International Journal of Agricultural

Science and Research (IJASR)

ISSN (P): 2250-0057; ISSN (E): 2321-0087
Vol. 4, Issue 6, Dec 2014, 1-10
TJPRC Pvt. Ltd.

EFFECT OF PRESCRIBED FIRE ON SOIL MICROBIAL BIOMASS IN A

MEDITERRANEAN FOREST (PINUS HALEPENSIS) ECOSYSTEM AROUND
MONTPELLIER CITY, SOUTH OF FRANCE
AHMED FAHD RACHID
Department of Soil Science and Water Resources, College of Agriculture, Diyala University, Diyala, Iraq

ABSTRACT
This work was conducted to study soil microbial biomass (MB) changes concerning Aleppo pine
(Pinus halepensis) forest ecosystem, located in the south of France bordering Montpellier city, subjected to a prescribed
fire on 30 March 2011. Soil MB and moisture content were measured immediately before and after burning; experiment
monitoring lasted five consecutive months. Within 24 hours, MB significantly increased by 26% in soil surface (0-2 cm).
This achieved level of MB pursued continuously and significantly until soil moisture has reached 51% after a month of
heavy rains which unexpectedly reduced MB size to 12% and 31% from its initial level before burning, in burned sites and
control, respectively. From mid-May till the end of July, MB once again gained its initial concentration and its levels were
higher in burned sites, but sometimes not significantly. At 2-10 cm soil layer, tendencies of MB changes are similar to
those of soil surface but increases often were not significant. However, two significant and highly significant increases
occurred when they are immediately preceded by periods of saturation state of the above soil surface where humidity
ranged between 47% and 54% from mid-April to mid-May. Our results indicate first that when pine forest ecosystem is
moderately moist, prescribed fire shows a low intensity and subsequent increases occur in soil MB. Although these
increases were not frequently significant, they were more or less lasting until the following season; and secondly that the
seasonal changes in water availability especially those leading to soil anaerobic conditions in Mediterranean region, is
considered an important determining factor for the MB.

KEYWORDS: Mediterranean Forest Ecosystem, Moisture Content, Prescribed Fire, Soil Microbial Biomass
INTRODUCTION
If wildfires are a destructive force and have adverse effects on ecosystems, prescribed burning can be a tool of
forest and agricultural management. Prescribed burning is known as the controlled use of fire to achieve specific forest
management objectives, e.g. reduction of fire hazard, control of competing vegetation, creation of seedbeds and planting
spots, and overall improvement of the efficiency of silvicultural operations by removing impediments to reforestation and
stand management. Two types of prescribed burning are used in forestry: under-burning, i.e. burning under mature forest
canopies, and slash burning, i.e. a method of disposing of logging residue (Walstad et al., 1990). In this study the term
prescribed burning refers to under-burning. Controlled fire effects on the soil environment require to be known because
they have direct implications on the soil surface layers, where are concentrated the organic matter, nutrients and biological
activity (growth of roots, microbial biomass, mineralization, etc.). These effects depend on fire severity which consists of
two components: intensity and duration (Hatten and Zabowski, 2009). Fire severity is controlled by several environmental
factors that affect the combustion process, such as amount, nature and moisture fuel, air temperature and humidity, wind

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Ahmed Fahd Rachid

speed, and soil environment (Liu et al., 2010). The passage of fire in the litter and the herbaceous layer or the lower woody
stratum affects both soil physical properties such as temperature, humidity, erosion, etc., and soil nutritional status such as
nutrient-rich ash provision (Zhao et al., 2010) or loss of essential nutrients such as nitrogen (Qian et al., 2009).
This combined action interferes itself on soil biological activity so that it frequently increases plant growth. In fact, by a
statistical analysis of data from 48 observations, Johnson and Curtis (2001) disentangled a positive long-term effect of fires
on the stock of soil organic carbon, which increased by 8% in average.
Nevertheless, assessment of soil organic carbon must be evaluated not only on a quantitative basis but also on a
qualitative basis such as soluble organic matter or microbial biomass (MB). Moreover, microbiologists have long believed
that the level of MB (Bacteria, Fungi, Virus, etc.) and its activity are major components of the concept of soil fertility;
more and more agronomic studies concerning the changes in soil organic matter stocks take explicitly into account the
microbiological aspects (Anderson, 2003; Akihiro Koyama et al., 2011). In addition, MB amount, which constitutes in
weight between 1 and 5% from total organic matter, represents a more sensitive indicator of soil process variations
although it has a faster turnover than total organic pool. However, many previous studies concerning prescribed burning
impact on soil MB showed conflicting results. From a meta-analysis of 42 published studies, Dooly and Treseder (2012)
reported an immediate reduction in soil MB as a whole, but they specified that MB response to fire differed significantly
among biomes and fire types. For example, MB declined following fires in boreal and temperate forests but not following
grasslands fires. In addition, wildfires lead to a greater reduction in MB than prescribed burns. Contradictorily, Ahlgren
(1974) explained that MB in Mediterranean ecosystem could be expected higher in the burned surfaces than in the
unburned. Moreover, other studies detect no change in MB after burning (Docherty et al., 2012).
This research aims to monitor the MB in the soil under a pine forest around Montpellier, France, in studying a
burned and a control sites immediately after a controlled fire. Then we will put in evidence the long-term impact of the five
months followed the burning. As well as we will explore the relationship between soil moisture and the resulting changes
in the MB size.

