal cations of manganese, copper, cobalt, and nickel on the growth of three deep-sea bacterial
isolates. Appl. Environ. Microbial. 33, 282-288.
Attar, E. N., and Maly, E. J. (1982). Acute toxicity of cadmium, zinc, and cadmium-zinc mixtures
to Daphnia magna. Arch. Environ. Confam. Toxicol. 11, 291-296.
Babich, H., and Stotzky, G. (1977a). Sensitivity of various bacteria, including actinomycetes, and
fungi to cadmium and the influence of pH on sensitivity. Appl. Environ. Microbial. 33, 681-695.
Babich, H., and Stotzky, G. (1977b). Effect of cadmium on fungi and on interactions between fungi
and bacteria in soil: Influence of clay minerals and pH. Appl. Environ. Microbial. 33, 10.59- 1066.
Babich, H., and Stotzky. G. (1977~). Reductions in the toxicity of cadmium to microorganisms by
clay minerals. Appl. Environ. Microbial. 33, 696-705.
Babich, H., and Stotzky, G. (1978). Effects of cadmium on the biota: Influence of environmental
factors. Adv. Appl. Micrubiol. 23, 55- 117.
Babich, H., and Stotzky, G. (1979a). Abiotic factors affecting the toxicity of lead to fungi. Appl.
Environ. Microbial. 38, 506-514.
Babich, H., and Stotzky, G. (1979b). Differential toxicities of mercury to bacteria and bacteriophages
in sea and in lake water. Canad. J. Microbial. 25, 1252-1257.
Babich, H., and Stotzky. G. (1980a). Environmental factors that influence the toxicity of heavy metals
and gaseous pollutants to microorganisms. CRC Crir. Rev. Microbial. 8, 99-145.
Babich, H., and Stotzky, G. (1980b). Physicochemical factors that affect the toxicity of heavy metals
to microbes in aquatic habitats. In “Proceedings of the ASM Conference, Aquatic Microbial
Ecology” (R. R. Colwell and J. Foster, Eds.), pp. 181-203. University of Maryland Sea Grant
Publication, College Park, Md.
Babich, H., and Stotzky, G. (1981a). Influence of water hardness on the toxicity of heavy metals to
fungi. Microbios Leff. 16, 79-84.
Babich, H., and Stotzky, G. (1981b). Components of water hardness that reduce the toxicity of nickel
to fungi. Microbios Left. 18, 17-24.
Babich, H., and Stotzky, G. (1982a). Nickel toxicity to microbes: Effect of pH and implications for
acid rain. Environ. Res. 29, 335-350.
Babich, H., and Stotzky, G. (1982b). Toxicity of nickel to microorganisms in soil: Influence of some
physicochemical characteristics. Environ. Pollut. Ser. A 29, 303-315.
Babich, H., and Stotzky, G. (1982~). Influence of chloride ions on the toxicity of cadmium to fungi.
Zentralbi. Bakteriol. Microbial. Hyg. Abt. 1, Orig. C 3, 421-426.
Babich, H., and Stotzky, G. (1982d). Nickel toxicity to fungi: Influence of some environmental factors
Ecotoxicol. Environ. Saf. 6, 577-589.
Babich, H., and Stotzky, G. (1982e). Gaseous and metal air pollutants. In “Experimental Microbial
Ecology” (R. G. Burns and J. H. Slater, Eds.), pp. 631-670. Blackwell, Oxford.
Babich, H.. and Stotzky, G. (1983,). Influence of chemical speciation on the toxicity of heavy metals
to the microbiota. in “Aquatic Toxicology” (J. 0. Nriagu. Ed.), pp. l-46. Wiley, New York.
Babich, H.. and Stotzky, G. (1983b). Toxicity of nickel to microbes: Environmental aspects. A&a.
Appl. Microbial. 29, 195-265.
Babich. H., and Stotzky. G. (1983~). Nickel toxicity to estuarineimarine fungi and its amelioration
by magnesium in sea water. Water, Air, Soil Pollut. 19, 193-202.
Babich, H., and Stotzky, Cl. (1983d). Temperature, pH, salinity. hardness. and particulates mediate
nickel toxicity to eubacteria, an actinomycete. and yeasts in lake. simulated estuarine. and sea
waters. Aquat. Toxicol. 3, 195-208.
Babich, H., and Stotzky, G. (1983e). Further studies on environmental factors that modify the toxicity
of nickel to microbes. Regal. Toxicol. Pharmacol. 3, 82-99.
Babich. H., Schiffenbauer, M., and Stotzky, G. (1982a). Comparative toxicity of trivalent and
hexavalent chromium to fungi. Bull. Environ. Contam. To.ricol. 28, 452-459.
Babich. H., Schiffenbauer, M.. and Stotzky, G. (1982b). Effects of sterilization method on the toxicity
of Crj’ and Cr6’ to fungi. Microbios Lett. 20. 55-64.
Bartlett. R.. and James, B. R. (1979). Behavior of chromium in soils: 111. Oxidation. J. Environ. Qual.
8, 31-35.
Bennicelli, C., Camoirano, A., Petruzzelli, S., Zanacchi. P., and deFlora, S. (1983). High sensitivity
of Salmonella TA102 in detecting hexavalent chromium mutagenicity and its reversal by liver and
lung preparations. Mutat. Res. 122, l-5.
Bonatti, S., Meini, M., and Abbondanolo, A. (1976). Genetic effects of potassium chromate in Schi-
zosaccharomyces pombe. Mutat. Res. 38, 147- 150.
Bridges, B. A., Mottershead, R. P., Rothwell. M. A., and Green, M. H. L. (1972). Repair-deficient
bacterial strains suitable for mutagenicity screening: Tests with the fungicide Captan. Chem.-
Biol. Interact. 5, 77-84.
Bronzetti, G., Del Carratore, R., Bauer. C., Corsi. C., Nieri, R., and Paolini, M. (1983). Detection
of genotoxicants in the leather and tannery industry using short-term tests. Bull. Environ. Contam.
Toxicol. 30, 137- 132.
Brown, E. R., Hazdra, J. J.. Keith, L., Greenspan, I., Kwapinski, J. B. G., and Beamer. P. (1973).
Frequency of fish tumors found in a polluted watershed as compared to nonpolluted Canadian
waters. Cancer Res. 33, 189-198.
Brusick. D., Gletten. F.. Jaqannath, D. R., and Weekes. U. (1976). The mutagenic activity of ferrous
sulfate for Salmonella typhimurium. Mutat. Res. 38, 386-387.
Canterford, G. S., and Canterford, D. R. (1980). Toxicity of heavy metals to the marine diatom
Ditylum brightwellii (West) Gronow: Correlation between toxicity and metal speciation. J. Mar,
Biol. Assoc. U.K. 60, 227-242.
Cantilena. L. R., and Klassen, C. D. (1981). Comparison of the effectiveness of several chelators
