Biol Fertil Soils (1994) 17:177-184
R Doelman 9 E. Jansen 9 M. Michels 9 M. van Til
Effects of heavy metals in soil on microbial diversity
and activity as shown by the sensitivity-resistance index,
an ecologically relevant parameter
Received: 15 April 1993
Abstract A sensitivity-resistance index was developed,
and proved to be a very sensitive biomonitor of soil pollu-
tion with heavy metals. The index was developed by a
step-by-step approach. Ultimately, the bacterial soil mi-
croflora was divided into three groups, sensitive, tolerant,
and resistant microflora. Zn and Cd sensitivity was de-
fined as no growth occurring in the presence of 5 and
0.5 mg 1-1 of these metals, respectively, while resistance
was defined as distinct growth in the presence of 50 and
16 mg 1-1, respectively. The sensitivity: resistance ratio
of a referent clay soil ( 0 . 5 7 m g C d k g -1 and 140mg
Z n k g -1 was 0.53, but for polluted ( 6 m g C d k g - l +
670mg Zn kg -1) clay soil, the ratio was 0.24. For a
referent (0.06 mg Cd kg -1 + 12 mg Zn kg -1) sandy soil
the sensitivity: resistance ratio was 1.50 whereas polluted
(2.3 mg Cd kg -1 +252 mg Zn kg -1) sandy soil had a ratio
0.19. The ecological value of the sensitivity-resistance lies
in its capacity to reflect potential degradation of aromatic
compounds. It has been shown repeatedly that sensitive
bacteria grow significantly better on a range of selected
aromatic compounds. It has been speculated that resis-
tance fo heavy metals may reduce the bioremediation ca-
pacity of soil towards chlorinated aromatics and poly-
aromatic hydrocarbons.
Key words Heavy metals 9 Microbial activity 9 Microbial
diversity 9 Sensitivity-resistance index 9 Biomonitoring
Introduction
Because of their function in recycling elements, their
ubiquity, their abundance, their shape and small size and,
E. Jansen (~) 9 M. Michels 9 M. van Til
Department of Ecotoxicology of the Institute for Forestry
and Nature Research, P.O. Box 23,
6700 AA Wageningen, The Netherlands
P. Doelman
IWACO BY., Consultants for water and environment, P.O. Box 8520,
3009 AM Rotterdam, The Netherlands
9 Springer-Verlag 1994
consequently, extremely high surface area to volume ratio,
microorganisms are the first group of soil organisms to be
affected by diffuse soil pollution. However, it is extremely
difficult to use the entire microbial community in soil as
a monitor for heavy metal pollution. The heterogeneity of
soil on a small scale, particularly in factors such as organ-
ic matter content, pH, and the silt fraction, and the varia-
tion in temperature, water and oxygen content, and nutri-
ent supply lead to extreme natural fluctuations; in micro-
bial activity and numbers. Even clearly defined features
and omnipresent levels such as the adenylate energy
charge (Brooks and McGrath 1987) are not ,;uitable as
yardsticks for heavy metal pollution. An accumulation of
organic matter in soil is the first indication, visible to the
naked eye, that pollution of heavy metals affects the de-
composer microbial community. This has been known for
more than 20 years (Kendrick 1962; Pugh and Williams
1971; Rtihling and Tyler 1973). Other, less visible but
microbiologically distinct, effects include the inhibition
of specific steps in the C, N, P, and S cycles.
For respiration, urease, phosphatase, and arylsul-
phatase activity, the decrease in resistance proceeds ac-
cording to a logistic dose-response curve, as indicated by
Haanstra et al. (1985) and shown in Fig. 1. Before a sig-
nificant adverse dose-activity response can be detected, it
appears that a qualitative shift occurs within the entire
contributing community; similar activities are carried out
by other, less sensitive, organisms. However, there is little
information on the effects of heavy metals on numbers
and versatility of so-called sensitive and resistant soil mi-
croorganisms.
Prokaryotes (bacteria, actinomycetes) are more sensi-
tive than eukaryotes (fungi) to heavy metal pollution of
soil. Bacteria are more sensitive than actinomycetes, and
Gram-positive bacteria are more sensitive than Gram-neg-
ative bacteria (Babich and Stotzky 1983; Doelraan 1985).
Doelman and Haanstra (1979) showed that 7:5~ of the
bacteria isolated from Pb-polluted soils ( 1 8 4 - 1 1 7 7 m g
Pb kg -1) were able to grow on a medium containing
30 gg Pb m1-1, whereas only 5~ of bacteria from refer-
ence soils ( 1 2 - 4 1 mg Pb kg -1) were able to grow on the