MATERIALS AND METHODS

Study Sites Description and Climate Conditions
A piece of forest land, at the experimental station of the Higher Agricultural Institute in the city of Montpellier,
southern France, has been divided in two plots (700 m2 each) identical in vegetation and soil characteristics. One plot was
treated by the fire and the other was left as control. Soil is a Mediterranean Fersiallitic on dolomitic limestone bedrock
from the Jurassic period (Duchaufour, 1965). The CaCO is in the form of pebbles and rocks in place. The texture is a
loam with 35.3% sand, 39.78% silt, and 24.92% clay. Soil pH values measured on soil water suspension (1: 2.5 ratio)
ranged from 7.8 to 8.3 and those measured from a suspension of KCl (1N) ranged from 7 to 7.3 in the control and the
burned areas, respectively. Soil pH practically does not vary during the experiment. The study sites are dominated by
Aleppo pine (Pinus halepensis) with a density of 3400 trees/ha. The other strata are distributed as follows: undergrowth is
dominated by Kermes oak (Quercus coccifera) and Rosemary (Rosmarinus officinalis) constituting 8.8 tone of dry
matter/ha; herbaceous cover is composed of prevailing weed, Bromine erected (Bromus erectus) and Brachypodium
branching (Brachipodium ramosum) representing 0.3 tone of dry matter/ha; fallen leaves and dead twigs (Litter layer)
covering 90-100% of the soil surface.

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effect of Prescribed Fire on Soil Microbial Biomass in a Mediterranean Fores

(Pinus halepensis) Ecosystem Around Montpellier City, South of France

The general climate of the region is sub-humid Mediterranean. The rain-heat ratio is 80; the average of the annual
temperature is 14.4 C. The average rainfall of the last 64 years was measured to 770 mm with a characteristic distribution
of the Mediterranean climate: rainfall is concentrated in the cold season of the year and vegetation is subject a period of
summer drought (Figure 1).

.
Figure 1: Rain-Thermal Readings of the Weather Station of C.N.R.S. Montpellier
Burning Experiment
Fire ignition was performed, on 30 March 2011, in the afternoon and lasted two hours, with drip torch by flaming
line front against wind direction. Punctual and/or stripping sources were used to increase fire intensity into a satisfactory
level, in case of any casual gap. The air temperature was 14.5 C and the relative humidity was 80% with 0.6 mm
precipitation on that day. The average wind speed was 6.5 m/sec and fire speed was from 20 to 25 m/hour. The average
height of the fire flames was 1.1 meters.The fire had slightly burned trunks and tops of smaller trees, and often consumed
litter and grass as well as the lower thin branches of trees.
Soil Sampling and Moisture Determination
Soil samples were collected at eleven appointments. The first two were immediately before and one day after
burning; the remaining sampling dates were at about 10 day intervals for the next three sampling,15 day intervals for the
following four sampling, and 30 day interval for the last sampling. Soil samples were taken at two depths 0-2 cm and
2-10 cm from both sites burned and control. Four specimens were objectively performed at each sampling. The soil was
screened through a 2 mm sieve to remove large roots, dead leaves, macro fauna, and stones. Then, it was homogenized and
left in two lots to evaluate the microbial biomass (MB) size in the soil: one used in fumigation and the second served as a
control non-fumigated, both followed by incubation. Samples were used in the fresh state after sieving to avoid impact of
desiccation on the microbial activity.
Soil moisture content was measured by gravimetric method. Fresh sample of soil was oven dried at 105 C until
reaching constant weight. Then, water content was calculated on the basis of fresh and dry weight difference and expressed
as percentage of dry weight. Soil Water holding capacity (WHC) was determined using the method described by
Duchaufour (1965).