after single administration on the toxicity, excretion, and distribution of cadmium. Toxicol. Appl.
Pharmacol. 58, 452-460.
Cantilena, L. R., and Klassen. C. D. (1982a). The effect of repeated administration of several che-
lators on the distribution and excretion of cadmium. Toxicol. Appl. Pharmacol. 66, 361-367.
Cantilena, L. R.. and Klassen, C. D. (1982b). Decreased effectiveness of chelation therapy with time
after acute cadmium poisoning. Toxicol. Appl. Pharmacol. 63, 173-180.
Carroll, J. J., Ellis, S. J.. and Oliver. W. S. (1979). Influences of hardness constituents on the acute
toxicity of cadmium to brook trout (Salvelinus jontinalis.) Bull. Environ. Contam. Toxicol. 22,
575-581.
Casto. B. C.. Meyers. J.. and DiPaolo, J. A. (1979). Enhancement of viral transformation for evalu-
ation of the carcinogenic or mutagenic potential of inorganic metal salts. Cuncer Res. 39, 193-
198.
Casto, B. C., Pieczynski, W. J., Nelson, R. L., and DiPaolo, J. A. (1976). In vitro transformation and
enhancement of viral transformation with metals. Proc. Amer. Assoc. Cancer Res. 17, 12.
Chakoumakos, C., Russo, R. C., and Thurston, R. V. (1979). Toxicity of copper to cutthroat trout
(Salmo chzrkz) under different conditions of alkalinity, pH, and hardness. Environ. Sci. Technol.
13, 213-219.
Chapman, G., and Dunlop, S. (1981). Detoxication of zinc and cadmium by the freshwater protozoan
Tetrahymena pyriformis: 1. The effect of water hardness. Environ. Res. 26, 81-86.
Chiquoine, A. D. (1965). Effect of cadmium chloride on the pregnant albino mice. J. Reprod. Fertil.
10, 263-265.
Christie, N. T., and Costa, M. (1983). In vitro assessment of the toxicity of metal compounds. III.
Effects of metals on DNA structure and function in intact cells. Biol. Trace Elem. Res. 5,
55-71.
Coglianese, M. P. (1982). The effect of salinity on copper and silver toxicity to embryos of the Pacific
oyster. Arch. Environ. Contam. Toxicol. 11, 297-303.
Collins, Y. E., and Stotzky, G. (1982). Influence of heavy metals on the electrokinetic properties of
bacteria. Abstracts, Annu. Mtg. Amer. Sot. Microbial., p. 229.
Cranston, R. E., and Murray, J. W. (1978). The determination of chromium species in natural waters.
Anal. Chim. Acta 99, 275-282.
Cranston, R. E., and Murray, J. W. (1980). Chromium species in the Columbia River and estuary.
Limnol. Oceanogr. 25, IlO4- 1112.
deFlora, S. (1978). Metabolic deactivation of mutagens in the Salmonella-microsome test. Nature
(London) 271,455-456.
deFlora, S., and Boido, V. (1980). Effect of human gastric juice on the mutagenicity of chemicals.
Mutat. Res. 77, 307-315.
deFlora, S., Zanacchi, P., Bennicelli, C., and Arillo, A. (1982). Influence of liver S-9 preparations
from rats and rainbow trout on the activity of four mutagens. Toxicol. Lett. 10, 345-349.
Degraeve, N. (1981). Carcinogenic, teratogenic, and mutagenic effects of cadmium. Mutat. Res. 86,
115-13s.
DeKnudt, G., and Deminatti, M. (1978). Chromosome studies in human lymphocytes after in vitro
exposure to metal salts. Toxicology 10, 67-75.
DeKnudt, G., and Leonard, A. (1975). Cytogenetic investigations on leukocytes of workers from a
cadmium plant. Environ. Physiol. Biochem. 5, 319-327.
DeKnudt, G., Leonard, A., and Ivanov, B. (1973). Chromosome aberrations observed in male workers
occupationally exposed to lead. Environ. Physiol. Biochem. 3, 132-138.
DeKnudt, G., Manuel, Y., and Gerber, G. B. (1977). Chromosomal aberrations in workers profes-
sionally exposed to lead. J. Toxicol. Environ. Health 3, 885-891.
deKok, A. J., de Kniff, I?, and Mohn, G. R. (1983). Correlation between the stability of ethyl nitro-
sourea in aqueous solution at different pH and temperature and its mutagenic effectiveness for
Salmonella typhimurium strain TAlOO. Mutat. Res. 120, 81-89.
Demerec, M., Bertani, G., and Flint, J. (1951). A survey of chemicals for mutagenic action in E. co/i.
Amer. Nat. 85, 119-136.
Demerec, M., and Hanson, J. (1951). Mutagenic action of manganous chloride. Cold Spring Harbor
Symp. Quant. Biol. 16, 215-228.
Dissanayake, C. B., Tobschall, H. J., Palme. H., Rast, U., and Spettel, B. (1983). The abundance of
some major and trace elements in highly polluted sediments from the Rhine River near Mainz,
West Germany. Sci. Total Environ. 29, 243-260.
Dodge, E. E., and Theis, T. L. (1979). Effect of chemical speciation on the uptake of copper by
Chironomus tentans. Environ. Sci. Technol. 13, 1287-1288.
Donaldson, R. M., and Barreras, R. F. (1966). Intestinal absorption of trace quantities of chromium.
.I. Lab. Clin. Med. 68, 484-493.
Drasch, G. A. (1983). An increase of cadmium body burden for this century-An investigation on
human tissues. Sci. Total Environ. 26, ill- 119.
Early, J. L., and Schnell, R. C. (1981). Selenium antagonism of cadmium-induced inhibition of hepatic
drug metabolism in the male rat. Toxicol. Appl. Pharmacol. 58, 57-66.
Edgren. M., and Notter, M. (1980). Cadmium uptake by fingerlings of perch (Percafluviutilis) studied
by Cd-l 15m at two different temperatures. Bull. Environ. Confnrn. Toxicol. 24, 647-651.
Eger, M., and Doko, J. (1980). Mutagenicity and toxicity of cadmium on Saccharomyces cerevisiae.
Mutat. Res. 74, 208.
Elias, Z.. Schneider, O., Aubry, F., Daniere. M. C., and Poirot, 0. (1983). Sister chromatid exchanges
in Chinese hamster V79 cells treated with the trivalent chromium compounds chromic chloride
and chromic oxide. Curcinogenesis 4, 605-611.
Enterline, P E. (1974). Respiratory cancer among chromate workers. J. Occup. Med. 16, 523-526.
Environmental Protection Agency (1981). New chemical substances: Premanufacture testing policy.
Fed. Regist. 46, 8986-8992.
