Environ Geochem Health

https://doi.org/10.1007/s10653-019-00412-5 (0123456789().,-volV)
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89().,-volV)

ORIGINAL PAPER

The mechanisms of biochar interactions

with microorganisms in soil
Andrey V. Gorovtsov . Tatiana M. Minkina . Saglara S. Mandzhieva .
Leonid V. Perelomov . Gerhard Soja . Inna V. Zamulina . Vishnu D. Rajput .
Svetlana N. Sushkova . Dinesh Mohan . Jun Yao

Received: 27 March 2019 / Accepted: 30 August 2019

Ó Springer Nature B.V. 2019

Abstract Biochar, a carbonaceous material, is increas-
ingly used in the remediation of the anthropogenically
polluted soils and the restoration of their ecological
functions. However, the interaction mechanisms among
biochar, inorganic and organic soil properties and soil
biota are still not very clear. The effect of biochar on soil
microorganisms is very diverse. Several mechanisms of
these interactions were suggested. However, a well
acceptable mechanism of biochar effect on soil microor-
ganisms is still missing. Therefore, efforts were made to
examine and proposed a mechanism of the interactions
between biochar and microorganisms, as well as existing
problems of biochar impacts on main groups of soil
enzymes, the composition of the microbiota and the
detoxification (heavy metals) and degradation (poly-
cyclic aromatic hydrocarbons) of soil pollutants. The
data on the process of biochar colonization by microor-
ganisms and the effect of volatile pyrolysis products
released by biochar on the soil microbiota were analysed
in detail. The effects of biochar on the physico-chemical
properties of soils, the content of mineral nutrients and
the response of microbial communities to these changes
are also discussed. The information provided here may
contribute to the solution of the feasibility, effectiveness
and safety of the biochar questions to enhance the soil
fertility and to detoxify pollutants in soils.
Keywords Biochar
Bacteria
Remediation

Co-sorbents
Pollution
Introduction
Biochar is widely used in soil amendment for
improving physico-chemical properties and the

A. V. Gorovtsov (&)
T. M. Minkina
G. Soja

S. S. Mandzhieva
I. V. Zamulina
V. D. Rajput
IVET, University for Natural Resources and Life
S. N. Sushkova Sciences, 1190 Vienna, Austria
Southern Federal University, Rostov-on-Don 344006,
Russia D. Mohan
e-mail: gorovtsov@gmail.com School of Environmental Sciences, Jawaharlal Nehru
University, New Delhi 110067, India
L. V. Perelomov
Tula State Lev Tolstoy Pedagogical University, Tula, J. Yao
Russia China University of Geosciences, Beijing 100083, China

