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Soil microbial biomass—what do the numbers really mean?

Article  in  Australian Journal of Experimental Agriculture · January 1998


DOI: 10.1071/EA97142

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Volume 38, 1998
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Australian Journal of Experimental Agriculture, 1998, 38, 649–65 649

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 early changes induced by land use practices, zero
5% of organic matter in soil. However, it performs at tillage, crop rotations and other cultural practices,
least 3 critical functions in soil and the environment. It nutrient cycling, land disposal of sewage sludge, and
is a labile source of carbon, nitrogen, phosphorus, and applications of herbicides and insecticides. However, as
sulfur; it is an immediate sink of carbon, nitrogen, a routine analytical tool, it is limited by the
phosphorus and sulfur; and it is an agent of nutrient cumbersome and time consuming measurements, lack
transformation and pesticide degradation. In addition, of benchmarking values and interpretation, ambiguous
microorganisms form symbiotic associations with roots, relationship with productivity, and cost-effectiveness.
act as biological agents against plant pathogens, With increasing demand to monitor soil quality and
contribute towards soil aggregation, and participate in protection of the environment, improved and rapid
soil formation. techniques, including molecular biotechniques, will be
Critical evaluation of the significance of soil required to measure soil microbial biomass for its size
microbial biomass is hampered by the reliable of sink, source and rates of turnover. Eventually,
measurement of microbial biomass, and simultaneous agricultural science will benefit and utilise soil
partitioning of its 3 major functions in soil. For microbial biomass as an analytical tool to produce
comparative purposes, soil microbial biomass and its abundant, economical, and clean food and fibre.
derived indices have been successfully used to measure

Introduction nematodes) has been difficult because of incomplete


Soil microbial biomass is the living component of extraction, inappropriate growth media, extremely
soil organic matter. It excludes soil animals and plant variable growth habits of microorganisms in soil, and
roots. Although it comprises less than 5% of organic scant information on their biovolumes (Jenkinson 1988).
matter in soil, it performs at least 3 critical functions In the last 20 years, relatively rapid assessment of soil
for plant production in the ecosystem. It is a labile microbial biomass has been possible based on
source of carbon (C), nitrogen (N), phosphorus (P), and physiological, biochemical and chemical techniques
sulfur (S); it is an immediate sink of C, N, P and S; (Horwath and Paul 1994). These include chloroform
and it is an agent of nutrient transformation and fumigation incubation (CFI) (Jenkinson and Powlson
pesticide degradation. In addition, microorganisms form 1976), chloroform fumigation extraction (CFE) (Brookes
symbiotic associations with roots, act as biological et al. 1985; Vance et al. 1987), substrate-induced
agents against plant pathogens, contribute towards soil respiration (SIR) (Anderson and Domsch 1978), and
aggregation (Angers et al. 1992), and participate in soil adenosine triphosphate (ATP) analysis (Jenkinson et al.
formation. 1979; Eiland 1983; Webster et al. 1984). Of these, the
In the past, interest in microbial biomass assessment first 2 methods have been widely used to estimate
in soil has been limited by tedious, time consuming and microbial biomass in agricultural, pastoral and forestry
unreliable techniques (direct microscopy, culture media) systems, rehabilitation of disturbed lands, and pesticide
for enumeration of individual microbial communities. and heavy metals polluted lands and materials. Recently,
Estimation of microbial biomass from actual microbial biomass has even been proposed as a sensitive
enumeration of the whole suite of microorganisms indicator of soil quality (Karlen et al. 1997) and soil
(e.g. bacteria, fungi, actinomycetes, protozoa, health (Sparling 1997).