Ferm, V. H. (1971). The synteratogenic effect of lead and cadmium. Experienfiu 25, 56-57.
Ferm, V. H., and Carpenter, S. J. (1967). Teratogenic effect of cadmium and its inhibition by zinc.
Nature (London) 216, 1123.
Ferm. V. H., and Carpenter. S. J. (1968). The relationship of cadmium and zinc in experimental
mammalian teratogenesis. Lab. Invest. 18, 429-432.
Fiskesjo. G. (1971). The effects of two mercury compounds on lysogenic E. co/i K39 (X). Heredifus
69, 135-138.
Flessel, C. P. (1979). Metals as mutagenic initiators of cancer. In “Trace Metals in Health and Dis-
ease” (N. Kharasch. Ed.), pp. 109-122. Raven Press, New York.
Forsberg, C. W. (1978). Effects of heavy metals and other trace elements on the fermentative activity
of the rumen microflora and growth of functionally important rumen bacteria. Cunud. J. Microhiol.
24, 298-306.
Fox, M. R. S.. Fry, B. E. Harland, B. F., Schertel. M. E., and Weeks, C. E. (1971). Effect of ascorbic
acid on cadmium toxicity in the young coturnix. J. Nutr. 101, 1295-1306.
Frank, P. M., and Robertson. P B. (1979). The influence of salinity on toxicity of cadmium and
chromium to the blue crab, Callinectes sapidus. Bull. Environ. Sci. Technol. 21, 74-78.
Freedman, M. L.. Cunningham, P. M., Schindler. J. E., and Zimmerman, M. J. (1980). Effect of lead
speciation on toxicity. Bltll. Environ. Con&m. Toxicol. 25, 389-393.
Furst. A. (1981). Bioassay of metals for carcinogenesis: Whole animals. Environ. Heulth Perspect.
40, 83-91.
Gabbiani. G.. Gregory, A., and Bait. D. (1976). Cadmium induced selective lesions of sensory ganglia.
J. Neuroputhol. Exp. Neural. 26, 498-506.
Gale, T. E (1973). The interactions of mercury with cadmium and zinc in mammalian embryonic
development. Environ. Res. 6, 95-105.
Gasiorek, K., and Bauchinger, M. (1981). Chromosome changes in human lymphocytes after separate
and combined treatment with divalent salts of lead, cadmium, and zinc. Environ. MLltugen. 3,
513-518.
Gentile, J. M., Hyde, K., and Schubert. J. (1981). Chromium genotoxicity as influenced by complex-
ation and rate effects. Toxicol. Lett. 7, 439-448.
Gerber, G. B., Leonard, A., and Jacquet, P. (1980). Toxicity, mutagenicity, and teratogenicity of lead.
Mutut. Res. 76, 1 l5- 141.
Giesy, J. P., Leversee. G. J., and Williams, D. R. (1977). Effects of naturally occurring aquatic organic
fractions on cadmium toxicity to Simocephulus serrldutus (Daphnidae) and Gumbnsiu ufjnis
(Poeciliidae). Water Res. 11, 1013-1021.
Gipps, J. F., and Caller, B. A. W. (1982). Effect of some nutrient cations on uptake of cadmium by
Chlorella pyrenoidosa. Aust. J. Mar. Freshwater Res. 33, 979-987.
Gjessing, E. T. (1981). The effect of aquatic humus on the biological availability of cadmium. Arch.
Hydrobiol. 91, l44- 149.
Gray, S. J., and Sterling, K. (1950). Tagging of red cells and plasma proteins with radioactive chro-
mium. J. Clin. Invest. 29, 1604-1613.
Green, M. H. L.. and Muriel. W. J. (1977). Mutagen testing using Trp’ reversion in Escherichiu
co/i. In “Handbook of Mutagenicity Test Procedures” (B. J. Kilbey, M. Legator, W. Nichols,
and C. Ramel, Eds.), pp. 65-94. Elsevier, New York.
Green, M. H. L., Muriel, W. J., and Bridges, B. A. (1976). Use of a simplified fluctuation test to
detect low levels of mutagens. Mutat. Res. 38, 33-42.
Greene, J. C., Miller, W. E.. Shoroyama, T., Soltero, R. A., and Putnam, K. (1978). Use of laboratory
cultures of Selenastrum, Anabaena, and the indigenous isolate Sphaerocystis to predict effects
of nutrient and zinc interactions upon phytoplankton growth in Long Lake, Washington. Mitt.
Znt. Ver. Theor. Angew. Limnol. 21, 372-384.
Gruber. J. E., and Jennette, K. W. (1978). Metabolism of the carcinogen chromate by rat liver mi-
crosomes. Biochem. Biophys. Res. Commun. 82, 700-706.
Gunn, S. A. Gould, T. C., and Anderson, W. A. D. (1961). Zinc protection against cadmium injury
to rat testes. Arch. Pathol. 71, 52-57.
Gunn, S. A., Gould, T. C., and Anderson, W. A. D. (1963). Cadmium-induced interstitial cell tumors
in rats and mice and their prevention by zinc. J. Natl. Cancer Inst. 31, 745-749.
Gunn, S. A., Gould, T. C., and Anderson, W. A. D. (1964). Effect of zinc on carcinogenesis by
cadmium. Proc. Sot. Exp. Biol. Med. 115, 653-657.
Gunn, S. A., Gould, T. C., and Anderson, W. A. D. (1966). Protective effect of thiol compounds
against cadmium-induced vascular damage to testis. Proc. Sot. Exp. Biol. Med. 122, 1036-1039.
Gunn, S. A., Gould, T. C., and Anderson, W. A. D. (1968a). Selectivity of organ response to cadmium
injury and various protective measures. J. PathoZ. Bacterial. %, 89-96.
Gunn, S. A.. Gould, T. C., and Anderson, W. A. D. (1968b). Specificity in protection against lethality
and testicular toxicity from cadmium. Proc. Sot. Exp. Biol. Med. 128, 591-595.
Guyton, A. C. (1971). “Basic Human Physiology: Normal Function and Mechanisms of Disease.”
Saunders, Philadelphia.
Hahne, H. C. H., and Kroontje, W. (1973). Significance of pH and chloride concentration on behavior
of heavy metal pollutants: Mercury(B), cadmium(B), zinc(B), and lead(I1). J. Environ. Qual. 2,
444-450.
Harding, J. P. C., and Whitton, B. A. (1977). Environmental factors reducing the toxicity of zinc to
Stigeoclonium tenue. Brit. Phycol. 1. 12, 17-21.
Hardy, J. T., Schmidt. R. L.. and Apts, C. W. (1981). Marine sediment and interstitial water: Effects
on bioavailability of cadmium to gills on the clam Protothaca staminea. Bull. Environ. Contam.
Toxicol. 21, 798-805.
Hargreaves, J. W., and Whitton, B. A. (1976). Effect of pH on tolerance of Hormidium rivulare to
zinc and copper. Oecologia 26, 235-243.
Heck, J. D.. and Costa, M. (1982). In vitro assessment on the toxicity of metal compounds: II.
