See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/248891464

Soil microbial biomass—what do the numbers really mean?

Article  in  Australian Journal of Experimental Agriculture · January 1998

DOI: 10.1071/EA97142

CITATIONS READS

211 548

1 author:

Ram C Dalal
Queensland Government
282 PUBLICATIONS   8,741 CITATIONS   

SEE PROFILE

Some of the authors of this publication are also working on these related projects:

article View project

Strategic tillage View project

All content following this page was uploaded by Ram C Dalal on 14 April 2016.

The user has requested enhancement of the downloaded file.

C S I R O P U B L I S H I N G

Australian Journal
of Experimental Agriculture
Volume 38, 1998
© CSIRO 1998

… a journal publishing papers (in the soil, plant and animal sciences)
at the cutting edge of applied agricultural research

w w w. p u b l i s h . c s i r o . a u / j o u r n a l s / a j e a
All enquiries and manuscripts should be directed to
Australian Journal of Experimental Agriculture
CSIRO PUBLISHING
PO Box 1139 (150 Oxford St)
Collingwood
Vic. 3066 Published by
Australia
CSIRO PUBLISHING
Telephone: 61 3 9662 7614 in co-operation with the
Facsimile: 61 3 9662 7611
Email: chris.anderson@publish.csiro.au Standing Committee on Agriculture
lalina.muir@publish.csiro.au and Resource Management (SCARM)
Australian Journal of Experimental Agriculture, 1998, 38, 649–65
Soil microbial biomass—what do the numbers really
mean?
R. C. Dalal
Department of Natural Resources, QWRI and RSC, 80 Meiers Road, Indooroopilly, Qld 4068, Australia;
e-mail: Ram.Dalal@dnr.qld.gov.au
Summary. Soil microbial biomass comprises less than
5% of organic matter in soil. However, it performs at
least 3 critical functions in soil and the environment. It
is a labile source of carbon, nitrogen, phosphorus, and
sulfur; it is an immediate sink of carbon, nitrogen,
phosphorus and sulfur; and it is an agent of nutrient
transformation and pesticide degradation. In addition,
microorganisms form symbiotic associations with roots,
act as biological agents against plant pathogens,
contribute towards soil aggregation, and participate in
soil formation.
Critical evaluation of the significance of soil
microbial biomass is hampered by the reliable
measurement of microbial biomass, and simultaneous
partitioning of its 3 major functions in soil. For
comparative purposes, soil microbial biomass and its
derived indices have been successfully used to measure
Introduction
Soil microbial biomass is the living component of
soil organic matter. It excludes soil animals and plant
roots. Although it comprises less than 5% of organic
matter in soil, it performs at least 3 critical functions
for plant production in the ecosystem. It is a labile
source of carbon (C), nitrogen (N), phosphorus (P), and
sulfur (S); it is an immediate sink of C, N, P and S;
and it is an agent of nutrient transformation and
pesticide degradation. In addition, microorganisms form
symbiotic associations with roots, act as biological
agents against plant pathogens, contribute towards soil
aggregation (Angers et al. 1992), and participate in soil
formation.
In the past, interest in microbial biomass assessment
in soil has been limited by tedious, time consuming and
unreliable techniques (direct microscopy, culture media)
for enumeration of individual microbial communities.
Estimation of microbial biomass from actual
enumeration of the whole suite of microorganisms
(e.g. bacteria, fungi, actinomycetes, protozoa,
© CSIRO 1998
649
early changes induced by land use practices, zero
tillage, crop rotations and other cultural practices,
nutrient cycling, land disposal of sewage sludge, and
applications of herbicides and insecticides. However, as
a routine analytical tool, it is limited by the
cumbersome and time consuming measurements, lack
of benchmarking values and interpretation, ambiguous
relationship with productivity, and cost-effectiveness.
With increasing demand to monitor soil quality and
protection of the environment, improved and rapid
techniques, including molecular biotechniques, will be
required to measure soil microbial biomass for its size
of sink, source and rates of turnover. Eventually,
agricultural science will benefit and utilise soil
microbial biomass as an analytical tool to produce
abundant, economical, and clean food and fibre.
nematodes) has been difficult because of incomplete
extraction, inappropriate growth media, extremely
variable growth habits of microorganisms in soil, and
scant information on their biovolumes (Jenkinson 1988).
In the last 20 years, relatively rapid assessment of soil
microbial biomass has been possible based on
physiological, biochemical and chemical techniques
(Horwath and Paul 1994). These include chloroform
fumigation incubation (CFI) (Jenkinson and Powlson
1976), chloroform fumigation extraction (CFE) (Brookes
et al. 1985; Vance et al. 1987), substrate-induced
respiration (SIR) (Anderson and Domsch 1978), and
adenosine triphosphate (ATP) analysis (Jenkinson et al.
1979; Eiland 1983; Webster et al. 1984). Of these, the
first 2 methods have been widely used to estimate
microbial biomass in agricultural, pastoral and forestry
systems, rehabilitation of disturbed lands, and pesticide
and heavy metals polluted lands and materials. Recently,
microbial biomass has even been proposed as a sensitive
indicator of soil quality (Karlen et al. 1997) and soil
health (Sparling 1997).
10.1071/EA97142
650
But what do the soil microbial biomass numbers
really mean? I will examine this question and seek
answers in the following sequence: (i) methods and their
limitations to estimate microbial biomass; (ii) effects of
environment, soil, cultural practices and additives on the
size of soil microbial biomass; (iii) source of nutrients
and rate of mineralisation and its relationship to crop
requirements; (iv) nutrient sink; (v) pesticide
degradation; and (vi) as an indicator of soil quality.
Methods to estimate soil microbial biomass
Detailed descriptions of methods and their limitations
to estimate soil microbial biomass by CFI, CFE, SIR and
ATP analysis are given by Jenkinson (1988), Horwath
and Paul (1994) and Martens (1995). Both CFI and CFE
methods measure a standing crop of soil microbial
biomass, while SIR essentially measures microbial
activity and ATP is an indicator of microbial dynamics in
soil. Chloroform fumigation incubation and CFE can
also be used to estimate N, P and S in soil microbial
biomass and these can usually be used to recover added
radiotracers (e.g. 14C, 15N, 32P and 35S) from microbial
biomass.
The microbial biomass of individual groups such as
bacteria and fungi is estimated by microscopic methods
and by measuring specific substances. The microbial
cell-wall components, muramic acid and glucosamine
have been used as indicators of bacterial and fungal
biomass respectively (Atlas and Bartha 1981). However,
microscopic techniques, and muramic acid and
glucosamine methods have difficulties in distinguishing
between living and non-living microbial biomass. The
cell-membrane-bound components such as phospholipid
fatty acids, have been used to estimate fungal and
bacterial biomass in soil (Zelles et al. 1992; Frostegard
and Baath 1996).
Only a brief description and limitations of these
methods are given below.
Chloroform fumigation incubation
In this method, a moist soil is fumigated with ethanol-
free chloroform for 24 h; chloroform is then removed by
repeated evacuation; the soil is reinoculated with a small
amount of unfumigated soil and then incubated at a
constant temperature (usually 22 or 25oC) for 10 days at
field capacity or 50% of its water holding capacity
(about –0.01 MPa). An additional soil sample is retained
unfumigated and used as a control. The CO2 evolved
during incubation can be measured by gas
chromatography, as a continuous flow or by sorption in
alkali followed by titrimetric, conductometric or
colorimetric determination. As the net C mineralised as
CO2 is only a proportion of the total microbial biomass
C, a kC factor is used to calculate total soil biomass C.
For soil microbial biomass N determination, mineral
N (NH 4 -N and NO 3 -N) from both fumigated and
R. C. Dalal
unfumigated (control) samples are extracted with 2 mol
KCl/L after incubation. The mineral N in the extracts is
then determined colorimetrically or by steam distillation.
As for microbial biomass C, a kN factor is used to correct
for incomplete mineralisation of N from killed
microorganisms for calculating total biomass N.
Soil microbial biomass C and N are calculated from
equations (1) and (2):
Biomass C = (CO2-C fumigated – CO2-C control)/kC (1)
Biomass N = (mineral N fumigated – mineral N control)/kN (2)
The widely accepted k C value is 0.41 at 22 o C
(Anderson and Domsch 1978) or 0.45 at 25 o C
(Jenkinson and Powlson 1976). However, kN varies from
0.30 to 0.68 (Smith and Paul 1990). Jenkinson (1988)
suggested a kN value of 0.57 at 25oC, which is about
0.50 at 22oC.
Two basic assumptions of the CFI method are: (i) that
CO2-C evolved or mineral N produced during incubation
in fumigated soil must exceed that from the
corresponding unfumigated soil; and (ii) that CO 2-C
evolved or mineral N produced during incubation from
the non-microbial source must be equal in both
fumigated and unfumigated soil samples (Jenkinson
1988).
In soils with relatively low microbial biomass but
high respiration activity, subtraction of the CO2 evolved
from an unfumigated sample (control) often leads to low
or even negative biomass estimates because unequal
amounts of non-microbial biomass C is mineralised
(Horwath et al. 1996). To overcome this problem,
Jenkinson and Powlson (1976) suggested that CO2-C
released during the 10–20 day incubation rather than that
from the initial 0–10 day incubation of unfumigated soil
should be subtracted from the CO2-C released from the
fumigated soil. Horwath et al. (1996) suggested that the
proportion of CO2-C subtracted from the unfumigated
(0–10 day incubation) soil should vary as a function of
the ratio of CO2-C fumigated/CO2 control. When the
ratio is large the proportion of CO2-C subtracted from
the unfumigated soil should be large and vice versa.
They also suggested that equation (1) can be modified to:
Biomass C = (0.71 x CO2-C fumigated
– 0.23 x CO2-C control)/kC.
However, the modified equation needs to be validated
for soils under different land use and management and in
different climates.
The 2 basic assumptions mentioned above do not hold
for soils with pH <5, air-dried soils, waterlogged soils,
and soils that contain recently added organic materials or
plant residues. In acidic soils, the re-establishment of a C
and N mineralising microbial population after
fumigation and reinoculation is very slow. This causes a
 Microbial biomass: a review
reduced mineralisation of the killed microorganisms
which makes the usual k C and k N factors invalid
(Jenkinson 1988; Martens 1995). In air-dried soils, the
amount of already dead microorganisms may constitute
most of the microbial biomass in both fumigated and
unfumigated soil samples, in addition to the less
effective lysing of microbial cells by chloroform
(Sparling and West 1989). In waterlogged soils, CO2 and
CH4 are produced under conditions that restrict diffusion
of gases (Jenkinson 1988). In soils with recently added
organic materials or plant residues, the second
assumption is not met since the mass of the
re-established microbial population in the fumigated and
reinoculated soil sample corresponds to only 10–20% of
the original microbial biomass and consists mainly of
bacteria. This can be avoided by either careful removal
of the amendments such as roots, or a sufficient
preincubation of at least 3 weeks (Martens 1995).
Chloroform fumigation extraction
The above-mentioned limitations of the CFI method
are mainly overcome by extraction of C and N with
0.5 mol K2SO4/L from the chloroform fumigated and the
unfumigated soil samples. The proportions of C (kEC)
and N (k EN ) extracted from the fumigated (killed
microbial biomass) soil vary from 0.2 to 0.68 (Jenkinson
1988; Martens 1995). However, most frequently used
kEC values are in the range 0.36–0.45, while the kEN
values are in the range 0.49–0.62.
Likely limitations of the CFE method are differential
extraction of released C from soils that differ in clay
content and clay mineralogy, and variable k values
(Martens 1995).
The CFE method has been successfully used to
estimate soil microbial biomass P (Hedley and Stewart
1982) and S (Saggar et al. 1981). Inorganic P is
extracted with 0.5 mol Na2HCO3/L (pH 8.5) from both a
fumigated and an unfumigated soil; the proportion of P
is extracted from the killed microbial biomass, and the kP
value is taken as 0.4. The allowance is also made for P
sorption during fumigation and extraction by including
an internal P standard. For strong P retention soils such
as Ferrosols, Bray extractant (30 mmol NH 4 F/L +
25 mmol HCl/L) appears to be more appropriate than
0.5 mol Na2HCO3/L extractant (Oberson et al. 1997).
The procedure for microbial biomass S determination
is similar to that for microbial biomass P. The most
commonly used kS value is 0.41 (Smith and Paul 1990).
Substrate-induced respiration
An excess of substrate, usually glucose, is added to a
soil, which is then incubated at constant temperature and
moisture, and the respiration rate, CO2 evolved per hour,
is measured during a 0.5–2.5 h period, before the
microorganisms start proliferating and actually increase
microbial biomass.
651
Limitations of this method are: (i) that the pattern of
soil microbial response to glucose differs between soils;
(ii) that only glucose responsive soil microbial biomass
is measured; (iii) that soils recently amended with
organic materials or plant residues contain a large
proportion of young cells, and, therefore, the conversion
factor used, from mL CO2/h to microbial biomass C of
40 (30 at 22 o C, Beck et al. 1997) for an average
population in soil, is not valid (Martens 1995); (iv) it
measures only microbial activity which does not
necessarily equate with microbial biomass; and (v) that
microbial biomass N, P and S cannot be measured
(Smith and Paul 1990).
Adenosine triphosphate analysis
Adenosine triphosphate is a universal constituent of
living microbial cells. Although ATP can occur in dead
microbial cells and extracellularly in soil, it is rapidly
degraded by microorganisms. Therefore, ATP
concentration in soil can be used to estimate the amount
of living microbial biomass. It is usually extracted with
acid reagents from moist, preincubated soil and
estimated by the luciferin–luciferase system. The C : ATP
ratio is about 200 although it varies from 120 to 240
(Jenkinson et al. 1979; Eiland 1983; Martens 1995).
The limitations of the ATP method are: (i) that ATP is
decomposed by enzymatic and chemical hydrolysis
during the extraction process; (ii) after its release from
microbial cells, ATP is strongly sorbed by soil
constituents (Martens 1995); (iii) biomass C : ATP ratio
changes substantially over time in response to soil
amendments such as organic materials and plant residues
(Tsai et al. 1997); and (iv) it cannot measure microbial
biomass N, P and S in soil (Smith and Paul 1990).
Phospholipid fatty acids
Phospholipid fatty acids, with a chain length of <20 C
atoms are considered to be of mainly bacterial origin
(Harwood and Russel 1984). However, 18-C chain
phospholipid fatty acid, 18 : 2ϖ6 fatty acid, constitute on
average, 43% of the total phospholipid fatty acid in soil
fungi (Federle et al. 1986). Since ergosterol is specific to
the fungal membrane (Seitz et al. 1979), the fungal
biomass can be estimated from the correlation between
the amount of 18 : 2ϖ6 fatty acid and the ergosterol
content. Frostegard and Baath (1996) observed a close
correlation between the amounts of 18 : 2ϖ6 fatty acid
and the ergosterol in soil (r = 0.92), thus, indicating that
this phospholipid fatty acid can be used to estimate
fungal biomass. The ratio of 18 : 2ϖ6 fatty acid : bacterial
phospholipid fatty acid are then used as a
fungal : bacterial biomass ratio (Frostegard and Baath
1996). Phospholipid fatty acids can be extracted from
soil with a one-phase mixture of chloroform, methanol
and citric acid buffer, fractionated into neutral, glyco-
652 R. C. Dalal