© CSIRO 1998 10.1071/EA97142


650 R. C. Dalal

But what do the soil microbial biomass numbers unfumigated (control) samples are extracted with 2 mol
really mean? I will examine this question and seek KCl/L after incubation. The mineral N in the extracts is
answers in the following sequence: (i) methods and their then determined colorimetrically or by steam distillation.
limitations to estimate microbial biomass; (ii) effects of As for microbial biomass C, a kN factor is used to correct
environment, soil, cultural practices and additives on the for incomplete mineralisation of N from killed
size of soil microbial biomass; (iii) source of nutrients microorganisms for calculating total biomass N.
and rate of mineralisation and its relationship to crop Soil microbial biomass C and N are calculated from
requirements; (iv) nutrient sink; (v) pesticide equations (1) and (2):
degradation; and (vi) as an indicator of soil quality.
Biomass C = (CO2-C fumigated – CO2-C control)/kC (1)
Methods to estimate soil microbial biomass Biomass N = (mineral N fumigated – mineral N control)/kN (2)
Detailed descriptions of methods and their limitations
to estimate soil microbial biomass by CFI, CFE, SIR and The widely accepted k C value is 0.41 at 22 o C
ATP analysis are given by Jenkinson (1988), Horwath (Anderson and Domsch 1978) or 0.45 at 25 o C
and Paul (1994) and Martens (1995). Both CFI and CFE (Jenkinson and Powlson 1976). However, kN varies from
methods measure a standing crop of soil microbial 0.30 to 0.68 (Smith and Paul 1990). Jenkinson (1988)
biomass, while SIR essentially measures microbial suggested a kN value of 0.57 at 25oC, which is about
activity and ATP is an indicator of microbial dynamics in 0.50 at 22oC.
soil. Chloroform fumigation incubation and CFE can Two basic assumptions of the CFI method are: (i) that
also be used to estimate N, P and S in soil microbial CO2-C evolved or mineral N produced during incubation
biomass and these can usually be used to recover added in fumigated soil must exceed that from the
radiotracers (e.g. 14C, 15N, 32P and 35S) from microbial corresponding unfumigated soil; and (ii) that CO 2-C
biomass. evolved or mineral N produced during incubation from
The microbial biomass of individual groups such as the non-microbial source must be equal in both
bacteria and fungi is estimated by microscopic methods fumigated and unfumigated soil samples (Jenkinson
and by measuring specific substances. The microbial 1988).
cell-wall components, muramic acid and glucosamine In soils with relatively low microbial biomass but
have been used as indicators of bacterial and fungal high respiration activity, subtraction of the CO2 evolved
biomass respectively (Atlas and Bartha 1981). However, from an unfumigated sample (control) often leads to low
microscopic techniques, and muramic acid and or even negative biomass estimates because unequal
glucosamine methods have difficulties in distinguishing amounts of non-microbial biomass C is mineralised
between living and non-living microbial biomass. The (Horwath et al. 1996). To overcome this problem,
cell-membrane-bound components such as phospholipid Jenkinson and Powlson (1976) suggested that CO2-C
fatty acids, have been used to estimate fungal and released during the 10–20 day incubation rather than that
bacterial biomass in soil (Zelles et al. 1992; Frostegard from the initial 0–10 day incubation of unfumigated soil
and Baath 1996). should be subtracted from the CO2-C released from the
Only a brief description and limitations of these fumigated soil. Horwath et al. (1996) suggested that the
methods are given below. proportion of CO2-C subtracted from the unfumigated
Chloroform fumigation incubation (0–10 day incubation) soil should vary as a function of
In this method, a moist soil is fumigated with ethanol- the ratio of CO2-C fumigated/CO2 control. When the
free chloroform for 24 h; chloroform is then removed by ratio is large the proportion of CO2-C subtracted from
repeated evacuation; the soil is reinoculated with a small the unfumigated soil should be large and vice versa.
amount of unfumigated soil and then incubated at a They also suggested that equation (1) can be modified to:
constant temperature (usually 22 or 25oC) for 10 days at Biomass C = (0.71 x CO2-C fumigated
field capacity or 50% of its water holding capacity – 0.23 x CO2-C control)/kC.
(about –0.01 MPa). An additional soil sample is retained
unfumigated and used as a control. The CO2 evolved However, the modified equation needs to be validated
during incubation can be measured by gas for soils under different land use and management and in
chromatography, as a continuous flow or by sorption in different climates.
alkali followed by titrimetric, conductometric or The 2 basic assumptions mentioned above do not hold
colorimetric determination. As the net C mineralised as for soils with pH <5, air-dried soils, waterlogged soils,
CO2 is only a proportion of the total microbial biomass and soils that contain recently added organic materials or
C, a kC factor is used to calculate total soil biomass C. plant residues. In acidic soils, the re-establishment of a C
For soil microbial biomass N determination, mineral and N mineralising microbial population after
N (NH 4 -N and NO 3 -N) from both fumigated and fumigation and reinoculation is very slow. This causes a
Microbial biomass: a review 651