Mutagenesis. Biol. Trare Element Res. 4, 319-330.
Heinemann. B., and Howard, A. J. (1964). Induction of lambda-bacteriophages in Escherichia coli as
a screening test for potential antitumor agents. Appl. Microbiof. 12, 234-239.
Hessler, A. (1975). The effects of lead on algae. II. Mutagenesis experiments on Platymonas subcor-
diformis (Chlorophyta: Volvocales). Mutat. Res. 31, 43-47.
Hill, C. H. (1979). Studies on the ameliorating effect of ascorbic acid on mineral toxicities in the
chick. J. Nutr. 109, 84-90.
Hill, C. H., Matrone, G., Payne, W. L., and Barber, C. W. (1963). In vivo interactions of cadmium
with copper, zinc, and iron. J. Nutr. 80, 227-235.
Hoffmann, G. R. (1982). Mutagenicity testing in environmental toxicology. Environ. Sci. Technol. 16,
560A-574A.
Holmberg, R. E., and Ferm, V. H. (1969). Interrelationships of selenium, cadmium, and arsenic in
mammalian teratogenesis. Arch. Environ. Health 18, 873-877.
Hooftman, R. N., and Vink, Cl. J. (1981). Cytogenetic effects on the eastern mud-minnow, Umbra
pygmaea, exposed to ethyl methanesulfonate, benzo(a)pyrene. and river water. Ecotoxicol. En-
viron. Saf. 5, 261-269.
Houba, C., and Remacle, J. (1982). Factors influencing toxicity of cadmium to Tetrahymena pyri-
formis: particulate or soluble form and degree of complexation. Environ. Polk. Ser. A 28,
35-43.
Huisman, J., Ten Hoopen H. J. G., and Fuchs, A. (1980). The effect of temperature on the toxicity of
mercuric chloride to Scenedesmus acutus. Environ. Polk. Ser. A 22, 133-148.
Hung, Y.-W. (1982). Effect of temperature and chelating agents on cadmium uptake in the American
oyster. Bull. Environ. Contam. Toxicol. 28, 546-551.
Imreh, S., and Radulescu, D. (1982). Cytogenetic effects of chromium in vivo and in vitro. Mutat.
Res. 97, 192-193.
Jakobsen, K., and Eik-nes, K. B. (1982). Hexavalent chromium and ascorbic acid interaction on
proliferation of the human cell line NHiK3025. Toxicol. Lett. 13, 113- 118.
James, B. R., and Bartlett, R. J. (1983a). Behavior of chromium in soils: VI. Interactions between
oxidation-reduction and organic complexation. J. Environ. Qua/. 12, 173-176.
James, B. R.. and Bartlett, R. J. (1983b). Behavior of chromium in soils: VII. Adsorption and reduc-
tion of hexavalent forms. J. En\siron. Qzral. 12, 177-181.
Jennette, K. W. (1981). The role of metals in carcinogenesis: Biochemistry and metabolism. Environ.
Health Perspect. 40, 233-252.
Judy, R. D., and Davies, P. H. (1979). Effects of calcium addition as Ca(NO,)? on zinc toxicity to
fathead minnows, Pimephules promelas Rafinesque. Bull. Environ. Conram. Toxicol. 22, 88-94.
Kalinina, L. M.. and Polukhina. G. H. (1977). Mutagenic activity of heavy metal salts on Sa/monel/a
in activation systems in vivo and in vitro. Murat. Res. 46, 223-224.
Kanematsu, N., Hara, M., and Kada, T. (1980). Ret assay and mutagenicity studies on metal com-
pounds. Mutat. Res. 17, 109-116.
Kazantzis. G., and Lilly, L. J. (1979). Mutagenic and carcinogenic effects of heavy metals, In “Hand-
book on the Toxicology of Metals” (L. Friberg, G. F. Nordberg, and V. B. Vouk, Eds.). pp. 231-
272. Elsevier/North-Holland. Amsterdam.
Kiremidjian, L., and Stotzky, G. (1975). Influence of mono- and multivalent cations on the electro-
kinetic properties of Rana pipiens kidney cells. /. Cell. Physiol 85, 125-143.
Kiremidjian-Schumacher, L., and Stotzky, Cl. (1976). Influence of mono- and multivalent cations in
electrokinetic properties of normal lymphoid and Burkitt lymphoma cells. Ekperientia 32, 212-
214.
Kiremidjian-Schumacher, L., Stotzky. G.. Dickstein. R. A.. and Schwartz, J. (1981a). Influence of
cadmium. lead, and zinc on the ability of guinea pig macrophages to interact with macrophage
migration inhibitory factor. Emiron. Res. 24, 106- 116.
Kiremidjian-Schumacher, L.. Stotzky, G., Likhite. V.. Schwartz. J., and Dickstein. R. A. (1981b).
Influence of cadmium, lead, and zinc on the ability of sensitized guinea pig lymphocytes to interact
with specific antigen and to produce lymphokine. Environ. Res. 24, 96-105.
Kirsch-Volders, M. L.. Verschaeve, L., and Susanne, C. (1983). Comparative effects of HgClz and
CHsHgCl on c-mitoses and acrocentric associations in mammalian cells. Mutat. Res. 113, 269.
Knezovich, J. P., Harrison. F. L.. and Tucker. J. S. (1981). The influence of organic chelators on the
toxicity of copper to embryos of the Pacific oyster, Crassostrea gigas. Arch. Environ. Contam.
Toxicol. 10, 241-249.
Korkeala, H., and Pekkanen. T. J. (1978). The effect of pH and potassium phosphate buffer on the
toxicity of cadmium for bacteria. Acfa Vet. Scud. 19, 93- 101.
Kovalenko, T. V., and Karavaiko. G. I. (1981). Effect of temperature on the resistance of Thiobacillus
ferrooxidans to divalent copper ions. Microbiology 50, 690-695.
Krishnaja, A. I?. and Rege, M. S. (1982). Induction of chromosomal aberrations in the fish Bole-
ophthalmus dussumirri after exposure in viva to mitomycin C and heavy metals mercury, se-
lenium, and chromium. Mutat. Res. 102, 71-82.
Laborey, F., and Lavollay. J. (1977). Sur I’antitoxicite du calcium et du magnesium a I’egard du
cadmium, dans la croissance d’dspergillus niger. C.R. Acad. Sci. Ser. D. 284, 639-642.
Larramendy, M. L., Popescu, N. C.. and DiPaolo, J. A. (1981). Induction by inorganic metal salts of
sister chromatid exchanges and chromosome aberrations in human and Syrian hamster cell strains.
Environ. Mutagen. 3, 597-606.
LaValIe, J. M. (1983). Chromium/mutagen interactions in a bacterial fluctuation test. Environ. Mu-
tagen. 5, 468.
Layton, W. M.. and Ferm. V. H. (1980). Protection against cadmium-induced limb malformations by