and phospholipids on columns containing silicic acid, agricultural and forest soils in Table 1 show that all
methylated into fatty acid methyl esters and then 3 methods estimated similar soil microbial biomass in
measured on a gas chromatograph/mass spectrometer. the former but not in forest soils which had substantial
The advantage of the phospholipid fatty acid method, amounts of litter mixed with the soils. As expected, the
compared with other methods to estimate the microbial CFI method estimated the lowest amount of microbial
biomass of individual communities, is that both fungal biomass in the forest soils. Similar trends would be
and bacterial biomass can be estimated by the same expected in agricultural soils recently amended with
technique in a single soil extract (Frostegard and Baath organic materials or plant residues.
1996). The main disadvantages of the method are the However, unintentional variations in the experimental
instrumentation costs, incomplete and variable extraction procedure and sample handling such as sampling
of phospholipid fatty acids, possibility of including non- methods, sieving and partial removal of plant materials
living microbial biomass, and exclusion of other are not only a source of error in comparison between
microbial communities such as protozoa and nematodes. methods but also in the sequential estimations of
Comparison of different methods to estimate soil microbial biomass from the field. Thus, Joergensen
microbial biomass (1996) observed that the CFE-microbial biomass C : CFI-
All currently used soil microbial biomass methods microbial biomass C ratio varied more between
have some limitations since these were developed for laboratories than between different types of land use.
soils with microbial biomass in a relatively steady state. This should be kept in mind while comparing microbial
Furthermore, soil microbial biomass has been measured biomass values cited in the literature.
with variants of these methods, with different k factors,
soils at different moisture contents, different incubation Factors affecting the amount of soil microbial biomass
temperatures, soils containing variable amounts of Temperature and moisture
organic materials or plant residues, and different Besides organic substrates, temperature and moisture
instrumentation and analytical techniques. Therefore, it predominantly determine the amount of microbial
is difficult to compare soil microbial biomass values biomass in a soil (Wardle and Parkinson 1990). In a
obtained by different methods in different laboratories. geographically contiguous region, Dalal and Mayer
A comparative assessment of CFI, CFE and SIR (1987) found that microbial biomass increased with
methods was recently carried out by Beck et al. (1997) increasing mean annual precipitation; however, it
and Martens (1995). The results presented for decreased with mean annual temperature increase above
20oC in a semi-arid subtropical environment. Similarly,
Alvarez et al. (1995) observed that as the temperature
Table 1. Comparison of microbial biomass (mg C/kg soil) increased from 10 to 20oC microbial biomass declined
of arable and forest soils estimated by chloroform fumigation from 300 mg C/kg soil to 200 mg C/kg soil in an
incubation (CFI), chloroform fumigation extraction (CFE) and Argentinian Rolling Pampas soil. As temperature
substrate-induced respiration (SIR) methods increases the respiratory quotient (specific rate of
respiration per unit of microbial biomass C, usually
Soils CFI kC = 0.41 CFE kEC = 0.36 SIR F = 30C expressed as mg CO2-C respired/h or day.kg microbial
Arable soils biomass C) increases, which leads to C utilisation by
1A 222 235 251 surviving microbes from lysing microbial cells.
2A 442 323 413 Insam et al. (1989) observed that microbial biomass
3A 286 245 350 C as a proportion of total organic C (microbial quotient)
4B 299 276 296 decreased exponentially as the ratio of precipitation to
Mean ± s.d. 312 ± 93 270 ± 40 328 ± 70 pan evaporation (P/E) increased, that is,
Forest soils
1B 836 1129 744
Microbial biomass C/organic C (%)
2B 482 579 616 = 18.2 + 108.3 x exp(–7 P/E, mm) (3)
3B 1082 2094 1767 They hypothesised that soils exhibiting a microbial
4B 782 1746 1703 quotient higher or lower than predicted by equation (3)
5B 796 896 1154
would be accumulating or losing C respectively.
Mean ± s.d. 796 ± 213 1289 ± 621 1197 ± 530
Interestingly, Insam (1990) observed only a small effect
A Martens (1997). of pH, soil N or clay content on both microbial biomass
B Beck et al. (1997), with CFE values recalculated using kEC values and the microbial quotient. These observations need to
of 0.36 (Vance et al. 1987). be confirmed by others.
C SIR calculated from maximum initial respiration rate
Seasonal fluctuations in soil microbial biomass occur
(mL CO2/kg soil.h) x 30 (Beck et al. 1997).
due to changes in the amount of substrates, and
 Microbial biomass: a review 653