reduced mineralisation of the killed microorganisms Limitations of this method are: (i) that the pattern of
which makes the usual k C and k N factors invalid soil microbial response to glucose differs between soils;
(Jenkinson 1988; Martens 1995). In air-dried soils, the (ii) that only glucose responsive soil microbial biomass
amount of already dead microorganisms may constitute is measured; (iii) that soils recently amended with
most of the microbial biomass in both fumigated and organic materials or plant residues contain a large
unfumigated soil samples, in addition to the less proportion of young cells, and, therefore, the conversion
effective lysing of microbial cells by chloroform factor used, from mL CO2/h to microbial biomass C of
(Sparling and West 1989). In waterlogged soils, CO2 and 40 (30 at 22 o C, Beck et al. 1997) for an average
CH4 are produced under conditions that restrict diffusion population in soil, is not valid (Martens 1995); (iv) it
of gases (Jenkinson 1988). In soils with recently added measures only microbial activity which does not
organic materials or plant residues, the second necessarily equate with microbial biomass; and (v) that
assumption is not met since the mass of the microbial biomass N, P and S cannot be measured
re-established microbial population in the fumigated and (Smith and Paul 1990).
reinoculated soil sample corresponds to only 10–20% of
the original microbial biomass and consists mainly of Adenosine triphosphate analysis
bacteria. This can be avoided by either careful removal Adenosine triphosphate is a universal constituent of
of the amendments such as roots, or a sufficient living microbial cells. Although ATP can occur in dead
preincubation of at least 3 weeks (Martens 1995). microbial cells and extracellularly in soil, it is rapidly
degraded by microorganisms. Therefore, ATP
Chloroform fumigation extraction
concentration in soil can be used to estimate the amount
The above-mentioned limitations of the CFI method
of living microbial biomass. It is usually extracted with
are mainly overcome by extraction of C and N with
acid reagents from moist, preincubated soil and
0.5 mol K2SO4/L from the chloroform fumigated and the
estimated by the luciferin–luciferase system. The C : ATP
unfumigated soil samples. The proportions of C (kEC)
ratio is about 200 although it varies from 120 to 240
and N (k EN ) extracted from the fumigated (killed
(Jenkinson et al. 1979; Eiland 1983; Martens 1995).
microbial biomass) soil vary from 0.2 to 0.68 (Jenkinson
The limitations of the ATP method are: (i) that ATP is
1988; Martens 1995). However, most frequently used
decomposed by enzymatic and chemical hydrolysis
kEC values are in the range 0.36–0.45, while the kEN
during the extraction process; (ii) after its release from
values are in the range 0.49–0.62.
Likely limitations of the CFE method are differential microbial cells, ATP is strongly sorbed by soil
extraction of released C from soils that differ in clay constituents (Martens 1995); (iii) biomass C : ATP ratio
content and clay mineralogy, and variable k values changes substantially over time in response to soil
(Martens 1995). amendments such as organic materials and plant residues
The CFE method has been successfully used to (Tsai et al. 1997); and (iv) it cannot measure microbial
estimate soil microbial biomass P (Hedley and Stewart biomass N, P and S in soil (Smith and Paul 1990).
1982) and S (Saggar et al. 1981). Inorganic P is
Phospholipid fatty acids
extracted with 0.5 mol Na2HCO3/L (pH 8.5) from both a Phospholipid fatty acids, with a chain length of <20 C
fumigated and an unfumigated soil; the proportion of P
atoms are considered to be of mainly bacterial origin
is extracted from the killed microbial biomass, and the kP
(Harwood and Russel 1984). However, 18-C chain
value is taken as 0.4. The allowance is also made for P
phospholipid fatty acid, 18 : 2ϖ6 fatty acid, constitute on
sorption during fumigation and extraction by including
average, 43% of the total phospholipid fatty acid in soil
an internal P standard. For strong P retention soils such
fungi (Federle et al. 1986). Since ergosterol is specific to
as Ferrosols, Bray extractant (30 mmol NH 4 F/L +
the fungal membrane (Seitz et al. 1979), the fungal
25 mmol HCl/L) appears to be more appropriate than
biomass can be estimated from the correlation between
0.5 mol Na2HCO3/L extractant (Oberson et al. 1997).
the amount of 18 : 2ϖ6 fatty acid and the ergosterol
The procedure for microbial biomass S determination
content. Frostegard and Baath (1996) observed a close
is similar to that for microbial biomass P. The most
correlation between the amounts of 18 : 2ϖ6 fatty acid
commonly used kS value is 0.41 (Smith and Paul 1990).
and the ergosterol in soil (r = 0.92), thus, indicating that
Substrate-induced respiration this phospholipid fatty acid can be used to estimate
An excess of substrate, usually glucose, is added to a fungal biomass. The ratio of 18 : 2ϖ6 fatty acid : bacterial
soil, which is then incubated at constant temperature and phospholipid fatty acid are then used as a
moisture, and the respiration rate, CO2 evolved per hour, fungal : bacterial biomass ratio (Frostegard and Baath
is measured during a 0.5–2.5 h period, before the 1996). Phospholipid fatty acids can be extracted from
microorganisms start proliferating and actually increase soil with a one-phase mixture of chloroform, methanol
microbial biomass. 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 R. C. Dalal