pretreatment with cadmium or mercury. Teratology 21, 357-360.
Leonard. A., Gerber, G. B., and Jacquet, P. (1981). Carcinogenicity, mutagenicity, and teratogenicity
of nickel. Mutat. Res. 87, l-15.
Leonard, A.. Jacquet, I?, and Lauwerys, R. R. (1983). Mutagenicity and teratogenicity of mercury
compounds. Mutat. Res. 114, l-18.
Leonard, A., and Lauwerys, R. R. (1980). Carcinogenicity and mutagenicity of chromium. Mnrar,
Res. 76, 227-239.
Leonardo, J. M., and Peak, M. J. (1982). Effect of pH upon mutagenicity in Escherichin co/i. Environ.
Mutagen. 4, 335.
Levis, A. G., and Bianchi, V. (1982). Mutagenic and cytogenic effects of chromium compounds. In
“Biological and Environmental Aspects of Chromium” (S. Langard, Ed.), pp. 172-208. Elsevier,
New York.
Lofroth, G. (1978). The mutagenicity of hexavalent chromium is decreased by microsomal metabo-
lism. Naturwissenschaften 65, 207.
Lofroth, G., and Ames, B. N. (1978). Mutagenicity of inorganic compounds in Salmonella typhimu-
rium: Arsenic, chromium, and selenium. Mutat. Res. 53, 65-66.
Lonn, U., and Lonn, S. (1983). Ascorbate-Cu” fragments melanoma DNA but not fibroblast DNA
into discrete DNA populations. Curcinagenesis 4, 583-586.
Loveless, J. E., and Painter, H. A. (1968). The influence of metal ion concentrations and pH values
on the growth of a Nitrosomonas strain from activated sludge. J. Gen. Microbial. 52, I- 14.
Lower, W. R., Rose, P. S., and Drobney, V. K. (1978). In situ mutagenic and other effects associated
with lead smelting. Mutat. Res. 54, 83-93.
Lyon, T. L., and Buckman, H. 0. (1943). “The Nature and Properties of Soils.” MacMillan Co.,
New York.
Macrae, W. D., and Stich, H. F. (1979). Induction of sister chromatid exchanges in Chinese hamster
cells by the reducing agents: Bisulfite and ascorbic acid. Toxicology 13, 167-174.
Majone, F., and Levis, A. G. (1979). Chromosomal aberrations and sister-chromatid exchanges in
Chinese hamster cells treated in vitro with hexavalent chromium compounds. Mutat. Res. 67,
231-238.
Mancuso, T. F. (1980). Mortality study of beryllium industry workers’ occupational lung disease.
Environ. Res. 21, 48-55.
Martell, A. E. (1981). Chemistry of carcinogenic metals. Environ. Health Perspect. 40, 207-226.
Mason, K. E., Young, J. 0.. and Brown, J. E. (1964). Effectiveness of selenium and zinc in protecting
against cadmium-induced injury of the rat testis. And. Rec. 148, 309.
Matsui, S. (1980). Evaluation of a Bacillus subtilis ret-assay for the detection of mutagens which may
occur in water environments. Water Res. 14, 1613-1619.
McCann, J., Choi, E., Yamasaki, E., and Ames, B. N. (1975). Detection of carcinogens as mutagens
in the Salmonellaimicrosome test: Assay of 300 chemicals. Proc. Nat/. Acad. Sci. 72, 5135-
5139.
Meyer, S. A., House, W. A.. and Welch, R. M. (1982). Some metabolic interrelationships between
toxic levels of cadmium and nontoxic levels of selenium fed to rats. J. Nutr. 112, 954-961.
Michibata, H. (1981). Effect of water hardness of the toxicity of cadmium to the egg of the teleost
Oryzias latipes. Bull. En\?ron. Contam. Toxicol. 27, 187-192.
Miyaki, M., Akamatsu, N.. Ono, T., and Koyama, H. (1979). Mutagenicity of metal cations in cultured
cells from Chinese hamster. Mutat. Res. 68, 259-263.
Miyaki, M., Murata, I., Osabe, M., and Ono, T. (1977). Effect of metal cations on misincorporation
by E. co/i DNA polymerases. Biochem. Biophys. Res. Commun. 77, 854-860.
Morimoto. K., Iijima, S., and Koizumi. A. (1982). Selenite prevents the induction of sister-chromatid
exchanges by methyl mercury and mercuric chloride in human whole-blood cultures. Mutat. Res.
102, 183-192.
Muller, H.-G. (1980). Acute toxicity of potassium dichromate to Daphnia magna as a function of the
water quality. Bull. Environ. Contam. Toxicol. 25, 113-117.
Muramoto, S. (1980). Effect of complexans (EDTA, NTA, and DTPA) on the exposure to high con-
centrations of cadmium, copper, zinc, and lead. Bull. Environ. Contam. Toxicol. 25, 941-946.
Muramoto, S. (1981). Influence of complexans (EDTA, DTPA) on the toxicity of cadmium to fish at
chronic levels. Bull. Environ. Contam. Toxicol. 26, 641-646.
Murray, M. J., and Flessel, C. P. (1976). Metal-polynucleotide interactions: A comparison of carci-
nogenic and non-carcinogenic metals in vitro. Biochim. Biophys. Acta 425, 256-261.
Nakamuro. K., Yoshikawa, K.. Sayato. Y., and Kurata, H. (1978). Comparative studies on chro-
mosomal aberration and mutagenicity of trivalent and hexavalent chromium. Mutat. Res. 58,
175-181.
Narbaitz, R., Riedel, K. 0.. and Kacew, S. (1983). Induction of feather malformations in chick em-
bryos by cadmium: protection by zinc. Teratology 27, 207-213.
Nasu, Y., Kugimoto, M.. Tanaka, O., and Takimoto, A. (1983). Comparative studies on the absorption
of cadmium and copper in Lernrzu paucicosfuta. Em,iron. Polhct. Ser. A 32, 201-209.
National Academy of Sciencies (1974). “Chromium.” Nut. Acud. Sci.. Washington, D.C.
Negishi, T., and Hayatsu, H. (1980). The pH-dependent response of S&none//u typhimruium TAIOO
to mutagenic N-nitrosamines. M~fut. Res. 79, 223-230.
Nelson. D. J.. Kiremidjian-Schumacher, L.. and Stotzky. G. (1982). Effects of cadmium, lead. and
zinc on macrophage-mediated cytotoxicity toward tumor cells. Etil,irorl. Rrs. 28, 154-163.
Nestmann. E. R.. Matula. T. I.. Douglas, G. R.. Bora. K. C.. and Kowbel. D. J. (1979). Deletion of
the mutagenic activity of lead chromate using a battery of microbial tests. Mlrtc~t. Rcs. 66,
357-365.
Newbold. R. F.. Ames, J., and Cornell. J. R. (1979). The cytotoxic. mutagenic, and clastogenic effects
of chromium-containing compounds on mammalian cells in culture. M~tcrt. Res. 67, S5-63.
