Biol Fertil Soils (1992) 13:181-186
BiologyandFert~ty
Simultaneous estimates of the diversity and the degradative capability
of heavy-metal-affected soil bacterial communities*
H . H . Reber
Institut far Bodenbiologie, Bundesforschungsanstalt ftir Landwirtschaft, BundesaIIee 50, W-3300 Braunschweig, Federal Republic of Germany
Received July 31, 1991
Summary. A method for simultaneous estimation of the
impact of heavy metal stress on the diversity and the
degradative capability of soil bacteria was developed and
tested. It is based on the ability of soil bacteria to use aro-
matic substances as C sources. Though these characters
were selected to indicate specific biochemical potential,
they were sufficiently capable of differentiating the iso-
lated strains into biochemical types. Using these charac-
ters implied that only organisms capable o f growing at
the expense of aromatics were tested. However, this made
it possible to restrict the number of assays to 20 and to
test up to 200 isolates per soil sample. In three out of five
experiments, we found that heavy metal stress definitely
decreased the diversity of bacteria in a flora. In two other
experiments, an unchanged or even higher diversity in the
metal-contaminated soils was observed. These unexpect-
ed results may have been caused by a high soil pH render-
ing metals unavailable or by selection of fast-growing
strains in the control soil (decrease in evenness). The
relative scoring of all characters in a community (also the
average number of substrates used per isolate) was not a
reliable indicator of changes in the degradative capability
of bacterial communities. However, the ratio of the 10
lowest-scoring to the 10 highest-scoring out of the 20
characters tested was capable of indicating these changes.
In all heavy-metal-affected bacterial communities so far
tested, this ratio was lower than in the corresponding un-
affected communities. These data suggest that some of
the rare biochemical capabilities of bacterial flora were
lost following contamination of soils by heavy metals.
Key words: Heavy metals - Soil bacterial community -
Diversity - Biochemical capability - Aromatic sub-
strates - Cluster analysis - Rarefaction
Investigations on the impact of heavy metals on soil
microflora have so far concentrated on quantitative as-
* Dedicated to the memory of my son Ulrich
~
9 Springer-Verlag 1992
pects that demonstrate decreases in soil microbial num-
bers or biomass (Bisessar 1982; Brookes and McGrath
1984) and on specific or non-specific soil activities such
as respiration, dehydrogenase, or N2 fixation (Doelman
and Haanstra 1979, 1986; Chang and Broadbent 1981;
Tyler 1981; Brookes et al. 1986; Reber 1989). Much less
is known about the impact of heavy metals on the diversi-
ty of bacterial communities. In a methodological review,
RosswaI1 and Kvillner (1978) cited only one paper (de
Leval et al. 1976) dealing with the effect of heavy metals
on aquatic bacteria. Atlas (1984), reviewing the impact of
environmental stress on microbial diversity, apparently
did not find any relevant paper on heavy metal effects on
the community structure of soil bacteria. Barkay et al.
(1985) were the first to study the diversity of bacteria in
metal-contaminated soils. The reason why there have
been so few soil microfloristic analyses in heavy metal mi-
crobiology is certainly the time and labor involved in this
methodology. However, Mills and Wassel (1980) were able
to demonstrate that the diversity of microorganisms can
satisfactorily be described by as little as 20 differentiating
characters, making it possible to increase the number of
microbial isolates that can be tested.
The aim of the present investigation was to develop a
method which, in addition to giving sufficient discrimi-
nation between isolates, would also provide data on the
degradative ability of a soil bacterial community. Assess-
ment of the ability to grow with 20, mostly aromatic, C
sources was expected to serve both purposes. Aromatics
are very diverse in structure, their catabolism requiring
specific enzymes and leading to a few central aromatic in-
termediates before the ring structure is lost (Dagley and
Nicholsen 1970). Consequently, only bacteria capable o f
growing with these substrates were analysed in this study.
Materials and methods
Soils
Physical, chemical,and biologicalproperties of the soils used in this in-
vestigation are shown in Tablel. The soil used in experiment 1 was an