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

CO2

Residues Microbial
NOM
biomass

Ka Kd

POM
X

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).
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 657

incubation varies from 1 to 40 weeks (using either a transform non-biomass labile N into a mineral form for
static or a leaching procedure), and that of anaerobic the plant needs.
incubation from 1 to 4 weeks, usually at 30 or 40oC. However, it is uncertain whether the microbial
Because of this variability in the procedures used, it is biomass provides potentially mineralisable N through its
difficult to equate the reported values of potentially reduction in size or by mineralising labile organic N
mineralisable N to that of microbial biomass in soil. In fraction during a growing season. Turnover rates of
many instances only associated relationships can be microbial biomass range from 50 to 2000 days (Smith
established. and Paul 1990) while gross N transformation rates
Adams and Attiwill (1986) hypothesised that most of by and through microbial biomass are an order of
the NH 4 -N released during anaerobic incubation is magnitude faster (Shen et al. 1984).
derived from mineralisation of N from dead aerobic Measuring these pools as well as their rates of
microorganisms killed by anaerobic conditions. Myrold transformation is critical in accounting for the plant-
(1987) carried out anaerobic incubation (40oC, 7 days) in available N pool in soil. This is further complicated by
soils containing 15N-labelled microbial biomass. He the fact that during rapid plant growth, the size of the
found that the amount of NH4-N mineralised during microbial biomass may even increase or remain similar
anaerobic incubation was closely correlated with that due to C supply from root exudates (Dinwoodie and
mineralised in the CFI method. The equation describing Juma 1988), and thus it may compete with plants for N,
this was: albeit for a small amount (Robertson et al. 1997).
Fortunately, for most arable crops, the competitive
Anaerobic-N = 23 + 0.99 CFI-N (R2 = 0.81, P<0.01) (4)
demand for N by microbial biomass is small except
Dalal et al. (1991) also obtained similar results on a fine- when fresh plant residues have recently been added to
textured soil subjected to different crop residue soil.
management, fertiliser rates and tillage practices for
20 years. The equation was: Microbial biomass contribution to available phosphorus
Anaerobic-N = –8.2 + 0.97 CFI-N (R2 = 0.52, P<0.01) (5) and sulfur
The potential contribution of microbial biomass P to
However, it appears that not all N mineralised during plants may be assessed from the increase in available P
the anaerobic incubation is derived from microbial in soil when the soil is dried, presumably from killed
biomass N (equations 5 and 6). Bonde et al. (1988) microbial biomass. For example, Sparling et al. (1985)
found that only 55–89% of N released during anaerobic showed a good agreement in pasture soils between the
incubation was derived from microbial biomass. This increase in available P and the size of microbial biomass
was corroborated by Stockdale and Rees (1994). They P when the soils were air-dried (Table 8). The
also observed that microbial biomass N constitutes a contribution of microbial biomass P to available P varied
variable, although significant, part of the potentially from 4 to 76%. It was greater in soils containing greater
mineralisable N in soil. amounts of organic matter (>2% organic C). This
The amount of potentially mineralisable N measured phenomenon of varying amounts of microbial biomass P
over 40 weeks of incubation far exceeds the amount of mineralised during air-drying of soils has implications
microbial biomass N (6–10% v. 2–6% of total N) in soil for soil testing for available P in the laboratory. Since it
(Dalal and Mayer 1987; Bonde et al. 1988). Apparently, is not practical to use routinely moist soil samples for the
a substantial amount of non-biomass N but labile N is determination of available P, a standard procedure must
also mineralised during the long period of incubation. be used in air-drying soils for interpretations of available
Rice et al. (1996) concluded that the relationship P levels across regions and soils.
between microbial biomass and mineralisable N can be However, it is not certain whether microbial biomass
influenced by N fertilisation history and the contribution P released during drying of the soil may still be available
of non-living or dormant microbial biomass to N to plants when the soil is wetted. In fact, McLaughlin
mineralisation. Bonde et al. (1988) estimated that et al. (1988a) observed that the amounts of microbial
67–78% of total microbial biomass consist of a dormant biomass P in soil increased linearly with increasing soil
inactive fraction, which could be potentially mineralised water content in the field, with a large and rapid increase in
during the incubation period. Potentially mineralisable N the amount of P incorporated into the microbial biomass in
also increases with the addition of N through fertiliser, the first 7 days after wetting the initially dry soil.
plant material or farmyard manure, which enters the McLaughlin et al. (1988b) also found that the amount
labile fraction of soil organic N (Bonde et al. 1988). of microbial biomass P (27.9 kg P/ha) in a solonetz soil
The implication is that the role of microbial biomass was almost 4 times the amount that was taken up by the
N lies not only in supplying N from its biomass during wheat crop (7.5 kg P/ha) in 95 days. However, there is a
plant growth, especially after dry–wet cycles, but also to scarcity of information on microbial biomass P turnover
658 R. C. Dalal