Nishimura. M.. and Umeda. M. (1978). Mutagenic effect of some metal compounds on cultured
mammalian cells. Mutut Res. 54, 246-247.
Nishioka, H. (1975). Mutagenic activities of compounds in bacteria. Mutut. Res. 31, l85- 189.
Nishioka, H.. and Yagi. T. (1978). Metal mutagenesis in bacteria: VI. DNA degradation. Mufut. Res.
54, 249.
Niyogi, S. K.. Feldman, R. P., and Hoffman, D. J. (1981). Selective effects of metal ions on RNA
synthesis rates. Toxicology 22, 9-21.
Nolen, G. A., Buehler, E. V., Geil, R. G., and Goldenthal, E. 1. (1972). Effects of trisodium nitrilo-
triacetate on cadmium and methyl mercury toxicity and teratogenicity in rats. Tmicol. Appl.
Pkutw~ucol 23, 122-337.
Nordberg, G. F., and Andersen, 0. (1981). Metal interactions in carcinogenesis. Enhancement, in-
hibition. Environ. Health Per-spwt. 40, 65-81.
Norkus. E. P., Kuenzig. W.. and Cooney, A. H. (1983). Studies on the mutagenic activity of ascorbic
acid in llitro and in viva. Mutut. Res. 117. 183-191.
Oberly. T. J.. Piper. C. E., and McDonald, D. S. (1982). Mutagenicity of metal salts in the L5178Y
mouse lymphoma assay. J. Toxicol. En\liron. Heulth 9, 367-376.
Ohno, H., Hanaoka, F., and Yamada. M. (1982). Inducibility of sister-chromatid exchanges by heavy
metal ions. Mutut. Res. 104, 141-145.
Onfelt. A., and Ramel. C. (1979). Some aspects on the organization of microfilaments and microtu-
bules in relation to nondisjunction. Ett\Gron. Health Perspecr. 31, 45-52.
Orgel. A.. and Orgel. L. E. (1965). Induction of mutations in bacteriophage T4 with divalent man-
ganese. J. Mol. Biol. 14, 453-457.
Parizek. J. (1957). The destructive effect of cadmium ion on testicular tissue and its prevention by
zinc. J. Endocrinol. 15, 56-63.
Parizek. J., Ostadalova, I., Benes. I., and Babicky. A. (1968). Pregnancy and trace elements: The
protective effect of compounds of an essential trace element-selenium-against the peculiar
toxic effects of cadmium during pregnancy. J. Reprod. Fert. 16, 507-509.
Paton. G. R., and Allison, A. C. (1972). Chromosome damage in human cell cultures induced by
metal salts. Mtrtut. Res. 16, 332-336.
Petrilli, F. L., and deFlora, S. (1977). Toxicity and mutagenicity of hexavalent chromium in Sulrnonellu
t~phimrrium. Appl. Environ. Microbial. 33, 805-809.
Petrilli, F. L.. and deFlora. S. (1978a). Metabolic deactivation of hexavalent chromium mutagenicity.
Mtttat. Res. 54, 139-147.
Petrilli. E L., and deFlora, S. (1978b). Oxidation of inactive trivalent chromium to the mutagenic
hexavalent form. Mutat. Res. 58, 167- 173.
Petrilli. F. L., and deFlora, S. (1982). Interpretations on chromium mutagenicity and carcinogenicity.
In “Mutagens in Our Environment” (M. Sorsa and H. Vainio. Eds.), pp. 453-464. Liss. New
York.
Petrilli. F. L., and deFlora. S. (1983). Interpretations on chromium mutagenicity and carcinogenicity.
Mutut. Res. 113, I17- 118.
Pikalek, P.. and Necasek, J. (lY83). The mutagenic activity of nickel in Couynehucrericm~ sp. Folk
Microbial. 28, 17-21.
Planas-Bohne, E, and Lehmann. M. (1983). Influence of chelating agents on the distribution and
excretion of cadmium in rats. Toxicol. Appl. PhurrnucoI. 67, 408-416.
Poldoski, J. E. (1979). Cadmium bioaccumulation assays: Their relationship to various ionic equilibria
in Lake Superior water. Environ. Sci. Technol. 13, 701-706.
Prein, A. E., Thie, G. M., Alink, G. M., and Koeman, J. H. (1978). Cytogenetic changes in fish
exposed to water of the river Rhine. Sci. Total Environ. 9, 287-291.
Putrament, A., Baranowska, H., and Prazmo, W. (1973). Induction by manganese of mitochondrial
antibiotic resistance mutations in yeast. Mol. Gen. Genet. 126, 357-366.
Putrament, A., Baranowska. H., Ejchart, A., and Prazmo, W. (1975a). Manganese mutagenesis in
yeast: A practical application of manganese for the induction of mitochondrial antibiotic-resistant
mutations. J. Cert. Microbial. 62, 265-270.
Putrament. A., Baranowska, H.. Ejchart, A., and Prazmo, W. (1975b). Manganese mutagenesis in
yeast: IV. The effects of magnesium, protein synthesis inhibitors, and hydroxyurea on AntR
induction in mitochondrial DNA. Mol. Gen. Getter. 140, 339-347.
Putrament, A., Baranowska. H., Ejchart, A.. and Jachymczyk, W. (1977). Manganese mutagenesis
in yeast: VI. Mn?+ uptake, mitDNA replication, and ER induction. Comparison with other di-
valent cations. Mol. Gen. Genet. 151, 69-76.
Rai, L. C., Gaur, J. P., and Kumar, H. D. (1981). Protective effects of certain environmental factors
on the toxicity of zinc, mercury, and methylmercury to Chlorella vulgaris. Environ. Res. 25,
250-259.
Ramamoorthy, S., and Kushner, D. J. (1975). Binding of mercuric and other heavy metal ions by
microbial growth media. Microb. Ecol. 2, 162-176.
Reashid. M. A. (1971). Role of humic acids of marine origin and their different molecular weight
fractions in complexing di- and tri-valent metals. Soil Sci. 111, 298-306.
Reid, G. K. (1961). “Ecology of Inland Waters and Estuaries.” Reinhold, New York.
Rohr, G., and Bauchinger, M. (1976). Chromosome analyses in cell cultures of the Chinese hamster
after application of cadmium sulphate. Mutat. Res. 40, 125-130.
Rosenkranz, H. S., and Poirier, L. A. (1979). Evaluation of the mutagenicity and DNA modifying
activity of carcinogens and noncarcinogens in microbial systems. J. Natl. Cancer Inst. 62, 873-
891.
Rossman, T. G. (1981). Effect of metals on mutagenesis and DNA repair. Environ. Health Perspect.
40, 189-195.
Ruth, T. C., and Patton, H. D. (1966). “Physiology and Biophysics.” Saunders, Philadelphia.
Rudd. T., Sterritt, R. M., and Lester, J. N. (1983). Mass balance of heavy metal uptake by encap-
sulated cultures of Klebsiella aerogenes. Microb. Ecol. 9, 261-272.