Table 2. Seasonal effects on microbial (mg/kg soil) carbon, Table 3. Effect of clay content on soil microbial biomass
nitrogen, phosphorus and sulfur in a sandy loam soil under pasture
Adapted from Sarathchandra et al. (1989) Clay Microbial biomass Biomass C : organic C
content (%) (mg/kg soil) (microbial quotient) (%)
Season Microbial biomass C:N:P:S
(night/day, oC) C N P S 6A 43 0.6
12A 83 0.6
Autumn (7/11) 839 140 80 10 84 : 14 : 8 : 1 12B 280 —
Winter (5/9) 664 163 80 12 55 : 14 : 7 : 1 13C 280 0.7
Early spring (11/15) 730 197 116 15 49 : 13 : 8 : 1 16A 262 1.4
Late spring (14/18) 1477 171 73 14 106 : 12 : 5 : 1 18D 213 1.7
20C 453 2.1
26C 460 2.8
34A 287 1.2
temperature and moisture. For example, Lynch and
34D 361 2.4
Panting (1982) found that the amount of microbial 40D 463 2.7
biomass reached a maximum around the time of 42B 750 —
maximum root biomass and thereafter declined. Under 43C 741 3.2
simulated temperature changes, Sarathchandra et al. 49D 508 2.3
(1989) observed higher microbial biomass values in 72D 453 2.8
autumn and late spring and lower values in winter A Sorensen (1983), agricultural soils.
although both microbial biomass N, P and S B Van Veen et al. (1985), protected microbial biomass.
accumulated during winter and early spring and declined C Powlson and Jenkinson (1981), zero tillage soils.
in late spring (Table 2). Also, the C : N : P : S ratio was D Dalal and Mayer (1987), virgin grassland and forest soils.
lower during winter and early spring (Table 2). They
speculated that this was a result of changes in the
microbial population. increasing clay content up to at least 50% of the clay
He et al. (1997) measured the seasonal responses in content of soil (Table 3).
microbial C, P and S in soils under pasture from March There is a paucity of microbial biomass data available
to December in Northumberland, England. From on soils containing more than 50% clay. It appears,
October to December, when plant residues were largely however, that in relatively undisturbed ecosystems, a
incorporated into the soils, biomass C and S increased plateau in microbial biomass values is reached in soils
1.5–2-fold. However, biomass P decreased during the around 50% clay content. It is possible, however, that the
dry summer, from July to September, and showed no fumigation efficiency of chloroform (CFI method) or
increase from October to December. Also, they did not extraction of metabolic products after fumigation (CFE
find any relationship between air temperature and method) may be lower in high clay content soils,
biomass C, P and S during the study period. especially those containing large numbers of fine
It is recognised that seasonal changes (temperature, micropores such as soils containing smectitic clays.
moisture and substrate supply) alter both the Badalucco et al. (1997) suggested that the lysing
composition and function of soil microbial biomass efficiency of chloroform could be influenced by its
(Zogg et al. 1997) but the trends in microbial C, N, P diffusion path through soil pores; fine micropores
and S are not well understood. A better understanding of provide a long and tortuous path and thus retard
such seasonal trends have implications in management diffusion of chloroform through these pores.
of organic matter turnover and nutrient cycling, and
hence regulation of N, P and S supply to plants.

Soil matrix Table 4. Microbial biomass carbon (mg/kg soil) and microbial
The soil matrix serves at least 3 functions: it provides quotient (%) in various size fractions of a silty loam soil
the capacity to preserve microbial biomass; it holds Adapted from Van Gestel et al. (1996)
substrates in the vicinity of microbial cells; and it
provides an environment for the efficient utilisation of Size fraction (mm) Microbial biomass C Microbial quotient
metabolic products for biosynthetic reactions. For >0.2 122.0 1.5
example, the amount of microbial biomass usually 0.2–0.05 283.0 1.8
increases with an increase in the clay content of a soil 0.05–0.02 63.9 2.3
(Powlson and Jenkinson 1981; Sorensen 1983; Van Veen 0.02–0.002 327.5 2.6
et al. 1984; Dalal and Mayer 1987). Microbial biomass <0.002 132.5 4.1
and the microbial quotient in soil increased with Whole soil 873.0 2.0
654
In aggregated silty loam soils, microbial biomass is
usually concentrated in the 0.020–0.002 mm size
fractions although microbial quotient increases as the
aggregate size is reduced (Table 4), the latter reflecting
at least 2 important functions of the clay fraction
mentioned earlier: a capacity to protect microbial
biomass, and a storage space for substrates near microbes.
Van Gestel et al. (1996) showed that the microbial
biomass increased from 122 mg C/kg soil in the >0.2 mm
fraction to 328 mg C/kg soil in the 0.020–0.002 mm
fraction while the <0.002 mm fraction contained only
132.5 mg C/kg soil. However, the microbial quotient
steadily increased from 1.5 to 4.1% as the fraction size
decreased from >0.200 to <0.002 mm in silty loam soil.
This phenomenon is incorporated into most of the
models of organic matter cycling in soil, which are based
on the assumption that the finer fraction of the soil
matrix, mostly the clay fraction, provides a protective
capacity to microbial biomass and organic matter (Van
Veen et al. 1985; Parton et al. 1987). Hassink and
Whitmore (1997) suggested that the physical capacity of
a soil to preserve organic matter and hence the maximum
amount of microbial biomass in a soil is limited. This is
presented in Figure 1 as a simplified adaptation of the
Hassink and Whitmore (1997) model of physical
protection of organic matter and microbial biomass in
soil. They suggested that the net rate of accumulation of
organic matter depends not on the protective capacity of
a soil but on the extent to which this capacity is already
occupied by organic matter. Similarly, the extent of the
increase in microbial biomass, for example, on the
addition of organic residues to a soil, should depend on
Residues
Figure 1. Turnover of organic matter through microbial biomass where protection is explicitly modelled by
sorption–desorption kinetics. NOM, non-protected organic matter; POM, protected organic matter; X, capacity
of soil to protect organic matter and microbial biomass; Ka, rate constant for protection/sorption; Kd, rate
constant for desorption (adapted from Hassink and Whitmore 1997).
R. C. Dalal
the soil’s capacity already occupied by microbial
biomass. This needs to be verified whether the potential
capacity of soil to protect and retain microbial biomass
varies not only with the finer fraction of the soil but also
climate (Insam et al. 1989; Insam 1990).
Land use
Microbial biomass usually declines when soils under
forest and grassland vegetation are brought under
cultivation (Srivastava and Singh 1991). For example,
Smith and Paul (1990) estimated 560, 680 and 870 mg
microbial biomass C/kg soil for cultivated, forest and
grassland systems respectively.
In southern Queensland, Dalal and Mayer (1987) found
that microbial biomass varied from 361 mg C/kg in belah
(Casuarina cristata) forest to 508 mg C/kg in brigalow
(Acacia harpophylla) forest soils. Soil microbial biomass
in these soils (Vertisols) when brought under cultivation
decreased at the rates of 4–14 mg C/kg soil.year (Table 5),
with an approximate turnover rate of 2.1 years. Basu and
Behera (1993) measured even higher losses in an ultisol
(31 mg C/kg soil . year) from rice cropping, which was
originally under forest (Shorea, Terminalia, Adina,
Anogeissus spp.).
Since turnover rates of soil microbial biomass vary
from 0.5 to 5 years compared with >20 years for the bulk
of soil organic matter, Powlson and Jenkinson (1981)
and Powlson et al. (1987) suggested that the changes in
microbial biomass could be used as an early predictor of
changes in soil organic matter due to various
management practices. In fact, Staben et al. (1997)
attempted to assess soil quality after wheat–fallow when
CO2
Microbial
NOM
biomass
Ka Kd
POM
X
 Microbial biomass: a review 655

Table 5. Rate of loss of microbial biomass from Vertisols subjected and Singh 1993). These effects on microbial biomass are
to cropping mainly due to the amount of C inputs through plant
Adapted from Dalal and Mayer (1987) growth and return of plant residues to the soil. For
example, Dalal et al. (1995) measured average C inputs
Soil type Clay Organic C Microbial Rate of loss
through root biomass and return of plant residues to be
content (%) biomass (mg C/kg . year)
(%) (mg C/kg soil)
1800, 2200 and 4800 kg C/ha . year from continuous
wheat cropping, medic–wheat rotations and grass +
Billa Billa loamy clay 34 1.58 361 6.3 legume pastures respectively. After 4 years, the
Cecilvale clay 40 1.65 464 7.5 respective microbial biomass C values were 426, 513
Langlands-logie clay 49 2.23 508 14.3 and 837 kg/ha (Table 6), thus, reflecting the extent of C
Waco clay 72 1.68 453 4.4 inputs to the soil.
However, in an alfisol, microbial biomass C
these soils were converted to a Conservation Reserve decreased due to the application of 80 kg N/ha.year for
Program. They found no significant differences in either 8 years to wheat, or including a legume in cereal
microbial biomass C or organic C and only C cropping, presumably due to lowering of soil pH (Ladd
mineralisation potentials increased in soils brought under et al. 1994). Carter (1986) also found that addition of
the Conservation Reserve Program. gypsum to an acidic soil reduced microbial biomass,
Obviously, more work should be undertaken before again by reducing soil pH, while addition of lime
microbial biomass could be used routinely as an increased microbial biomass. These effects could not
indicator of soil quality (refer to ‘Microbial biomass as entirely be ascribed to the amount of plant residues
an indicator of soil quality’), preferably by returned to the soil (Ladd et al. 1994) although root C
distinguishing 3 functions of microbial biomass: as a inputs were not measured.
nutrient source and sink, and as an agent governing the Application of organic amendments and farmyard
rates of C, N, P and S transformations. manure to soil increases microbial biomass. Ghosal and
Singh (1995) measured a 50% increase in microbial
Crop rotations, fertilisers and soil amendments biomass in a rice and lentil cropping system when
Crop rotations affect soil microbial biomass by farmyard manure was applied at the rate of 20 t/ha for
regulating the quantity and quality of plant biomass 4 years; application of 40 kg N/ha . year with farmyard
input, especially root biomass. The greater the input of manure increased microbial biomass from 200 to 350 mg
plant biomass, the more the increase in soil microbial C/kg soil. Similar increases were measured for microbial
biomass (Collins et al. 1992). For example, biomass N while microbial biomass P increased more
cereal–pasture rotation increases microbial biomass than 2-fold.
in soil (Table 6), especially in the top soil layers It is apparent, therefore, that rotations and additives
(Collins et al. 1992). which increase plant biomass production without a
Cereal cropping with N fertiliser applications also deleterious effect on soil pH usually lead to an increase
appears to increase microbial biomass C and N in soil in microbial biomass in soil; organic additives can
(Table 6; Campbell et al. 1991; Dalal et al. 1991; Singh directly increase microbial biomass unless these are
contaminated by heavy metals (Brookes and McGrath
Table 6. Effects of crop rotations and amendments on soil 1984) or adversely affect soil pH.
microbial biomass