Table 9. Microbial biomass phosphorus, microbial biomass sulfur, soil.day and 0.04 mg S/kg soil.day for arable soils while
available phosphorus and available sulfur in a clay loam pasture for grassland soils the values were 2-fold higher.
soil under different treatments (from He et al. 1997) However, these rates would include both the microbial
Treatments since 1897 include N applied at 35 kg N/ha.year biomass as well as non-microbial N, P and S sources in
(+ 19 kg S/ha.year), P at 26 kg P/ha.year (+ 2 kg S/ha.year), soil.
NPK (nitrogen, phosphorus, potassium) include N and P at the above
The average rate of microbial biomass C
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
mineralisation is usually taken as 2 x 10–3/day (Smith
NPK rates and Paul 1990); a value similar to the maintenance
Biomass P and available P were extracted by 0.03 mol NH4F/L + energy requirement for dormant organisms
0–025 mol HC/L, and kP value (proportion of P extracted from killed (2–4 x 10–3/day, Anderson and Domsch 1985). However,
microbial biomass) of 0.4 the individual N, P and S mineralisation rates of
Biomass S and available S were extracted by 0.01 mol CaCl2/L, and kS microbial biomass vary with the C : N, C : P, C : S ratios,
value (proportion of S extracted from killed microbial biomass) of 0.31 moisture, temperature, pH and soil matrix (Smith and
(Wu et al. 1994) Paul 1990).
Treatment Biomass P Available P Biomass S Available S
(mg/kg soil) (mg/kg soil) (mg/kg soil) (mg/kg soil) Limitations of microbial biomass values as the
indicators of nutrient source
Control 37 0.4 19 2.8 Although the amount of microbial biomass in a soil is
N 11 0.7 17 15.5 a potential source of nutrients to a crop, the
P 154 10.6 24 5.1 interpretation of its actual contribution to the crop needs
NPK 88 15.5 25 6.9 is complicated by the different microbial biomass
FYM 193 129 27 5.7 measurement techniques used, and the variables such as
FYM + NPK 178 172 18 6.3
moisture, temperature, pH, soil matrix, and
carbon : nutrient ratios that simultaneously affect
and management practices that can influence both the mineralisation–immobilisation processes, and
size and rate of turnover of microbial biomass P in soil competitive nutrient demands. Moreover, P and S
(Smith and Paul 1990; Horwath and Paul 1994). sources and availability are also controlled by abiotic
In pasture soils, biomass P and biomass S constitute a factors (Smith and Paul 1990).
substantial source of available P and S, especially where
soil fertility has been enhanced by regular applications of Microbial biomass as a sink of nutrients
fertilisers and organic manures (Table 9). He et al. Grasslands, woodlands and forests contain substantial
(1997) found that the amount of P in the pasture herbage amounts of microbial biomass. For example, the average
was more closely related to biomass P (R2 = 0.83) than microbial biomass pool size is 20–50% higher in
to the available P (R2 = 0.49) present in the soil, although grasslands and forest soils than in arable soils (Table 1),
the most significant correlation was found between the mainly because of the larger organic C inputs in the
amount of P in the herbage and the sum of both biomass former. Addition of easily metabolisable C to soil rapidly
P and available P in the soil (R2 = 0.98). increases microbial biomass, often by several fold,
Microbial biomass S as well as available S, however, and also incorporates nutrients into microbial
were poorly correlated with the herbage S yield, biomass rapidly, often within a week of addition
presumably because 0.01 mol CaCl 2 /L was a poor (Robertson et al. 1997; McLaughlin et al. 1988a).
extractant of S available over the whole season of However, such a microbial pool size is transient, and
herbage growth (He et al. 1997). the level of microbial biomass a soil can sustain under a
It should be emphasised that studies on the relative steady-state condition depends on the protective capacity
contribution of P and S from microbial biomass to the of the soil, which is governed by a dynamic interaction
available pool and then to the plant are complicated by between clay (Sorensen 1983; Van Veen et al. 1985), and
simultaneous chemical and physical processes such as clay aggregates (Van Gestel et al. 1996), soil microbial