Sakaguchi, T., Tsuji. T., Nakajima, A., and Horikoshi, T. (1979). Accumulation of cadmium by green
microalgae. Eur. J. Appl. Microbial. Biotechnol. 8, 207-215.
Sarto, F., Caominato, I., Bianchi. V., and Levis, A. G. (1982). Increased incidence of chromosomal
aberrations and sister chromatid exchanges in workers exposed to chromic acid (CrO,) in elec-
troplating factories. Carcinogenesis 3, 101 l-1016.
Say, P. J., and Whitton, B. A. (1977). Influence of zinc on lotic plants: II. Environmental effects on
toxicity of zinc to Hormidium rivulare. Freshwater Biol. 7, 377-384.
Scharpf, L. G., Hill, I. D., Wright, P. L., Plank, J. B., Keplinger, M. L., and Calandra, J. C. (1972).
Effect of sodium nitrilotriacetate on toxicity, teratogenicity, and tissue distribution of cadmium.
Nature (London) 239, 23 l-234.
Schmid, E., Bauchinger. M., Pietruck, S., and Hall, G. (1972). Die cytogenetische wirkung von blei
in menschlichen peripheren lymphocyten in vitro und in vivo. Mutat. Res. 16, 401-406.
Schubert, J. (1981). Chelating agents in biological systems. Environ. Health Perspect. 40, 227-232.
Schultz, P. N.. Warren, G., Kosso. C.. and Rogers, S. (1982). Mutagenicity of a series of hexacoor-
dinate cobalt(II1) compounds. Mutat. Res. 102, 393-400.
Shehata, F. H. A., and Whitton, B. A. (1982). Zinc tolerances in strains of the blue-green alga Ana-
cystis nidulans. Brit. Phycol. J. 17, 5-12.
Shirasu, Y., Moriya, A., Kato, K., Furuhashi, A., and Kada, T. (1976). Mutagenicity screening of
pesticides in the microbial system. Mutat. Res. 40, 19-30.
Sidereis, E. G., and Petraki, H. (1982). Sister-chromatid exchanges and chromosome structure.
Mutat. Res. 97, 222.
Simmon, V. F. (1979). In vitro mutagenicity assays of chemical carcinogens and related compounds
with Salmonella typhimurium. J. Natl. Cancer Inst. 62, 893-899.
Singh, C., and Kaul. B. L. (1978). Role of pH in chemical mutagenesis. J. Sci. Ind. Res. 37, 4?6-
432.
Singh, I. (1983). Induction of reverse mutation and mitotic gene conversion by some metal compounds
in Saccharomyces cerevisiae. Mutat. Res. 117, 149-152.
Singh, S. P., and Kashyap, A. K. (1978). Manganese toxicity and mutagenesis in two blue-green algae.
Environ. Exp. Bot. 18, 47-53.
Singh, S. P., and Pandey, A. K. (1981). Cadmium toxicity in a cyanobacterium: Effect of modifying
factors. Emiron. Exp. Bot. 21, 257-265.
Sirover, M. A., and Loeb. L. A. (1976). Infidelity of DNA synthesis in vitro: Screening for potential
mutagens or carcinogens. Science (Washington. D.C.) 194, 1434-1436.
Sirover, M. A., and Loeb. L. A. (1977). On the fidelity of DNA replication. J. Biol. Chem. 252,
3605-3610.
Smith, M. J., and Heath, A. G. (1979). Acute toxicity of copper. chromate, zinc, and cyanide to
freshwater fish: Effect of different temperatures. Bull. Environ. Conrum. Toxicol. 22, 113-l 19.
Spencer, D. F.. and Nichols, L. H. (1983). Free nickel ion inhibits growth of two species of green
algae. Environ. Pollut. Ser. A 31, 97-104.
Steemann Nielsen, E., and Kamp-Nielsen, L. (1970). Influence of deleterious concentrations of
copper on the growth of Chlorella pyrenoidosa. Physiol. Plant. 23, 828-840.
Stella, M., Montaldi, A.. Rossim R., Rossi, G.. and Levis, A. G. (1982). Clastogenic effects of
chromium on human lymphocytes in vitro and in vi~ao. Mutat. Res. 101, 151- 164.
Stendahl, D. H., and Sprague, J. B. (1982). Effects of water hardness and pH on vanadium lethality
to rainbow trout. Water Res. 16, 1479-1488.
Stephenson, R. R. (1983). Effects of water hardness, water temperature, and size of the test organism
on the susceptibility on the freshwater shrimp, Gummarus pulex (L.). to toxicants. Bull. Emjiron.
Contam. Toxicol. 31, 459-466.
Stern, A. M., and Walker. C. R. (1978). Hazard assessment of toxic substances: Environmental fate
testing of organic chemicals and ecological effects testing. In “Estimating the Hazard of Chemical
Substances to Aquatic Life” (J. Cairns, K. L. Dickson, and A. W. Maki. Eds.). pp. 81-131.
Amer. Sot. Testing Materials. Philadelphia.
Stevenson, F. J. (1972). Role and function on humus in soil with emphasis on adsorption of herbicides
and chelation of micronutrients. BioScience 22, 643-652.
Stich, H. F.. Karim, J.. Koropatnick, J., and Lo, L. (1976). Mutagenic action of ascorbic acid. Natrrre
(London) 260, 722-724.
Stich, H. F.. Wei, L., and Whiting, R. F. (1979). The mutagenic and anti-mutagenic effects of ascor-
bate and ascorbate-metal mixtures. Emjiron. Mutugen. 1, 119.
Stotzky. G. (1972). Activity. ecology, and population dynamics of microorganisms in soil. CRC Crir.
Rel’. Microbial. 2, 59- 137.
Stotzky, G.. and Babich, H. (1980). Mediation of the toxicity of pollutants to microbes by the phys-
icochemical composition of the recipient environment, In “Microbiology-1980” (D. Schlessinger,
Ed.). pp. 353-354. Amer. Sot. Microbial., Washington, D.C.
Stotzky, G.. and Babich, H. (1985a). Physicochemical environmental factors influence the toxicity of
heavy metals to microbes. In “Annales d’universite de Nancy.” Nancy, France (in press).
Stotzky, G.. and Babich. H. (1985b). Physicochemical environmental factors affect the response of
microorganisms to heavy metals: Implications for the application of microbiology to mineral
exploration. In ‘*Organic Matter, Biological Systems, and Mineral Exploration” (D. Carlisle.
Ed.). Prentice-Hall, Englewood Cliff, New Jersey (in press).
Strand, F. (1983). “Physiology. A Regulatory Systems Approach.” MacMillan Co., New York.
Sugawara, N., Aoshima. K.. and Kasuya, M. (1983). Effect of cadmium chloride and Cd-metallothi-
onein on the nervous tissue culture. Toxicol. Lett. 16, 95-101.
Sunda. W., and Guillard, R. R. C. (1976). The relationship between cupric ion activity and the toxicity