Rotation Organic C Microbial Microbial Tillage practices

(%) biomass C biomass N Chemical control of weeds and absence of tillage for
(mg C/kg soil)(mg N/kg soil) seedbed preparation, that is, zero tillage practice, has led
to entirely new cropping and soil management practices.
Continuous wheatA 2.64 625 64 Soils that are zero-tilled develop a gradient of organic
Cont. wheat + 82 kg N/ha . yearA 2.79 686 83 matter which decreases more rapidly with depth than that
Wheat–wheat–sweet cloverA 2.63 658 83 in the conventionally tilled soils. The distribution of
Cont. wheatB 0.77 426 53 microbial biomass with depth in soil under zero tillage
Medic–wheatB 0.77 513 64
practice also follows a similar pattern to that of organic
Wheat–pasture (1.5 year)B 0.84 570 71
Wheat–pasture (2.5 year)B 0.91 806 100
matter (Doran 1980; Powlson and Jenkinson 1981;
Pasture (3.5 year)B 0.94 837 105 Doran 1987; Dalal et al. 1991). In fact, it has been
suggested that microbial biomass may provide an early
A Values calculated from Campbell et al. (1991) for 0–7.5 cm depth indication of changes in soil quality after introducing
after 11 years of rotations. zero tillage practice (Carter and Rennie 1982). For
B Dalal et al. (1994) for 0–5 cm depth after 4 years of treatments. example, Gupta et al. (1994) observed a significant
656 R. C. Dalal

increase in microbial biomass C and N in the top 5 cm of Table 7. Depth distribution of microbial biomass in soil under
an Alfisol only after 1 year of zero tillage practice, zero tillage (ZT) and conventional tillage (CT)
especially when crop residue was retained, while organic
Soil depth Biomass C (kg/ha) Biomass N (kg/ha)
C and total N were similar in both conventional tillage (cm) ZT CT ZT CT
and zero tillage practice. The microbial quotient
responded to the zero tillage practice, but not to crop Waco clayA
residue retention. Even after 7 years of zero tillage 0–2.5 167* 148 20.9* 18.6
practice, respective microbial biomass C and N in the 2.5–5 124 149* 15.5 18.6*
5–10 193 298* 24.1 37.3*
0–2.5 cm depth of soil were 35 and 30% higher than that
0–5 291 298 36.4 37.3
under conventional tillage (Haines and Uren 1990), 0–10 485 596* 60.6 74.5*
although no significant effect of tillage practice was
Melfort clay loamB
observed on organic C and total N in the soil. In the drier
0–5 389* 280 50* 36
environment of western New South Wales, however, 5–10 323 267 28 32*
microbial biomass, organic C and total N were not 0–10 712 547 78 68
significantly affected even after 13 years of zero tillage
Rothamsted silt loamC
practice (Fettell et al. 1994). This was presumably due to 0–20 997 1065 103 110
low crop yields and hence low amounts of crop residues
Boxworth clayC
returned to the soil.
0–20 1734* 1472 159 143
It has frequently been observed that, although zero
tillage practices result in greater concentration gradients A Calculated from Dalal et al. (1991), after 20 years of ZT practice.
with soil depth of organic C, total N, and microbial B From Carter and Rennie (1982), after 12 years of ZT practice.
biomass C and N than conventional tillage, the total C Adapted from Powlson and Jenkinson (1981), after 5–6 years of
amounts of these constituents may remain essentially ZT practice. * P<0.05.
similar in the top ≥10 cm depths under both tillage
practices (Powlson and Jenkinson 1981). Microbial
biomass C and N is usually greater in soil under zero Microbial biomass as a source of nutrients
tillage only in the top 0–2.5 or 0–5 cm depths but not Source of potentially mineralisable nitrogen
necessarily in the top 20 cm of soil (Table 7). Potentially mineralisable N is a measure of maximum
Thus, introduction of zero tillage practice does not N mineralised, under defined conditions, from soil
necessarily result in increased microbial biomass in the organic N that may become available to plants; the
soil profile, although the depth distribution of microbial magnitude of the amounts mineralised depends on
biomass may differ in soil between zero tillage and temperature, moisture, soil matrix and the N source. It
conventional tillage practices. has been suggested that the N source of potentially
Limitations of the microbial biomass values mineralisable N in soil is primarily the microbial
It appears that the soil microbial biomass values biomass N (Paul 1984; Adams and Attiwill 1986;
qualitatively express the effects of climate, land use and Myrold 1987).
management, and cultural practices but it is difficult to Potentially mineralisable N in soil is determined by 2
compare these values across climate, soil types and land biological methods; aerobic incubation and anaerobic
uses (Sparling 1997). incubation (Keeney 1982). The duration of aerobic

Table 8. Soil microbial biomass phosphorus and NaHCO3-extractable phosphorus (available P) in soil (adapted from Sparling et al. 1985)
Available phosphorus was measured on air-dried soils and values in parentheses are increases in available phosphorus on air drying (22oC)

Soil type Soil order pH Organic C (%) Biomass PA (mg/kg ) Available P (mg/kg)

Waikiwi (no P) Distrochrept 5.8 7.4 33.3 13.5 (6.7)

Waikiwi (20 kg P/ha.year) Distrochrept 5.8 4.5 15.5 17.4 (5.7)
Waikiwi (40 kg P/ha.year) Distrochrept 5.8 4.4 18.8 29.3 (6.8)
Lismore Ustrochrept 5.9 3.4 39.0 64.6 (14.2)
Molyneux Camborthid 5.9 1.7 9.8 13.9 (2.9)
Hokio Udipasamment 6.3 1.7 18.3 34.3 (–7.0)
Arapohue Lithic Rendoll 7.0 5.7 137.8 29.3 (15.2)

A Using kP value (proportion of P extracted from the killed microbial biomass) of 0.4 (Hedley and Stewart 1982).
 Microbial biomass: a review
incubation varies from 1 to 40 weeks (using either a
static or a leaching procedure), and that of anaerobic
incubation from 1 to 4 weeks, usually at 30 or 40oC.
Because of this variability in the procedures used, it is
difficult to equate the reported values of potentially
mineralisable N to that of microbial biomass in soil. In
many instances only associated relationships can be
established.
Adams and Attiwill (1986) hypothesised that most of
the NH 4 -N released during anaerobic incubation is
derived from mineralisation of N from dead aerobic
microorganisms killed by anaerobic conditions. Myrold
(1987) carried out anaerobic incubation (40oC, 7 days) in
soils containing 15N-labelled microbial biomass. He
found that the amount of NH4-N mineralised during
anaerobic incubation was closely correlated with that
mineralised in the CFI method. The equation describing
this was:
Anaerobic-N = 23 + 0.99 CFI-N (R2 = 0.81, P<0.01) (4)
Dalal et al. (1991) also obtained similar results on a fine-
textured soil subjected to different crop residue
management, fertiliser rates and tillage practices for
20 years. The equation was:
Anaerobic-N = –8.2 + 0.97 CFI-N (R2 = 0.52, P<0.01) (5)
However, it appears that not all N mineralised during
the anaerobic incubation is derived from microbial
biomass N (equations 5 and 6). Bonde et al. (1988)
found that only 55–89% of N released during anaerobic
incubation was derived from microbial biomass. This
was corroborated by Stockdale and Rees (1994). They
also observed that microbial biomass N constitutes a
variable, although significant, part of the potentially
mineralisable N in soil.
The amount of potentially mineralisable N measured
over 40 weeks of incubation far exceeds the amount of
microbial biomass N (6–10% v. 2–6% of total N) in soil
(Dalal and Mayer 1987; Bonde et al. 1988). Apparently,
a substantial amount of non-biomass N but labile N is
also mineralised during the long period of incubation.
Rice et al. (1996) concluded that the relationship
between microbial biomass and mineralisable N can be
influenced by N fertilisation history and the contribution
of non-living or dormant microbial biomass to N
mineralisation. Bonde et al. (1988) estimated that
67–78% of total microbial biomass consist of a dormant
inactive fraction, which could be potentially mineralised
during the incubation period. Potentially mineralisable N
also increases with the addition of N through fertiliser,
plant material or farmyard manure, which enters the
labile fraction of soil organic N (Bonde et al. 1988).
The implication is that the role of microbial biomass
N lies not only in supplying N from its biomass during
plant growth, especially after dry–wet cycles, but also to
657
transform non-biomass labile N into a mineral form for
the plant needs.
However, it is uncertain whether the microbial
biomass provides potentially mineralisable N through its
reduction in size or by mineralising labile organic N
fraction during a growing season. Turnover rates of
microbial biomass range from 50 to 2000 days (Smith
and Paul 1990) while gross N transformation rates
by and through microbial biomass are an order of
magnitude faster (Shen et al. 1984).
Measuring these pools as well as their rates of
transformation is critical in accounting for the plant-
available N pool in soil. This is further complicated by
the fact that during rapid plant growth, the size of the
microbial biomass may even increase or remain similar
due to C supply from root exudates (Dinwoodie and
Juma 1988), and thus it may compete with plants for N,
albeit for a small amount (Robertson et al. 1997).
Fortunately, for most arable crops, the competitive
demand for N by microbial biomass is small except
when fresh plant residues have recently been added to
soil.
Microbial biomass contribution to available phosphorus
and sulfur
The potential contribution of microbial biomass P to
plants may be assessed from the increase in available P
in soil when the soil is dried, presumably from killed
microbial biomass. For example, Sparling et al. (1985)
showed a good agreement in pasture soils between the
increase in available P and the size of microbial biomass
P when the soils were air-dried (Table 8). The
contribution of microbial biomass P to available P varied
from 4 to 76%. It was greater in soils containing greater
amounts of organic matter (>2% organic C). This
phenomenon of varying amounts of microbial biomass P
mineralised during air-drying of soils has implications
for soil testing for available P in the laboratory. Since it
is not practical to use routinely moist soil samples for the
determination of available P, a standard procedure must
be used in air-drying soils for interpretations of available
P levels across regions and soils.
However, it is not certain whether microbial biomass
P released during drying of the soil may still be available
to plants when the soil is wetted. In fact, McLaughlin
et al. (1988a) observed that the amounts of microbial
biomass P in soil increased linearly with increasing soil
water content in the field, with a large and rapid increase in
the amount of P incorporated into the microbial biomass in
the first 7 days after wetting the initially dry soil.
McLaughlin et al. (1988b) also found that the amount
of microbial biomass P (27.9 kg P/ha) in a solonetz soil
was almost 4 times the amount that was taken up by the
wheat crop (7.5 kg P/ha) in 95 days. However, there is a
scarcity of information on microbial biomass P turnover
658
Table 9. Microbial biomass phosphorus, microbial biomass sulfur,
available phosphorus and available sulfur in a clay loam pasture
soil under different treatments (from He et al. 1997)
Treatments since 1897 include N applied at 35 kg N/ha.year
(+ 19 kg S/ha.year), P at 26 kg P/ha.year (+ 2 kg S/ha.year),
NPK (nitrogen, phosphorus, potassium) include N and P at the above
rates and 46 kg K/ha.year, FYM (farmyard manure) at 20 t/ha.year, and
FYM + NPK applied at 20 t FYM/ha.year and NPK at half the straight
NPK rates
Biomass P and available P were extracted by 0.03 mol NH4F/L +
0–025 mol HC/L, and kP value (proportion of P extracted from killed
microbial biomass) of 0.4
Biomass S and available S were extracted by 0.01 mol CaCl2/L, and kS
value (proportion of S extracted from killed microbial biomass) of 0.31
(Wu et al. 1994)
Treatment Biomass P Available P Biomass S Available S
(mg/kg soil) (mg/kg soil) (mg/kg soil) (mg/kg soil)
Control 37 0.4 19 2.8
N 11 0.7 17 15.5
P 154 10.6 24 5.1
NPK 88 15.5 25 6.9
FYM 193 129 27 5.7
FYM + NPK 178 172 18 6.3
and management practices that can influence both the
size and rate of turnover of microbial biomass P in soil
(Smith and Paul 1990; Horwath and Paul 1994).
In pasture soils, biomass P and biomass S constitute a
substantial source of available P and S, especially where
soil fertility has been enhanced by regular applications of
fertilisers and organic manures (Table 9). He et al.
(1997) found that the amount of P in the pasture herbage
was more closely related to biomass P (R2 = 0.83) than
to the available P (R2 = 0.49) present in the soil, although
the most significant correlation was found between the
amount of P in the herbage and the sum of both biomass
P and available P in the soil (R2 = 0.98).
Microbial biomass S as well as available S, however,
were poorly correlated with the herbage S yield,
presumably because 0.01 mol CaCl 2 /L was a poor
extractant of S available over the whole season of
herbage growth (He et al. 1997).
It should be emphasised that studies on the relative
contribution of P and S from microbial biomass to the
available pool and then to the plant are complicated by
simultaneous chemical and physical processes such as
sorption, diffusion into aggregates, occlusion and
precipitation operating on P and S mineralised from the
microbial biomass, and P uptake by the plant.
Limited information is available on rates of
transformation of microbial biomass of P and S. Smith
and Paul (1990) summarised the rates of N, P and S
mineralisation: the respective average rates of
mineralisation were 0.5 mg N/kg soil.day, 0.02 mg P/kg
R. C. Dalal
soil.day and 0.04 mg S/kg soil.day for arable soils while
for grassland soils the values were 2-fold higher.
However, these rates would include both the microbial
biomass as well as non-microbial N, P and S sources in
soil.
The average rate of microbial biomass C
mineralisation is usually taken as 2 x 10–3/day (Smith
and Paul 1990); a value similar to the maintenance
energy requirement for dormant organisms
(2–4 x 10–3/day, Anderson and Domsch 1985). However,
the individual N, P and S mineralisation rates of
microbial biomass vary with the C : N, C : P, C : S ratios,
moisture, temperature, pH and soil matrix (Smith and
Paul 1990).
Limitations of microbial biomass values as the
indicators of nutrient source
Although the amount of microbial biomass in a soil is
a potential source of nutrients to a crop, the
interpretation of its actual contribution to the crop needs
is complicated by the different microbial biomass
measurement techniques used, and the variables such as
moisture, temperature, pH, soil matrix, and
carbon : nutrient ratios that simultaneously affect
mineralisation–immobilisation processes, and
competitive nutrient demands. Moreover, P and S
sources and availability are also controlled by abiotic
factors (Smith and Paul 1990).
Microbial biomass as a sink of nutrients
Grasslands, woodlands and forests contain substantial
amounts of microbial biomass. For example, the average
microbial biomass pool size is 20–50% higher in
grasslands and forest soils than in arable soils (Table 1),
mainly because of the larger organic C inputs in the
former. Addition of easily metabolisable C to soil rapidly
increases microbial biomass, often by several fold,
and also incorporates nutrients into microbial
biomass rapidly, often within a week of addition
(Robertson et al. 1997; McLaughlin et al. 1988a).
However, such a microbial pool size is transient, and
the level of microbial biomass a soil can sustain under a
steady-state condition depends on the protective capacity
of the soil, which is governed by a dynamic interaction
between clay (Sorensen 1983; Van Veen et al. 1985), and
clay aggregates (Van Gestel et al. 1996), soil microbial
biomass and soil organic matter (Hassink and Whitmore
1997), and possibly climate (Insam and Haselwandter
1989; Insam 1990).
Different management practices also affect the levels
of microbial biomass in a soil (Table 6). For example,
Schnurer et al. (1985) reported more than a 100%
increase in microbial biomass C and N by the addition of
crop residues, and N fertiliser and manure applications
 Microbial biomass: a review 659