sorption, diffusion into aggregates, occlusion and biomass and soil organic matter (Hassink and Whitmore
precipitation operating on P and S mineralised from the 1997), and possibly climate (Insam and Haselwandter
microbial biomass, and P uptake by the plant. 1989; Insam 1990).
Limited information is available on rates of Different management practices also affect the levels
transformation of microbial biomass of P and S. Smith of microbial biomass in a soil (Table 6). For example,
and Paul (1990) summarised the rates of N, P and S Schnurer et al. (1985) reported more than a 100%
mineralisation: the respective average rates of increase in microbial biomass C and N by the addition of
mineralisation were 0.5 mg N/kg soil.day, 0.02 mg P/kg 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 manure (205 mg C/kg soil) over the same period. As the
biomass P values increased by 2–5-fold with the EDTA-extractable nickel and copper (Cu) increased,
application of P fertilisers and manures for over 90 years microbial biomass decreased, and these effects were
(Table 9); however, these observations need to be detectable even after 20 years of application.
confirmed in different environments. Furthermore, Chander and Brookes (1993) found that a
Limitations of microbial biomass values indicating the combination of zinc (Zn) and Cu at high concentrations
nutrient sink source had an additive adverse effect on the amount of
Carbon flux through microbial biomass is the driving microbial biomass present although, at similar
force for the decomposition of plant residues and other concentrations (CaCl2-extractable Cu and Zn, 415 and
457 mg/kg respectively), microbial biomass decreased
organic materials. While microbial biomass is building
by 50% more with Cu than Zn (Table 10).
up it can immobilise substantial amounts of N, P and S.
However, eventually C becomes limiting, microbial Also, the microbial quotient decreases much faster
biomass dies, and a large proportion of N, P and S is than the total microbial biomass as the heavy metal
transferred to the non-living microbial biomass. Thus, concentration increases. For example, the microbial
microbial biomass values, at the most, indicate only the quotient decreased from 1.5% in soil that received
partial source of nutrients for a crop. Moreover, uncontaminated sewage sludge to 1% with Zn
limitations concerning the variability in procedures of (457 mg/kg soil) and 0.5% with Cu (415 mg/kg soil)
measuring microbial biomass, and the variables affecting contained in the sludge applied (Table 10). Valsecchi
it still apply. et al. (1995) found that large additions of heavy metals
through long-term effluent decreased the microbial
Effects of heavy metals and pesticides quotient more than an order of magnitude, from 4% in
Increasing disposal of sewage sludge on land has the the slightly contaminated soil to 0.2% in heavily
potential to increase heavy metals in soil. There is thus contaminated soil; the ratio of specific respiratory rate to
concern about the long-term effects of additions of heavy organic C decreased only by 50% (Table 11). On the
metals in sewage sludge to soil. It is believed that other hand, both the specific respiration rate and
microbial biomass, being the living component of soil, respiratory quotient were much higher in the latter.
can provide an early indication of the adverse effects Thus, the microbial quotient provides a sensitive
of increasing concentrations of heavy metals in indicator of the effects of heavy metals on soil microbial
soil (Brookes and McGrath 1984; Chander and biomass. This may be because either the soil microbial
Brookes 1993; Brookes 1995; Valsecchi et al. 1995; population becomes smaller (Brookes and McGrath
Dahlin et al. 1997). 1984) or soil microbial biomass becomes less effective
Brookes and McGrath (1984) found that the amounts in mineralising soil organic matter, that is, turnover rate
of microbial biomass in agricultural soils treated with of organic matter decreases (Valsecchi et al. 1995) as
either sewage sludge or sludge-containing composts organic matter accumulates in soil with the applications
(average microbial biomass, 93 mg C/kg soil) were of the heavy metals contaminated sewage sludge.
much smaller than in soils which received farmyard The higher respiratory quotient, combined with the
660 R. C. Dalal