of copper to phytoplankton. J. Mar. Res. 34, 51 l-529.
Sunderman. F. W. (1978). Carcinogenic effects of metals. Fed. Proc. 37, 40-46.
Sunderman, F. W. (1979). Mechanisms of metal carcinogenesis. Biol. Truce Elem. Res. 1, 63-86.
Szeto, C., and Nyberg. D. (1979). The effect of temperature on copper tolerance of Paramecium.
Bull. Environ. Contam. Toxicol. 21.131-135.
Tait, R. V., and desanto, R. S. (1972). “Elements of Marine Ecology.” Springer-Verlag, New York/
Berlin.
Thomas, H. F., Herriott, R. M., Hahn, B. S., and Wang, S. Y. (1976). Thymine hydroperoxide as a
mediator in ionizing radiation mutagenesis. Nature (London) 259, 341-342.
Tindall, K. R., Warren, G. R., and Skaar. P. D. (1978). Metal ion effects in microbial screening
systems. Mutat. Res. 53, 90-91.
Tsapakos, M. J., and Wetterhahn, K. E. (1983). The interaction of chromium with nucleic acids.
Chern-Biol. Interact. 46, 265-277.
Tso, W.-W., and Fung, W.-P. (1981). Mutagenicity of metallic cations. Toxicol. Lett. 8, 195-200.
Tsuda, T., and Kato, K. (1977). Chromosomal aberrations and morphological transformation in ham-
ster embryonic cells treated with potassium dichromate in vitro. Mutat. Res. 46, 87-94.
Umeda, M., and Nishimura, M. (1979). Inducibility of chromosomal aberrations by metal compounds
in cultured mammalian cells. Mutat. Res. 67, 221-229.
Vainio, H., and Sorsa, M. (1981). Chromosome aberrations and their relevance to metal carcinogen-
esis. Environ. Health Perspect. 40, 173-180.
van Kreijl, Kool, C. F., de Vries. M., van Kranen, H. J., and de Greef. E. (1980). Mutagenic activity
in the rivers Rhine and Meuse in the Netherlands. Sci. Total Environ. 15, 137-147.
Venier, P., Montaldi, A., Majone, F.. Bianchi, V., Sarto, F., and Levis, A. G. (1982). Cytotoxic,
mutagenic, and clastogenic effects of industrial chromium compounds. Carcinogenesis 3, 1331-
1338.
Venier, P., Montaldi, A., Majone, F., Bianchi, V., Sarto, F., and Levis, A. G. (1983). Cytotoxic,
mutagenic, and clastogenic activity of industrial chromium compounds. Mufut. Res. 113, 157-
158.
Venitt, S., and Levy, L. S. (1974). Mutagenicity of chromates in bacteria and its relevance to chromate
carcinogenesis. Nature (London) 250, 493-495.
Vielmetter, W., and Schuster, H. (1960). The base specificity of mutation induced by nitrous acid in
phage T2. Biochem. Biophys. Res. Commun. 2, 324-328.
Wagoner, J. K., Infante. P. F., and Bayliss, D. L. (1980). Beryllium: an etiologic agent in the induction
of lung cancer, nonneoplastic respiratory disease, and heart disease among industrially exposed
workers. Environ. Res. 21, 15-34.
Warren, G., Schultz, P., Bancroft, D., Bennett, K., Abbott, E. H.. and Rogers. S. (1981). Mutage-
nicity of a series of hexacoordinate chromium(III) compounds. Mutat. Res. 90, 11l- 118.
Watanabe, M., Honda, S., Hayashi. M., and Matsuda, T. (1982). Mutagenic effects of combinations
of chemical carcinogens and environmental pollutants in mice as shown by the micronucleus test.
Mutat. Res. 97, 43-48.
Webster, W. S. (1979). Cadmium-induced fetal growth retardation in mice and the effects of dietary
supplements of zinc, copper, iron, and selenium. J. Nutr. 109, 1646-1651.
Weed, L. (1963). Effects of copper on Bacillus subtilis J. Bacterial. 85, 1003-1010.
Whiting, R. F., Wei, L., and Stich, H. F. (1979). Enhancement by transition metals of unscheduled
DNA synthesis induced by isoniazid and related hydrazines in cultured normal and xeroderma
pigmentosum human cells. Mutat. Res. 62, 505-515.
Wilson, D. E. (1978). An equilibrium model describing the influence of humic materials on the spe-
ciation of Cu*+, Zn*+, and Mn*+ in freshwaters. Limnol. Oceanogr. 23, 499-507.
Wong, K. H., Chan, K. Y., and Ng, S. L. (1979). Cadmium uptake by the unicellular green alga
Chlorella salinu Cu-1 from culture media with high salinity. Chemosphere 8, 887-891.
Wong, M. H. (1980). Toxic effects of cobalt and zinc to Chlorella pyrenoidosa (26) in soft and hard
water. Microbios 28, 19-25.
Wright, D. A., and Frain, J. W. (1981). The effect of calcium on cadmium toxicity in the freshwater
amphipod, Gammarus pulex (L.). Arch. Environ. Contam. Toxicol. 10, 321-328.
Yunice, A., and Perry, H. M. (1961). The acute pressor effect of parenteral cadmium and mercury
ions. .I. Lab. C/in. Med. 58, 975.
Zakour. R. A., Kundel, T. A., and Loeb, L. A. (1981). Metal-induced infidelity of DNA synthesis.
Environ. Health Perspect. 40, 197-205.
Zasukhina, G. D., Sineschikova, T. A., Llova, G. N., and Kirkova, Z. S. (1977). Molecular-genetic
effects of cadmium chloride. Mutat. Res. 45, 169-174.

ảnh hưởng của kln

Uploaded by

Copyright:

Available Formats

ảnh hưởng của kln

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

ảnh hưởng của kln

Uploaded by

Copyright:

Available Formats

ENVIRONMENTAL RESEARCH 37, 253-286 (1985)

PH Increasing the pH from acidic to alkaline levels reduced the toxicity of

PH Increasing the pH from 4.1 to 5.1 increased the toxicity of Cd and Cu to

cells (Kiremidjian and Stotzky, 1975; Kiremidjian-Schumacher and Stotzky. 1976;

Cell line Comments and references

Biologic system Results References

Biologic system Results References

EFFECT OF PHYSICOCHEMICAL FACTORS ON THE MUTAGENICITY AND

charomyces cerevisiae was pH dependent, as increasing the pH from 6 to 6.7

sodium chloride adjusted to pH 1.3 resulted in 890 ? 42 and 823 * 37 revertants,

The genotoxicity of heavy metals is apparently not necessarily decreased by

of chromosomal aberrations than did Mn 2+; Ni(CN)i- induced a significant in-

Ni was antagonized by Mn, whereas a synergistic interaction occurred between

You might also like