Table 10. Effects of zinc and copper contained in sewage sludge applied to a sandy loam soil on microbial biomass and microbial quotient
(adapted from Chander and Brookes 1993)
Sewage sludge was applied in 1982; the sludge was spiked with Zn and Cu singly or in combination to achieve variable Zn and Cu concentrations;
these concentrations were further adjusted with fresh applications of the Zn and/or Cu contaminated sewage sludge in 1986 (rates shown in
parentheses); crops grown since 1986 were barley until 1988 then clover ley for the next two years; the soil samples (0–10 cm depth)
were taken in 1990

Treatment Rate (t/ha) CaCl2 extractable (mg/kg) Microbial biomass Microbial quotient
Zn Cu (mg C/kg ) (%)

Control soil 0 42 12 169 1.63

Clean sludge 200 66 19 183 1.50
Zn sludge 200 220 29 185 1.50
Zn sludge 200 (82) 457 28 140 0.98
Cu sludge 200 (83) 74 197 150 1.06
Cu sludge 200 (139) 104 415 94 0.52
Zn–Cu sludge 200 (110) 367 191 120 0.66
Zn–Cu sludge 200 (162) 427 262 79 0.42

for 27 years to a clay loam soil. Similarly, microbial
biomass P values increased by 2–5-fold with the
application of P fertilisers and manures for over 90 years
(Table 9); however, these observations need to be
confirmed in different environments.
Limitations of microbial biomass values indicating the
nutrient sink source
Carbon flux through microbial biomass is the driving
force for the decomposition of plant residues and other
organic materials. While microbial biomass is building
up it can immobilise substantial amounts of N, P and S.
However, eventually C becomes limiting, microbial
biomass dies, and a large proportion of N, P and S is
transferred to the non-living microbial biomass. Thus,
microbial biomass values, at the most, indicate only the
partial source of nutrients for a crop. Moreover,
limitations concerning the variability in procedures of
measuring microbial biomass, and the variables affecting
it still apply.
Effects of heavy metals and pesticides
Increasing disposal of sewage sludge on land has the
potential to increase heavy metals in soil. There is thus
concern about the long-term effects of additions of heavy
metals in sewage sludge to soil. It is believed that
microbial biomass, being the living component of soil,
can provide an early indication of the adverse effects
of increasing concentrations of heavy metals in
soil (Brookes and McGrath 1984; Chander and
Brookes 1993; Brookes 1995; Valsecchi et al. 1995;
Dahlin et al. 1997).
Brookes and McGrath (1984) found that the amounts
of microbial biomass in agricultural soils treated with
either sewage sludge or sludge-containing composts
(average microbial biomass, 93 mg C/kg soil) were
much smaller than in soils which received farmyard
manure (205 mg C/kg soil) over the same period. As the
EDTA-extractable nickel and copper (Cu) increased,
microbial biomass decreased, and these effects were
detectable even after 20 years of application.
Furthermore, Chander and Brookes (1993) found that a
combination of zinc (Zn) and Cu at high concentrations
had an additive adverse effect on the amount of
microbial biomass present although, at similar
concentrations (CaCl2-extractable Cu and Zn, 415 and
457 mg/kg respectively), microbial biomass decreased
by 50% more with Cu than Zn (Table 10).
Also, the microbial quotient decreases much faster
than the total microbial biomass as the heavy metal
concentration increases. For example, the microbial
quotient decreased from 1.5% in soil that received
uncontaminated sewage sludge to 1% with Zn
(457 mg/kg soil) and 0.5% with Cu (415 mg/kg soil)
contained in the sludge applied (Table 10). Valsecchi
et al. (1995) found that large additions of heavy metals
through long-term effluent decreased the microbial
quotient more than an order of magnitude, from 4% in
the slightly contaminated soil to 0.2% in heavily
contaminated soil; the ratio of specific respiratory rate to
organic C decreased only by 50% (Table 11). On the
other hand, both the specific respiration rate and
respiratory quotient were much higher in the latter.
Thus, the microbial quotient provides a sensitive
indicator of the effects of heavy metals on soil microbial
biomass. This may be because either the soil microbial
population becomes smaller (Brookes and McGrath
1984) or soil microbial biomass becomes less effective
in mineralising soil organic matter, that is, turnover rate
of organic matter decreases (Valsecchi et al. 1995) as
organic matter accumulates in soil with the applications
of the heavy metals contaminated sewage sludge.
The higher respiratory quotient, combined with the
660
Table 11. Effect of long-term application of heavy metal polluted sewage effluent to three sandy loam soils on organic carbon, microbial
biomass carbon (MBC), specific respiration rate (SRR), microbial quotient (MBC/organic C), respiratory quotient (SRR/MBC), and
specific respiration rate/organic carbon(SRR/organic C) (adapted from Valsecchi et al. 1995)
Soil Ammonium acetate–EDTA extractable (mg/kg) Organic C
Zn Cu Pb Ni Cr Cd (%)
1 17.9 12.2 9.2 1.6 2.7 0.6 0.90
2 16.9 16.2 16.7 2.5 3.7 0.7 1.95
3 4884.0 228.0 771.0 24.0 30.0 41.0 5.60
lower ratio of specific respiratory rate to organic C, in
the heavy metals contaminated soil indicates a state of
microbial stress due to heavy metals toxicity. These
metals reduce the energy efficiency of the microbial
metabolic processes, which then require greater amounts
of C for maintenance, thus reducing the quantity of C
incorporated into the microbial biomass (Valsecchi et al.
1995).
Since the toxic effects of heavy metals on microbial
biomass are additive, it has been suggested that even
smaller concentrations of a combination of heavy metals
in sewage sludge may affect not only microbial biomass
and activity but also community structure as determined
by phospholipid fatty acids analysis (Dahlin et al. 1997).
This is expected since heavy metals such as Cu
adversely affect the fungal community. However, limited
information is available on the effects of heavy metals
on nutrient cycling by and through microbial biomass
although N and P hydrolysing enzymes have been found
to be adversely affected (Kuperman and Carreiro 1997).
A substantial proportion of cropping land receives
regular applications of herbicides and insecticides. While
herbicides applied at the field rate are decomposed in
soil with only a slight effect on the amount of microbial
biomass (Perucci and Scarponi 1994; Voos and
Groffman 1997), some insecticides applied to soil not
only reduce the size of microbial biomass, but also alter
the composition of the microbial communities (Spain
and van Veld 1983; Bromilow et al. 1996).
Perucci and Scarponi (1994) found that the herbicide,
imazethapyr (amidazolinone derivative) used to control a
wide spectrum of broadleaf weeds and grasses, when
applied at recommended field rate (50 g a.i./ha) had no
effect on soil microbial biomass although at 100-fold the
field rate it had an adverse effect on the amount of
microbial biomass. Similar results were obtained by
Voos and Groffman (1997) for the microbial breakdown
of 2,4-D and dicamba. They also observed that the rate
of dissipation of the herbicides increased with the
amount of microbial biomass in the soil. Bromilow et al.
(1996) also measured no adverse effects on the amount
of microbial biomass of annual application of aldicarb,
R. C. Dalal
MBC SRR MBC/organic C SRR/MBC SRR/organic C
(mg C/kg) (mg CO2-C/kg.day) (%) (%/day) (%/day)
366 3.66 4.06 1.01 0.04
555 8.99 2.85 1.62 0.05
125 11.48 0.22 9.18 0.02
benomyl, chlorfenvinphos, glyphosate, chlorotoluron or
triadimefon to soil, used for barley cropping, for up to
20 years.
The application of fungicides shifts the balance of
microbial biomass towards bacteria-dominated microbial
biomass (Spain and van Veld 1983). We need to know a
great deal about the effects of herbicides and pesticides,
and other organic additives, especially under repeated
applications on the 3 functions of microbial biomass in
soil: as a source and a sink of nutrients, and an agent of
transformation of nutrients.
Limitations of microbial biomass values as indicators of
heavy metal toxicity
Microbial biomass values as such do not usually
provide good indicators of heavy metal toxicity in soil.
Some enzyme activities (Kuperman and Carreiro 1997)
and susceptible plants are much more sensitive
indicators. However, some microbial indices such as the
microbial quotient or the ratio of specific respiratory
rate/organic C may prove useful.
Microbial biomass as an indicator of soil quality
Soil quality is the capacity of a specific kind of soil to
function, within natural or managed ecosystem
boundaries, to sustain plant and animal productivity,
maintain or enhance water and air quality, and support
human health and habitation (Karlen et al. 1997). This
definition should be extended to include ‘maintain or
enhance soil biodiversity’. To evaluate soil quality, the
capacity of the soil to function needs to be measured
using appropriate indicators. The most desirable
attributes of an appropriate indicator include the
following: (i) it measures one or more soil functions;