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 MBC SRR MBC/organic C SRR/MBC SRR/organic C
Zn Cu Pb Ni Cr Cd (%) (mg C/kg) (mg CO2-C/kg.day) (%) (%/day) (%/day)

1 17.9 12.2 9.2 1.6 2.7 0.6 0.90 366 3.66 4.06 1.01 0.04
2 16.9 16.2 16.7 2.5 3.7 0.7 1.95 555 8.99 2.85 1.62 0.05
3 4884.0 228.0 771.0 24.0 30.0 41.0 5.60 125 11.48 0.22 9.18 0.02

lower ratio of specific respiratory rate to organic C, in benomyl, chlorfenvinphos, glyphosate, chlorotoluron or
the heavy metals contaminated soil indicates a state of triadimefon to soil, used for barley cropping, for up to
microbial stress due to heavy metals toxicity. These 20 years.
metals reduce the energy efficiency of the microbial The application of fungicides shifts the balance of
metabolic processes, which then require greater amounts microbial biomass towards bacteria-dominated microbial
of C for maintenance, thus reducing the quantity of C biomass (Spain and van Veld 1983). We need to know a
incorporated into the microbial biomass (Valsecchi et al. great deal about the effects of herbicides and pesticides,
1995). and other organic additives, especially under repeated
Since the toxic effects of heavy metals on microbial applications on the 3 functions of microbial biomass in
biomass are additive, it has been suggested that even soil: as a source and a sink of nutrients, and an agent of
smaller concentrations of a combination of heavy metals transformation of nutrients.
in sewage sludge may affect not only microbial biomass
and activity but also community structure as determined Limitations of microbial biomass values as indicators of
by phospholipid fatty acids analysis (Dahlin et al. 1997). heavy metal toxicity
This is expected since heavy metals such as Cu Microbial biomass values as such do not usually
adversely affect the fungal community. However, limited provide good indicators of heavy metal toxicity in soil.
information is available on the effects of heavy metals Some enzyme activities (Kuperman and Carreiro 1997)
on nutrient cycling by and through microbial biomass and susceptible plants are much more sensitive
although N and P hydrolysing enzymes have been found indicators. However, some microbial indices such as the
to be adversely affected (Kuperman and Carreiro 1997). microbial quotient or the ratio of specific respiratory
A substantial proportion of cropping land receives rate/organic C may prove useful.
regular applications of herbicides and insecticides. While
herbicides applied at the field rate are decomposed in Microbial biomass as an indicator of soil quality
soil with only a slight effect on the amount of microbial Soil quality is the capacity of a specific kind of soil to
biomass (Perucci and Scarponi 1994; Voos and function, within natural or managed ecosystem
Groffman 1997), some insecticides applied to soil not boundaries, to sustain plant and animal productivity,
maintain or enhance water and air quality, and support
only reduce the size of microbial biomass, but also alter
human health and habitation (Karlen et al. 1997). This
the composition of the microbial communities (Spain
definition should be extended to include ‘maintain or
and van Veld 1983; Bromilow et al. 1996). enhance soil biodiversity’. To evaluate soil quality, the
Perucci and Scarponi (1994) found that the herbicide, capacity of the soil to function needs to be measured
imazethapyr (amidazolinone derivative) used to control a using appropriate indicators. The most desirable
wide spectrum of broadleaf weeds and grasses, when attributes of an appropriate indicator include the
applied at recommended field rate (50 g a.i./ha) had no following: (i) it measures one or more soil functions;
effect on soil microbial biomass although at 100-fold the (ii) it is sensitive enough to measure changes due to
field rate it had an adverse effect on the amount of disturbance, restoration, or change in land use and
microbial biomass. Similar results were obtained by management; (iii) it provides benchmark, critical or
Voos and Groffman (1997) for the microbial breakdown threshold values; (iv) it can be readily interpreted; and
of 2,4-D and dicamba. They also observed that the rate (v) it is cost-effective.
of dissipation of the herbicides increased with the Microbial biomass performs 3 essential functions in
amount of microbial biomass in the soil. Bromilow et al. soil (see ‘Introduction’). It is also sensitive enough to
(1996) also measured no adverse effects on the amount measure the changes due to the different land use and
of microbial biomass of annual application of aldicarb, management such as virgin land brought under
Microbial biomass: a review 661