G. Soja
AIT Austrian Institute of Technology, ERT, 3430 Tulln,
Austria

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biological activities of agricultural soils. However,
biochar impact on the agricultural system components
is still not very clear. Extensive but often controversial
information about the biochar effect on soil properties
and the soil microbial activities is available in the
literature (Andrade et al. 2015; Ouyang et al. 2013).
To date, mechanisms of interaction between biochar,
soil organic matter and the soil biota are not well
explained (Nguyen and Lehmann 2009; Orr and
Ralebitso-Senior 2016).
Biochar impact on soil microorganisms is rather
diverse. Many interaction mechanisms have been
proposed in the literature (Palansooriya et al. 2019;
Zhu et al. 2017). The possible biochar–microorgan-
isms interaction mechanisms are discussed in this
review. Many of these mechanisms including toxicity
and emission of volatile organic compounds (VOCs)
act directly on soil microorganisms. The other mech-
anisms affect the soil microorganisms indirectly, by
changing the soil properties, regulating the nutrients
availability and modifying the enzymatic activities.
The above-mentioned interactions do not work sepa-
rately, and all of them affect each other to some extent.
Existing problems in biochar impact on soil prop-
erties, on microbiota composition and the detoxifica-
tion of pollutants are considered in the present review.
Biochar is widely used to enhance the polyaromatic
organic hydrocarbons (PAHs) degradation and to
immobilize and detoxify heavy metals. The available
information may contribute to the rationality, effec-
tiveness and safety of biochar use to enhance soil
fertility and to detoxify soil pollutants.
Biochar colonization by microorganisms
The colonization of various substrates by bacteria and
the formation of biofilms on their surface have been
studied for many decades (Leben 1969; Costerton
et al. 1987; Donlan 2002). Furthermore, no general
conclusions about the physical and chemical processes
underlying these interactions have been possible as
there are a number of different substrates and
microorganisms with different properties. From the
material properties viewpoint, the surface roughness,
its topography, the free energy, the surface charge and
hydrophobicity are the main parameters for the
process of microorganism attachment (Rummel et al.
2017). The majority of the researchers concluded that
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Environ Geochem Health
microorganisms attach faster to hydrophobic non-
polar surfaces versus hydrophilic ones (Donlan 2002).
A comparative study of the biofilm development on
hydrophobic and hydrophilic surfaces has shown that
differences were not observed after 167 days of the
experiment, and the major factors in biofilm develop-
ment kinetics included the roughness of the
surface and grazing of bacteria by protozoa (Pedersen
1990).
The interaction between microorganisms and sub-
strates leads to multiple adaptation approaches. For
example, microorganisms can form various surface
structures, such as flagella pili and fimbriae. This leads
to changes in hydrophobicity of cell walls and
membranes and their surface charge. The production
of extracellular polymeric substances can be also
altered (Rummel et al. 2017). All these mechanisms
may improve the cell adhesion to the inhabited
surfaces.
An important biochar property is its porous struc-
ture. Micro (\ 2 lm)-, meso (2–50 lm)- and macro
([ 50 lm)-pores that can act as habitats for bacteria
and fungi are distinguished in the biochar structures
(Downie et al. 2009). A successful use of the beneficial
microorganisms immobilized on the biochar is indi-
cated for the treatment of corn with Pseudomonas
putida (Sun et al. 2016a, b), soybean (Glycine max)
with Bradyrhizobium japonicum (Glodowska et al.
2017), and cucumber with Enterobacter cloacae (Hale
et al. 2015).
The biochar components from pyrolyzed sugarcane
and corncobs extracted by water suppressed the
growth of Leocobacter sp. and Bacillus aryabhattai
(Bueno et al. 2018). The same strains cultivated on the
solid medium in the presence of biochar formed
colonies, produced exopolysaccharides and formed
biofilms. Thus, the dominant substances and the
functional groups on biochar surface, available for
microorganisms and causing the specific reactions,
affect biochar colonization by microorganisms. The
low molecular weight hydrocarbons on biochar sur-
faces may be utilized by the soil microorganisms as a
carbon source (Farrell et al. 2013). They can also play
a role of the signal molecules stimulating or inhibiting
microbial activity and plant growth (Graber et al.
2010).
In spite of successful biochar usage as habitat for
microorganisms (Egamberdieva et al. 2018), the
problem of its colonization effectiveness by the
Environ Geochem Health
natural microbial soil community has not been fully
solved. In particular, it was shown that biochar added
to a loamy soil (Eutric Cambisol) underwent slow
colonization by microorganisms in an experiment
lasted for 3 years (Quilliam et al. 2013a, b).