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Ahmed Fahd Rachid

Microbial Biomass Assessment and Analysis

Microbial biomass carbon (MBC) was determined by the chloroform-fumigation incubation (CFI) method as
preconized by Jenkinson and Powlson (1976c) and modified by Chaussod and Nicolardot (1982). CFI technic measures the
total MB under optimum environmental conditions. The principle of this method is that the chloroform (CHCl3) kills
microbial cells and causes the lyses of its walls, providing easily decomposable carbon sources; after chloroform removing,
samples were reseeded with a small amount of fresh soil. The new introduced population and surviving microorganisms
after fumigation will mineralize microbial bodies with additional release of CO2 which measured after 10 days of
incubation in- vitro. This extra of CO2 "flush" from fumigated sample compared to a non-fumigated sample is proportional
to microbial biomass that would be found in the soil before fumigation. The CO2 released after 10 days of incubation from
the non-fumigated sample "flush less", represents the non-microbial origin CO2.
The difference in CO2 released from fumigated and non-fumigated soil samples between zero and ten days, allows
assessing the "flush" corresponding to the decomposed microbial bodies: flush = {fumigated (C-CO2
non-fumigated (C-CO2

0-10 d.inc)}.

0-10 d.inc)

The amount of MBC was then calculated using the following formula: MBC = flush /

KC. The proportionality factor (Kc), which is the coefficient between the total microbial carbon and microbial carbon
mineralization in the form of CO2 in the "flush", was commonly estimated to 0.41. Colorimetric determination was
performed to measure CO2 fixed by the soda according to the reaction: CO2+2NaOH
quantitative

colorimetric

chain

by

sulfuric

acid

H2SO4

(3.6N)

CO3Na2+H2O. It moved on the

according

to

the

reaction:

H2SO4+CO3Na2 CO2+Na2SO4+H2O. Then, CO2 released discolor solution phenolphthalein in buffer carbonate and
sodium bicarbonate (NaHCO3 and NaCO3). Discoloration of the solution of phenolphthalein, which is expressed as optical
density, is measured at 570 nm. Statistical analysis was applied on our experimental data, which is inversely proportional
to the optical density, using the program developed at CNRS Montpellier. The MBC of the two plots is determined from
eight trials. Average differences between the burned plot and the unburned are tested according to the Students't
distribution with a safety factor of 95% and 99%.

RESULTS AND DISCUSSIONS

The temperature in the litter under Kermes oak bushes, ranged from 174 to 260 C, but it did not exceed 71 C at
the upper thin layer of soil surface. It was less than 40 C on the soil surface below the litter in the herbaceous areas.
In general, the soil temperature at 0-2 cm depth varied between 40 and 60 C, but it did not increase at a depth of 2-10 cm.
Temperatures under Kermes oak layers related to fire residual time are shown in Figure 2.

Figure 2: Soil and Litter Layer Temperatures during Ecosystem Burning

Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effect of Prescribed Fire on Soil Microbial Biomass in a Mediterranean Fores

(Pinus halepensis) Ecosystem Around Montpellier City, South of France

One of the first observations is that the temperature gradient between the litter layer and soil layers is very steep.
This wide difference between temperatures is explained by the attenuating effect of 0.6 mm precipitation, the day of the
experiment, through two possible mechanisms; first, the organic matter conducts heat poorly and second, as long as its
moisture increases, its insulating capacity reinforces by latent heat effect (Zu et al., 2009). Although heat in moist soil is
transported faster and penetrates deeper, latent heat of vaporization prevents soil temperature from rising rapidly until
water completely vaporizes (Campbell et al., 1995). For this reason and despite precipitation, soil water content decreases
about 5% (from 15.18 to 10.63%) at 0-2 cm depth one day after burning. At the depth 2-10 cm, the temperature remains
lower than 40 C and soil water content decreases only by 1% (from 13.84 to 12.46%).
Second observation is that whatever depths and sampling periods, soil water content is higher in burned sites than
in control sites (Figure 3); over the ten sampling periods after burning, the mean increase was as much as 6%. As a
management practice, prescribed fire could prove soil water retention and subsequent resistance against drought of summer
months in Mediterranean basin region.