cultivation (Dalal and Mayer 1987), crop rotations common aspect of these 2 systems appears to be the low
(McGill et al. 1986), restoration (Staben et al. 1997) and incorporation of C into microbial biomass, and by
heavy metal contamination (Brookes and McGrath inference, probably leading to low soil microbial
1984). However, currently, microbial biomass does not diversity.
provide benchmark, critical or threshold values against The ratio of fungal biomass to bacterial biomass
which soil quality can be evaluated. appears to be sensitive to land use changes. For example,
The benchmark values will differ depending on soil this ratio was found to be higher in a restorative soil
type and land uses (Sparling 1997), climate (Insam et al. system than in the cultivated soil (Staben et al. 1997).
1989), and vegetation (Martens 1995). Such differences This was a reflection of the dominance of the bacteria-
can be reduced by comparing the derived microbial feeding nematodes in the former; which had a ratio of
indices such as the microbial quotient for soils of fungal-feeding nematodes to bacterial-feeding
different organic matter contents (Powlson and nematodes of 0.5 compared with 4.8 for the cultivated
Jenkinson 1981; Anderson and Domsch 1989; Sparling soil. However, the applicability of microbial diversity to
1992) and respiratory quotient for soils of different soil quality requires further study although its cost-
microbial biomass contents (Insam and Haselwandter effectiveness may limit its measurement as a routine
1989; Anderson and Domsch 1990). In addition, the measure of soil quality.
ratio of microbial-specific respiration rate/organic C has
Limitations
been proposed as a sensitive indicator for heavy metal
Besides the current limitations of unavailability of
contaminated soils (Valsecchi et al. 1995).
benchmark values and difficulty in interpretation, soil
The benchmark values of the microbial biomass and
microbial biomass measurements need to be cost-
even the derived microbial indices for soil quality are not
effective. Moreover, current methods of measuring
available (Sparling 1997) although the differences in
microbial biomass, its activity, and biodiversity in soil
both microbial biomass and microbial indices can be
are laborious, cumbersome and suffer from a lack of
demonstrated for different soil types and land uses and
close association with productivity. Although microbial
management (Tables 1, 2, 5–11). Do such values exist?
measurements are essential in evaluating soil quality,
Are these values available in the literature on soil
these limitations need to be overcome if soil microbial
microbial biomass? With our current understanding of
biomass is to be routinely measured and used as an
soil microbial biomass and soil quality, the answer is not
indicator of soil quality.
in the affirmative. It is tempting to suggest that the
(maximum) potential amount of microbial biomass in a Future perspectives
soil could be estimated from the protective capacity of a Smith and Paul (1990) asked the question, ‘What do
soil (Hassink and Whitmore 1997) for the specified our microbial biomass values represent and how can we
climate. By corollary with organic matter, the potential use them to study nutrient cycling?’
microbial quotient can then be benchmarked. Since 1990, microbial biomass measurements have
However, the problem of interpretation of the been extensively used to study nutrient cycling. We
benchmark values and deviation from these values in know that although microbial biomass is only a small
terms of soil quality is still unresolved. It is tempting to component of soil organic matter, usually <5%, nutrient
suggest that the extreme stress on the soil’s ecosystem fluxes through microbial biomass are at least an order of
can be identified from a combination of the derived magnitude faster than the remaining organic matter;
microbial indices (Table 11). From Tables 10 and 11, it is hence, its study is critical in understanding the nutrient
suggested that the microbial quotient value of <0.5 could cycling in soil.
be considered an indicator of the soil under stress (poor Due to the rapid turnover rate of microbial biomass, it
soil quality, that is, the impairment of the soil’s capacity responds to changes in soil and crop management
to function). Further work is required to validate this practices much faster and earlier than the bulk of soil
value for soil microbial biomass under stress due to organic matter. Therefore, microbial biomass
factors other than heavy metal contamination. measurements have been critical in monitoring early
On the other hand, the significance of specific changes in soil induced by zero tillage practice,
respiratory rate and respiratory quotient on soil quality is restorative practices, use of herbicides and insecticides,
less clear. For example, both highly productive soils as and application of sewage sludge to land.
well as contaminated soils exhibit high respiratory rates For a given soil and cultural practice, soil microbial
and high respiratory quotients (Brookes and McGrath biomass has been useful as an indicator of changes in
1984; Alvarez et al. 1995; Valsecchi et al. 1995; soil due to the above-mentioned practices. Therefore, it
Sparling 1997) although the ratio of specific respiration has been suggested as an important biological indicator
rate to organic C should be higher in the former. How do of soil health (Pankhurst 1994; Sparling 1997) and soil
these measurements relate to the soil quality? The quality (Karlen et al. 1997). However, as a management
662 R. C. Dalal

tool its applications have been limited because Anderson, T. H., and Domsch, K. H. (1989). Ratios of
analytically soil microbial biomass needs to have microbial biomass carbon to total carbon in arable soils.
measurements of all of its 3 functions simultaneously: Soil Biology and Biochemistry 21, 471–9.
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