(ii) it is sensitive enough to measure changes due to
disturbance, restoration, or change in land use and
management; (iii) it provides benchmark, critical or
threshold values; (iv) it can be readily interpreted; and
(v) it is cost-effective.
Microbial biomass performs 3 essential functions in
soil (see ‘Introduction’). It is also sensitive enough to
measure the changes due to the different land use and
management such as virgin land brought under
 Microbial biomass: a review
cultivation (Dalal and Mayer 1987), crop rotations
(McGill et al. 1986), restoration (Staben et al. 1997) and
heavy metal contamination (Brookes and McGrath
1984). However, currently, microbial biomass does not
provide benchmark, critical or threshold values against
which soil quality can be evaluated.
The benchmark values will differ depending on soil
type and land uses (Sparling 1997), climate (Insam et al.
1989), and vegetation (Martens 1995). Such differences
can be reduced by comparing the derived microbial
indices such as the microbial quotient for soils of
different organic matter contents (Powlson and
Jenkinson 1981; Anderson and Domsch 1989; Sparling
1992) and respiratory quotient for soils of different
microbial biomass contents (Insam and Haselwandter
1989; Anderson and Domsch 1990). In addition, the
ratio of microbial-specific respiration rate/organic C has
been proposed as a sensitive indicator for heavy metal
contaminated soils (Valsecchi et al. 1995).
The benchmark values of the microbial biomass and
even the derived microbial indices for soil quality are not
available (Sparling 1997) although the differences in
both microbial biomass and microbial indices can be
demonstrated for different soil types and land uses and
management (Tables 1, 2, 5–11). Do such values exist?
Are these values available in the literature on soil
microbial biomass? With our current understanding of
soil microbial biomass and soil quality, the answer is not
in the affirmative. It is tempting to suggest that the
(maximum) potential amount of microbial biomass in a
soil could be estimated from the protective capacity of a
soil (Hassink and Whitmore 1997) for the specified
climate. By corollary with organic matter, the potential
microbial quotient can then be benchmarked.
However, the problem of interpretation of the
benchmark values and deviation from these values in
terms of soil quality is still unresolved. It is tempting to
suggest that the extreme stress on the soil’s ecosystem
can be identified from a combination of the derived
microbial indices (Table 11). From Tables 10 and 11, it is
suggested that the microbial quotient value of <0.5 could
be considered an indicator of the soil under stress (poor
soil quality, that is, the impairment of the soil’s capacity
to function). Further work is required to validate this
value for soil microbial biomass under stress due to
factors other than heavy metal contamination.
On the other hand, the significance of specific
respiratory rate and respiratory quotient on soil quality is
less clear. For example, both highly productive soils as
well as contaminated soils exhibit high respiratory rates
and high respiratory quotients (Brookes and McGrath
1984; Alvarez et al. 1995; Valsecchi et al. 1995;
Sparling 1997) although the ratio of specific respiration
rate to organic C should be higher in the former. How do
these measurements relate to the soil quality? The
661
common aspect of these 2 systems appears to be the low
incorporation of C into microbial biomass, and by
inference, probably leading to low soil microbial
diversity.
The ratio of fungal biomass to bacterial biomass
appears to be sensitive to land use changes. For example,
this ratio was found to be higher in a restorative soil
system than in the cultivated soil (Staben et al. 1997).
This was a reflection of the dominance of the bacteria-
feeding nematodes in the former; which had a ratio of
fungal-feeding nematodes to bacterial-feeding
nematodes of 0.5 compared with 4.8 for the cultivated
soil. However, the applicability of microbial diversity to
soil quality requires further study although its cost-
effectiveness may limit its measurement as a routine
measure of soil quality.
Limitations
Besides the current limitations of unavailability of
benchmark values and difficulty in interpretation, soil
microbial biomass measurements need to be cost-
effective. Moreover, current methods of measuring
microbial biomass, its activity, and biodiversity in soil
are laborious, cumbersome and suffer from a lack of
close association with productivity. Although microbial
measurements are essential in evaluating soil quality,
these limitations need to be overcome if soil microbial
biomass is to be routinely measured and used as an
indicator of soil quality.
Future perspectives
Smith and Paul (1990) asked the question, ‘What do
our microbial biomass values represent and how can we
use them to study nutrient cycling?’
Since 1990, microbial biomass measurements have
been extensively used to study nutrient cycling. We
know that although microbial biomass is only a small
component of soil organic matter, usually <5%, nutrient
fluxes through microbial biomass are at least an order of
magnitude faster than the remaining organic matter;
hence, its study is critical in understanding the nutrient
cycling in soil.
Due to the rapid turnover rate of microbial biomass, it
responds to changes in soil and crop management
practices much faster and earlier than the bulk of soil
organic matter. Therefore, microbial biomass
measurements have been critical in monitoring early
changes in soil induced by zero tillage practice,
restorative practices, use of herbicides and insecticides,
and application of sewage sludge to land.
For a given soil and cultural practice, soil microbial
biomass has been useful as an indicator of changes in
soil due to the above-mentioned practices. Therefore, it
has been suggested as an important biological indicator
of soil health (Pankhurst 1994; Sparling 1997) and soil
quality (Karlen et al. 1997). However, as a management
662
tool its applications have been limited because
analytically soil microbial biomass needs to have
measurements of all of its 3 functions simultaneously:
source size, sink size, and turnover rate for C, N, P and
S. As pointed out by Jenkinson (1988) while discussing
the methods of microbial biomass measurements in soil,
that more than one method (CFI, CFE, SIR, ATP
measurement) is required to properly estimate even the
size of the microbial biomass. This statement is
applicable even today if we are to properly evaluate the
various functions of soil microbial biomass and as an
indicator of soil quality. However, the time consuming
and cumbersome microbial biomass measurements
restricts its utility as a routine analysis in soil testing
laboratories.
Another major limitation of acceptance of soil
microbial biomass measurements still remains. What do
microbial biomass numbers really mean? There is only a
tenuous relationship between the size of microbial
biomass and crop yield. Does a microbial biomass–crop
yield response curve exist? Surrogatively, probably
(Insam et al. 1991). There is some indication that the
relationship between the size of microbial biomass and
respiration activity could differentiate between sluggish
and stressed states of soil health (Sparling 1997).
However, comparative ‘benchmark’ references of
microbial biomass as critical or threshold and optimum
levels do not currently exist. Will they ever exist? Only
further work will ascertain that.
Microbial biomass is central to organic matter
cycling, and hence, carbon sequestration by soil. What is
the capacity of soil to protect microbial biomass and how
does it relate to the optimum organic matter content of
soil? Data presented in this review provided only partial,
if any, answers to these questions.
Acknowledgments
I thank Ms C. McDowall and Ms M. Halliday for
assistance with the literature search, and anonymous
reviewers for numerous comments and suggestions.
References
Adams, M. A., and Attiwill, P. M. (1986). Nutrient cycling and
nitrogen mineralisation in eucalypt forests of south-eastern
Australia. II. Indices of nitrogen mineralisation. Plant and
Soil 92, 341–63.
Alvarez, R., Santanatoglia, O. J., and Garcia, R. (1995). Effect
of temperature on soil microbial biomass and its metabolic
quotient in situ under different tillage systems. Biology
and Fertility of Soils 19, 227–30.
Anderson, J. P. E., and Domsch, K. H. (1978). A
physiological method for the quantitative measurement of
microbial biomass in soils. Soil Biology and
Biochemistry 10, 215–21.
Anderson, J. P. E., and Domsch, K. H. (1985). Determination
of ecophysiological maintenance C requirements of soil
microorganisms in a dormant state. Biology and Fertility
of Soils 1, 81–9.
R. C. Dalal
Anderson, T. H., and Domsch, K. H. (1989). Ratios of
microbial biomass carbon to total carbon in arable soils.
Soil Biology and Biochemistry 21, 471–9.
Anderson, T. H., and Domsch, K. H. (1990). Application of
eco-physiological quotients (qCO2 and qD) on microbial