Figure 3: Burning Effect on Soil Moisture Content (%), At the 0-2 cm (Right) and 2-10 cm (Left) Layers, Over
Experiment Period
Soil temperature and humidity results combined with the measured values of weather data (Figure 1), registered
observations of burned strata, and fire behavior description in terms of flame length, spread rate, suggest, according to
Fernandes' and Loureiro's index (2010) concerning the optimal range of prescribed fire variables, that the type of our
prescribed fire was of low intensity. Consequently, soil sterilization and lethal effect on microorganisms, except for
Nitrobacter and some nitrifying as Nitrosomonas, are likely not reached. In fact, these species may be killed at 50 C and
75 C, respectively, in a moist soil (Dunn and Debano, 1977). In general, bacteria and actinomycetes are more resistant to
heat than fungi. The lethal temperature for bacteria is around 110 C in moist soil in Chaparral (Debano et al., 1979). Fungi
could persist up to 100 C temperature, point at which they are finally destroyed.
A day after burning, a significant increase (P = 0.05) in MB size occurred at the 0-2 cm of soil surface (Table 1)
and there was a slight non-significant increase at the depth of 2-10 cm (Table 2). Compared to the same sites just before
burning, these immediate increases were 26% and 16% respectively. Altogether, rewetting of the soil caused by rainfall on

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Ahmed Fahd Rachid

the day of the fire, combined with energetic (low heat) and trophic (nutrient-rich ash) flows could allow microorganisms to
grow and stimulate biodegradation of organic matter and available humic substances. This result is in accordance with the
data obtained by Banning and Murphy (2008) in eucalyptus forest soil exposed to a heat-induced disturbance at 60 C.
Also, Mataix-Solera et al. (2009) observed a short-term increase in microorganism populations due to the increase in
soluble carbon and nutrients after forest fire. Conversely, our data were inconsistent with those obtained after passage of
high intensity fire (Dunn et al., 1979; Dooly and Treseder, 2012; Brcenas-Moreno et al., 2011).
During the two weeks after the fire, the microbial biomass at the 0-2 cm level peaked in both plots burned and
control, but its size in burned plot was significantly higher (P = 0.01) than that of the control. The increase is directly
related to seasonal variations; microbial activity is very important in March and April.
Table 1: Changes in Soil Microbial Biomass Carbon (Mg / 100g. Dry Soil) and Water Content (%) at 0-2 cm Depth
MBC mg/100g.Dry Soil
Date
Control
30 / 3
Fire
date

222
(18)

Burned
234NS
(19)

Water Content (%)

Control
12.73

Burned
15.18

295*
ND
10.63
(12)
**
300
373
11 / 4
37.95
46.7
(22)
(12)
*
180
343
30.87
46
19 / 4
(68)
(31)
NS
68
29
02 / 5
42
50.7
(50)
(11)
NS
287
250
16 / 5
40.66
54.18
(94)
(73)
188
187NS
30 / 5
11.82
24
(19)
(12)
159
217*
13 / 6
6.28
13.18
(19)
(23)
196
193
27 / 6
5.98
8.44
(12)
(17)
*
197
233
3,35
4.54
25 / 7
(17)
(15)
5/8
6.82
8.40
ND
ND
NS
Each Value is the Average of Four Replicates. : Not-Significant Difference. *: Significant Difference At 95% of
Probability. **: Significant Difference At 99% of Probability. ( ): Standard Deviation. ND: No Determined
31 / 3

ND

One month after the fire, MB decreased sharply to a minimum (29mg/100g dry soil) in the burned plot where soil
moisture was 51%, and gradually (68mg/100g dry soil) in control where soil moisture was 42%; the MB is reduced to 12%
and 31% from the respective initial values in the burned area and the control (Table 1). This phenomenal reduction,
strongly linked to a phase of water saturation of the soil, is probably due to a hydrophobic character of soil surface. Indeed,
soil surface originally contains hydrophobic substances such as aliphatic hydrocarbons leached from litter layer
(Doerr et al., 2000). Furthermore, low to moderate fires impose, by partial pyrolysis, major chemical and structural
changes of all main organic matter components, forming more hydrophobic and insoluble compounds such as aromatic
substances (Gonzalez-Perez et al., 2004). These decomposed organic particles and the external hydrophobic surface of
aggregates would be coated with a discreet and continuous thin layer of "water repellent" that reduces soil permeability
(Letey, 2001; Mataix-Solera and Doerr, 2004). In a soil under pine forest, Robichaud (2000) measured 40% fire-induced
Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0

Effect of Prescribed Fire on Soil Microbial Biomass in a Mediterranean Fores

(Pinus halepensis) Ecosystem Around Montpellier City, South of France

reduction in hydraulic conductivity following slash burning.

Thus the new rainfall, occurred during mid-April to mid-May, did not penetrate easily deep into the soil.
This hyper humidity can either modify the gas exchange disadvantaging soil climatic conditions for the development of
aerobic microorganisms and simultaneously providing optimal biological activity of anaerobic conditions; anaerobic
environment may enhance the toxic action of water-soluble inhibitors, resulting from litter and microbial metabolism, of
microbial activity (Chen et al., 2008); or vertically and laterally transport various substances including energetic substrates
or/and elements of the micro flora such as vegetative cells or spores. Leaching of organic and mineral reserves of soil and
possible mechanisms of redox related to high humidity (Steven et al., 2013), are generally accompanied by a gradual
reduction of the MB even outside the dry season (Wood et al., 2013). Finally, changing in soil environment necessarily
results in soil microorganisms diversity changes. However, whatever fire severity and soil features, "water repellent" soil
layer rarely exceeds 3-6 cm in depth and shows weakening 3 months after fire (Huffman et al., 2001).
From mid-May till mid-June, the MB raised again to levels similar to baseline levels before burning. For this
period, the differences between the burned area and the control were non-significant, except for the sampling of mid-June.
Similar results were found for the period going until the end of July, which ascertain that contrary to the control, the
prescribed burning improves significantly soil resistance to summer drought.
At the 2-10 cm soil layer, levels of MB in burned sites often remain not significantly higher than those of the
control sites (Table 2). MB increased for the first two weeks after the fire (189 and 140mg/100g of dry soil in the burned
area and the control respectively). Then it decreased to a minimum at the beginning of May but later it increased to a
maximum of 216mg/100g dry soil for the burned plot where soil moisture was 30.5%, and 191mg/100g dry soil for the
control where soil moisture was 27% six weeks after the fire. The differences between the values of the MB in the control
and burned areas were significant only on two occasions, over all our experiment period. These occurred in both periods
when MB decreased significantly after it has reached its summit.
This suggests that MB renewal is faster in burned sites. Also, the saturation of burned soil surface (0-2 cm), where
the water content ranging from 46 to 54% in the respective period from mid-April to mid-May, was expressed in 2-10 cm
depth by optimal conditions of moisture content (27 and 23% respectively). The possible enrichment in leaching nutrients
and labile organic substances might be an additional cause having favored the above mentioned renewal. In all cases, it
seems that soil surface saturation and subsequent water repellent formation have positive effect on MB of the burned plot
at 2-10 cm depth and inhibiting effect on it at 0-2 cm depth.

CONCLUSIONS
Our experiment in the described circumstances of Aleppo pine ecosystem in Mediterranean zone shows an
immediate increase in soil MB. As a result, programmed burning of low intensity realized during early spring season could
be a management tool of soil improvement. However, fallowing a strong rainfall, an unexpected reduction in MB occurs in
soil surface due most probably to a temporary suppression of the aerobic microorganisms and possible substitution of
anaerobic microorganisms by aerobic ones. Accordingly, fire intensity and environmental and edaphic factors prevailing
both during and after burning determine deeply the tendency of soil biological processes. Nevertheless, changes in MB in
field studies cannot be related directly to any specific causative effect. Therefore, the effects of the factors that function in
burning should be investigated one by one with a more detailed approach.

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Ahmed Fahd Rachid

Table 2: Changes in Soil Microbial Biomass Carbon (Mg / 100g. Dry Soil) and Water Content (%) at 2-10 cm Depth
Date

MBC mg/100g. Dry Soil Water Content (%)

Control Burned

30 / 3
Fire date

93
(19)

Control

Burned

NS

111
(26)

10.41

13.84

129NS
ND
12.46
(6)
140
189NS
22.86
26.20
11 / 4
(40)
(23)
65
146*
19 / 4
20.33
26.61
(6.5)
(47)
59
86NS
02 / 5
20.4
34.6
(21)
(19)
191
216NS
26.88
30.51
16 / 5
(19)
(82)
**
158
209
30 / 5
13
22.5
(9)
(11)
137
140
13 / 6
10.12
17.10
(12)
(9)
NS
118
141
7.14
10.70
27 / 6
(14)
(10)
NS
119
145
25 / 7
3.7
7.28
(11)
(18)
5/8
5.5
7.5
ND
ND
NS
Each Value is the Average of Four Replicates. : Not-Significant Difference. *: Significant Difference At 95% of
Probability. **: Significant Difference At 99% of Probability. ( ): Standard Deviation. ND: No Determined
31 / 3

ND

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(Pinus halepensis) Ecosystem Around Montpellier City, South of France

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Ahmed Fahd Rachid

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Impact Factor (JCC): 4.3594

Index Copernicus Value (ICV): 3.0