biomasses from soils of different cropping histories. Soil
Biology and Biochemistry 22, 251–5.
Angers, D. A., Pesant, A., and Vigneux, J. (1992). Early
cropping induced changes in soil aggregation, organic
matter, and microbial biomass. Soil Science Society of
America Journal 56, 115–19.
Atlas, R. M., and Bartha, R. (1981). ‘Microbial Ecology:
Fundamentals and Applications.’ (Addison-Wesley: Menlo
Park, CA, USA.)
Badalucco, L., De Cesare, F., Grego, S., Landi, L., and
Nannipieri, P. (1997). Do physical properties of soil affect
chloroform efficiency in lysing microbial biomass? Soil
Biology and Biochemistry 29, 1135–42.
Basu, S., and Behera, N. (1993). The effect of tropical forest
conversion on soil microbial biomass. Biology and
Fertility of Soils 16, 302–4.
Beck, T., Joergensen, G., Kandeler, E., Makeschin, E., Nuss, H.,
Oberholzer, R., and Scheu, S. (1997). An inter-laboratory
comparison of ten different ways of measuring soil
microbial biomass C. Soil Biology and Biochemistry 29,
1023–32.
Bonde, T. A., Schnurer, J., and Rosswall, T. (1988). Microbial
biomass as a fraction of potentially mineralizable nitrogen
in soils from long-term field experiments. Soil Biology and
Biochemistry 20, 447–52.
Bromilow, R. H., Evans, A. A., Nicholls, P. H., Todd, A. D.,
and Briggs, G. G. (1996). The effects on soil fertility of
repeated applications of pesticides over 20 years. Pesticide
Science 48, 63–72.
Brookes, P. C. (1995). The use of microbial parameters in
monitoring soil pollution by heavy metals. Biology and
Fertility of Soils 19, 269–70.
Brookes, P. C., and McGrath, S. P. (1984). Effects of metal
toxicity on the size of the soil microbial biomass. Journal
of Soil Science 35, 341–6.
Brookes, P. C., Powlson, D. S., and Jenkinson, D. S. (1985).
Phosphorus in the soil microbial biomass. Soil Biology and
Biochemistry 16, 169–75.
Campbell, C. A., Biederbeck, V. O., Zentner, R. P., and
Lafond, G. P. (1991). Effects of crop rotations and cultural
practices on soil organic matter, microbial biomass and
respiration in a thin Black Chernozem. Canadian Journal
of Soil Science 71, 363–76.
Carter, M. R. (1986). Microbial biomass and mineralisable
nitrogen in solonetzic soils: influence of gypsum and lime
amendments. Soil Biology and Biochemistry 18, 531–7.
Carter, M. R., and Rennie, D. A. (1982). Changes in soil
quality under zero tillage farming systems: distribution of
microbial biomass and mineralisable C and N potentials.
Canadian Journal of Soil Science 62, 587–97.
Chander, K., and Brookes, P. C. (1993). Residual effects of zinc,
copper and nickel in sewage sludge on microbial biomass in
a sandy loam. Soil Biology and Biochemistry 25, 1231–9.
Collins, H. P., Rasmussen, P. E., and Douglas, C. L. Jr (1992).
Crop rotation and residue management effects on soil
carbon and microbial dynamics. Soil Science Society of
America Journal 56, 783–8.
 Microbial biomass: a review
Dahlin, S., Witter, E., Martensson, A., Turner, A., and Baath, E.
(1997). Where’s the limit? Changes in the microbiological
properties of agricultural soils at low levels of metal
contamination. Soil Biology and Biochemistry 29, 1405–15.
Dalal, R. C., Henderson, P. A., and Glasby, J. M. (1991).
Organic matter and microbial biomass in a vertisol after
20 years of zero tillage. Soil Biology and Biochemistry 23,
435–41.
Dalal, R. C., and Mayer, R. J. (1987). Long-term trends in
fertility of soils under continuous cultivation and cereal
cropping in Southern Queensland. VII. Dynamics of
nitrogen mineralisation potentials and microbial biomass.
Australian Journal of Soil Research 25, 461–72.
Dalal, R. C., Strong, W. M., Weston, E. J., Cahill, M. J.,
Cooper, J. E., Lehane, K. J., King, A. J., and Gaffeney, J.
(1994). Evaluation of forage and grain legumes, no-tillage
and fertilisers to restore fertility degraded soils.
Transactions, XV International Society of Soil Science,
Acapulco, Mexico 5a, 62–74.
Dalal, R. C., Strong, W. M., Weston, E. J., Cooper, J. E., Lehane,
K. J., King, A. J., and Chicken, C. J. (1995). Sustaining
productivity of a vertisol at Warra, Queensland, with
fertilisers, no-tillage or legumes. 1. Organic matter status.
Australian Journal of Experimental Agriculture 35, 903–13.
Dinwoodie, G. D., and Juma, N. G. (1988). Allocation and
microbial utilization of C in two soils cropped to barley.
Canadian Journal of Soil Science 68, 495–505.
Doran, J. W. (1980). Soil microbial and biochemical changes
associated with reduced tillage. Soil Science Society of
America Journal 53, 1511–15.
Doran, J. W. (1987). Microbial biomass and mineralisable
nitrogen distribution in no-tilled and ploughed soils.
Biology and Fertility of Soils 5, 68–75.
Eiland, F. (1983). A simple method for quantitative
determination of ATP in soil. Soil Biology and
Biochemistry 15, 665–70.
Federle, T., Livingstone, R. J., Wolfe, L. E., and White, D. C.
(1986). A quantitative comparison of microbial community
structure of estuarine sediments from microcosms and the
field. Canadian Journal of Microbiology 32, 319–25.
Fettell, N. A., Carpenter, D. J., Kidston, J., and Gupta, V. V. S. R.
(1994). Organic matter, microbial biomass and respiration
after 13 years of stubble retention and zero tillage in
western NSW. In ‘Soil Biota: Management in Sustainable
Farming Systems’. (Ed. C. E. Pankhurst.) p. 177. (CSIRO:
East Melbourne.)
Frostegard, A., and Baath, E. (1996). The use of phospholipid
fatty acid analysis to estimate bacterial and fungal biomass
in soil. Biology and Fertility of Soils 22, 59–65.
Ghosal, N., and Singh, K. P. (1995). Effects of farmyard
manure and inorganic fertiliser on the dynamics of soil
microbial biomass in a tropical dryland agroecosystem.
Biology and Fertility of Soils 19, 231–8.
Gupta, V. V. S. R., Roper, M. M., Kirkegaard, J. A., and
Angus, J. F. (1994). Changes in microbial biomass and
organic matter levels during the first year of modified
tillage and stubble management practices on a red earth.
Australian Journal of Soil Research 32, 1339–54.
Haines, P. J., and Uren, N. C. (1990). Effects of conservation
tillage farming on soil microbial biomass, organic matter
and earthworm populations in north-eastern Victoria.
Australian Journal of Experimental Agriculture 30, 365–71.
663
Harwood, J. L., and Russel, N. J. (1984). ‘Lipids in Plants and
Microbes.’ (George Allen and Unwin: London.)
Hassink, J., and Whitmore, A. P. (1997). A model of the
physical protection of organic matter in soils. Soil Science
Society of America Journal 61, 131–9.
He, Z. L., Wu, J., O’Donnell, A. G., and Syers, J. K. (1997).
Seasonal responses in microbial biomass carbon,
phosphorus and sulphur in soils under pasture. Biology and
Fertility of Soils 24, 421–8.
Hedley, M. J., and Stewart, J. W. B. (1982). Method to measure
microbial biomass phosphorus in soils. Soil Biology and
Biochemistry 14, 377–85.
Horwath, W. R., and Paul, E. A. (1994). Microbial biomass.
In ‘Methods of Soil Analysis’. Part 2. Microbiological and
Biochemical Properties. (Eds R. W. Weaver et al.)
pp. 754–73. (Soil Science Society of America: Madison,
Wisconsin, USA.)
Horwath, W. R., Paul, E. A., Harris, D., Norton, J., Jagger, L.,
and Horton, K. A. (1996). Defining a realistic control for
the chloroform fumigation-incubation method using
microscopic counting and 14 C-substrates. Canadian
Journal of Soil Science 76, 459–67.
Insam, H. (1990). Are the soil microbial biomass and basal
respiration governed by climatic regime? Soil Biology and
Biochemistry 22, 525–32.
Insam, H., and Haselwandter, K. (1989). Metabolic quotient of
soil microflora in relation to plant succession. Oecologia
79, 174–8.
Insam, H., Mitchell, C. C., and Dormaar, J. F. (1991).
Relationship of soil microbial biomass and activity with
fertilization and crop yield of three ultisols. Soil Biology
and Biochemistry 23, 259–64.
Insam, H., Parkinson, D., and Domsch, K. H. (1989). Influence
of macroclimate on soil microbial biomass. Soil Biology
and Biochemistry 21, 211–21.
Jenkinson, D. S. (1988). Determination of microbial biomass
carbon and nitrogen in soil. In ‘Advances in Nitrogen
Cycling in Agricultural Ecosystems’. (Ed. J. R. Wilson.)
pp. 368–86. (CAB International: Wallingford, UK.)
Jenkinson, D. S., Davidson, S. A., and Powlson, D. S. (1979).
Adenosine triphosphate and microbial biomass in soil. Soil
Biology and Biochemistry 11, 521–7.
Jenkinson, D. S., and Powlson, D. S. (1976). The effects of
biocidal treatments on metabolism in soil. V. A method for
measuring soil biomass. Soil Biology and Biochemistry 8,
209–13.
Joergensen, R. G. (1996). The fumigation–extraction method to
estimate soil microbial biomass: calibration of the k EC
factor. Soil Biology and Biochemistry 28, 25–31.
Karlen, D. L., Mausbach, M. J., Doran, J. W., Cline, R. G.,
Harris, R. F., and Schuman, G. E. (1997). Soil quality:
a concept, definition and framework for evaluation. Soil
Science Society of America Journal 61, 4–10.
Keeney, D. R. (1982). Nitrogen availability indices.
In ‘Methods of Soil Analysis’. Part 2. (Eds A. L. Page et al.)
pp. 711–33. (American Society of Agronomy: Madison,
Wisconsin, USA.)
Kuperman, R. G., and Carreiro, M. M. (1997). Soil heavy metal
concentrations, microbial biomass and enzyme activities in
a contaminated grassland ecosystem. Soil Biology and
Biochemistry 29, 179–90.
664
Ladd, J. N., Amato, M., Li-Kai, Z., and Schultz, J. E. (1994).
Differential effects of rotation, plant residue and nitrogen
fertiliser on microbial biomass and organic matter in an
Australian Alfisol. Soil Biology and Biochemistry 26,
821–31.
Lynch, J. M., and Panting, L. M. (1982). Effects of seasons,
cultivation and nitrogen fertiliser on the size of the soil
microbial biomass. Journal of the Science of Food and
Agriculture 33, 249–52.
Martens, R. (1995). Current methods for measuring microbial
biomass C in soil: potentials and limitations. Biology and
Fertility of Soils 19, 87–99.
McGill, W. B., Cannon, K. R., Robertson, J. A., and Cook, F. D.
(1986). Dynamics of soil microbial biomass and water
soluble carbon in Breton L after 50 years of cropping to two
rotations. Canadian Journal of Soil Science 66, 1–19.
McLaughlin, M. J., Alston, A. M., and Martin, J. K. (1988a).
Phosphorus cycling in wheat–pasture rotations. 2. The role
of the microbial biomass in phosphorus cycling.
Australian Journal of Soil Research 26, 333–42.
McLaughlin, M. J., Alston, A. M., and Martin, J. K. (1988b).
Phosphorus cycling in wheat–pasture rotations. 3. Organic
phosphorus turnover and phosphorus cycling. Australian
Journal of Soil Research 26, 343–53.
Myrold, D. D. (1987). Relationship between microbial biomass
nitrogen and a nitrogen availability index. Soil Science
Society of America Journal 51, 1047–9.
Oberson, A., Friesen, D. K., Morel, C., and Tiessen, H.
(1997). Determination of phosphorus released by
chloroform fumigation from microbial biomass in high P
sorbing tropical soils. Soil Biology and Biochemistry 29,
1579–83.
Pankhurst, C. E. (1994). Biological indicators of soil health and
sustainable productivity. In ‘Soil Resilience and Sustainable
Land Use’. (Eds D. J. Greenland and I. Szabolcs.)
pp. 331–51. (CAB International: Wallingford, UK.)
Parton, W. J., Schimel, D. S., Cole, C. V., and Ojima, D. S.
(1987). Analysis of factors controlling soil organic matter
levels in Great Plains grasslands. Soil Science Society of
America Journal 51, 1173–9.
Paul, E. A. (1984). Dynamics of organic matter in soils. Plant
and Soil 76, 275–85.
Perucci, P., and Scarponi, L. (1994). Effects of the herbicide
imazethapyr on soil microbial biomass and various soil
enzymes. Biology and Fertility of Soils 17, 237–40.
Powlson, D. S., Brookes, P. C., and Christensen, B. T. (1987).
Measurement of soil microbial biomass provides an early
indication of changes in total soil organic matter due to straw
incorporation. Soil Biology and Biochemistry 19, 159–64.
Powlson, D. S., and Jenkinson, D. S. (1981). A comparison of
the organic matter, biomass, adenosine triphosphate and
mineralisable nitrogen contents of ploughed and direct-
drilled soils. Journal of Agricultural Science 97, 713–21.
Rice, C. W., Moorman, T. B., and Beare, M. (1996). Role of
microbial biomass carbon and nitrogen in soil quality.
In ‘Methods for Assessing Soil Quality’. Soil
Science Society of America Special Publication 49.
(Eds J. W. Doran and A. J. Jones.) pp. 203–15. (Soil
Science Society of America: Madison, Wisconsin, USA.)
Robertson, F. A., Myers, R. J. K., and Saffigna, P. G. (1997).
Nitrogen cycling in brigalow clay soils under pasture and
cropping. Australian Journal of Soil Research 35,
1323–39.
R. C. Dalal
Saggar, S., Bettany, J. R., and Stewart, J. W. B. (1981).
Measurement of microbial sulphur in soils. Soil Biology
and Biochemistry 13, 493–8.
Sarathchandra, S. U., Perrott, K. W., and Littler, R. A. (1989).
Soil microbial biomass: influence of simulated temperature
changes on size, activity and nutrient content. Soil Biology
and Biochemistry 21, 987–93.
Schnurer, J. M., Clarholm, M., and Roswall, T. (1985).
Microbial biomass and activity in an agricultural soil with
different organic matter contents. Soil Biology and
Biochemistry 17, 611–18.
Seitz, L. M., Sauer, D. B., Burroughs, R., Mohr, H. E., and
Hubbard, J. D. (1979). Ergosterol as a measure of fungal
growth. Phytopathology 69, 1202–3.
Shen, S. M., Pruden, G., and Jenkinson, D. S. (1984).
Mineralisation and immobilisation of nitrogen in fumigated
soil and the measurement of microbial biomass nitrogen.
Soil Biology and Biochemistry 5, 437–44.
Singh, H., and Singh, K. P. (1993). Effect of residue placement
and chemical fertilizer on soil microbial biomass under
tropical dry cultivation. Biology and Fertility of Soils 16,
275–81.
Smith, J. L., and Paul, E. A. (1990). The significance of soil
biomass estimations. In ‘Soil Biochemistry’. Vol. 6.
(Eds J. M. Bollog and G. Stotzky.) pp. 357–96. (Marcel
Dekker: New York.)
Sorensen, L. H. (1983). The influence of stress treatments on
the microbial biomass and the rate of decomposition of
humified matter in soils containing different amounts of
clay. Plant and Soil 75, 107–19.
Spain, J. C., and van Veld, P. A. (1983). Adaptation of natural
microbial communities to degradation of xenobiotic
compounds: effects of concentration, exposure time,
inoculum, and chemical structure. Applied Environmental
Microbiology 45, 428–535.
Sparling, G. P. (1992). Ratio of microbial biomass carbon to
soil organic carbon as a sensitive indicator of changes in
soil organic matter. Australian Journal of Soil Research 30,
195–207.
Sparling, G. P. (1997). Soil microbial biomass, activity and
nutrient cycling as indicators of soil health. In ‘Biological
Indicators of Soil Health’. (Eds C. E. Pankhurst,
B. M. Doube and V. V. S. R. Gupta.) pp. 97–119. (CAB
International: Wallingford, UK.)
Sparling, G. P., and West, A. W. (1989). Importance of soil
water content when estimating soil microbial C, N and P by
the fumigation–extraction methods. Soil Biology and
Biochemistry 21, 245–53.
Sparling, G. P., Whale, K. N., and Ramsey, A. J. (1985).
Quantifying the contribution from the soil microbial
biomass to the extractable P levels of fresh and air-dried
soils. Australian Journal of Soil Research 23, 613–21.
Srivastava, S. C., and Singh, J. S. (1991). Microbial C, N
and P in dry tropical forest soils: effects of alternate land-
uses and nutrient flux. Soil Biology and Biochemistry 23,
117–24.
Staben, M. L., Bezdicek, D. F., Smith, J. L., and Fauci, M. F.
(1997). Assessment of soil quality in conservation reserve
program and wheat–fallow soils. Soil Science Society of
America Journal 61, 124–30.
 Microbial biomass: a review
Stockdale, E. A., and Rees, R. M. (1994). Relationship
between biomass nitrogen and nitrogen extracted by other
nitrogen availability methods. Soil Biology and
Biochemistry 26, 1213–20.
Tsai, C. S., Killham, K., and Cresser, M. S. (1997). Dynamic
response of microbial biomass, respiration rate and ATP to
glucose additions. Soil Biology and Biochemistry 29,
1249–56.
Valsecchi, G., Gigliotti, C., and Farini, A. (1995). Microbial
biomass, activity, and organic matter accumulation in soils
contaminated with heavy metals. Biology and Fertility of
Soils 20, 253–9.
Van Gestel, M., Merckx, R., and Vlassak, K. (1996).
Distribution of C-labelled biomass and C-labelled biomass
and microbial products in microaggregates of a silty-loam
soil. Soil Biology and Biochemistry 28, 1113–15.
Van Veen, J. A., Ladd, J. N., and Amato, M. (1985). Turnover
of carbon and nitrogen through the microbial biomass in a
sandy loam and a clay incubated with [14C] glucose and
[15N] (NH4)2SO4 under different moisture regimes. Soil
Biology and Biochemistry 17, 747–56.
Van Veen, J. A., Ladd, J. N., and Frissel, M. J. (1984).
Modelling C and N turnover through the microbial biomass
in a soil. Plant and Soil 76, 257–74.
Vance, E. D., Brookes, P. C., and Jenkinson, D. S. (1987). An
extraction method for measuring soil microbial biomass C.
Soil Biology and Biochemistry 19, 703–7.
Voos, G., and Groffman, P. M. (1997). Relationships between
microbial biomass and dissipation of 2, 4-D and dicamba in
soil. Biology and Fertility of Soils 24, 106–10.
View publication stats
665
Wardle, D. A., and Parkinson, D. (1990). Comparison of
physiological techniques for estimating the response of the
soil microbial biomass to soil moisture. Soil Biology and
Biochemistry 22, 817–23.
Webster, J. J., Hampton, G. J., and Leach, P. R. (1984). ATP in
soil: a new extractant and extraction procedure. Soil
Biology and Biochemistry 16, 335–42.
Wu, J., O’Donnell, A. G., He, Z. L., and Syers, J. K. (1994).
Fumigation–extraction method for the measurement of soil
microbial biomass-S. Soil Biology and Biochemistry 26,
117–25.
Zelles, L., Bai, Q. Y., Beck, T., and Beese, F. (1992). Signature
fatty acids in phospholipid and lipopolysaccharides as
indicators of microbial biomass and community structure in
agricultural soils. Soil Biology and Biochemistry 24,
317–23.
Zogg, G. P., Zak, D. R., Ringelberg, D. B., MacDonald, N. W.,
Pregitzer, K. S., and White, D. C. (1997). Compositional
and functional shifts in microbial communities due to soil
warming. Soil Science Society of America Journal 61,
475–81.
Received 27 November 1997, accepted 8 May 1998