A slow colonization of biochar may relate to its
particle size and the diffusion-limited gas exchange in
the centre of biochar particles. On the other hand, after
the biochar application to a sandy soil (Entisol) and to
a sandy loam (Spondasol) an active pore colonization
by microorganisms has been noted after 15 months of
interactions (Mukherjee et al. 2014). Thus, not only
biochar characteristics but also soil type can affect the
bacterial colonization speed. Probably biochar colo-
nization speed is also connected with ageing processes
causing the change in surface property. At the same
time, organic mineral complexes are formed on the
biochar surface as well as inside pores that may also
modify the adsorption properties and contribute to the
colonization of biochar particles with bacteria (Kumar
et al. 2018). It has been shown that biochar ageing
process in acidic Planosol soils proceeds faster versus
calcareous black soils (Rechberger et al. 2017).
Biochar is mineralized faster in the soils where it
was previously used, demonstrating an active role of
microorganisms in the process of biochar mineraliza-
tion (Budai et al. 2016).
In addition to chemical surface properties and the
biochar particle size, the pore size distribution is also
an important factor for the colonization by microor-
ganisms. In the case of larger pores, the time of biochar
particle colonization increased, but large pores did not
protect bacteria from being grazed by soil fauna as
explained by Noyce et al. (2016).
The contact time affected microbial consortia
composition occupying the porous space (Quilliam
et al. 2012). Fresh biochars release organic substances
after their application to soil. This supports the growth
of the bacteria that are able to utilize these substrates
and leads to the biochar colonization (Dutta et al.
2017; Sun et al. 2015). When this source is depleted,
the communities of the porous spaces alter in the
course of microbial succession (Prayogo et al. 2014).
Genetic sequencing of microbes in biochar particles
incubated in the soil for 4 years has shown that inside
and in the direct proximity of biochar particles, a
decrease in the bacterial biodiversity and an increase
in the eukaryotic diversity have developed (Noyce
et al. 2016). The number of Acidobacteria,
Planctomycetes and b-Proteobacteria decreased, and
with respect to the eukaryotes, an increased number of
Aveolata protists were observed (Noyce et al. 2016).
Jin (2010) showed the modification of the fungal
community when biochar was applied. The biodiver-
sity shifted from the predominance of representatives
of the Basidiomycota and Ascomycota divisions in
untreated soils to the dominance of fungi from
Zygomycota, Glomeromycota and Neocalli-
mastigomycota divisions after the application of
biochar.
Thus, the process of biochar colonization by
microorganisms mainly depends on biochar composi-
tion. Biochar composition is mainly dependent on
biomass feedstock, residence time, pyrolysis temper-
ature and reactor type (Mohan et al. 2006). Other
important factors included soil properties, the abun-
dance and composition of the consortia of pre-existing
microorganisms in the soil and the biochar–soil
contact time (Jin 2010; Quilliam et al. 2012; Noyce
et al. 2016).
The process of biochar ageing can significantly
alter its properties and lead to colonization by
microorganisms (Mukherjee et al. 2014). It has been
shown that the addition of aged biochar increased
microbial activity, while the opposite was observed for
fresh biochar (Wang et al. 2016a, b). These findings
suggest that soil bacteria colonize aged biochar better
than fresh one. The most evident reason is in the toxic
effects of freshly prepared biochar due to high content
of pyrolysis products.
Substances emitted into the soil by biochar
and their effect on soil microorganisms
During biochar production, significant amounts of
new organic substances are formed which may
substantially influence soil properties and the inhab-
iting microorganisms (Spokas et al. 2011; Sun et al.
2016a, b). Therefore, the exact composition of fresh
biochar determines the changes in the microbial
communities in the initial stage after its addition to
soil. In the following section, the nature of substances
emitted by fresh biochar and the effects on microbial
communities are discussed.
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Volatile organic compounds (VOCs)
Volatile organic compounds (VOCs) are the most
widely studied groups of organic compounds formed
in the biomass pyrolysis process (Dunnigan et al.
2018; Li et al. 2019; Zheng et al. 2019). It has been
shown that various biochar may contain more than 140
VOCs (Graber et al. 2010; Ghidotti et al. 2016). At the
same time, their contents do not primarily depend on
the initial raw material but on the biochar production
conditions (Spokas et al. 2011).
Organic compounds including methanol, acetone,
benzene, methylethylketone, toluene, methyl acetate,
propionaldehyde and benzo[a]pyrene were deter-
mined in different biochars (Spokas et al. 2011;
Ghidotti et al. 2016). Biochars also contain ethylene,
propylene, ethane, propane, butane, acetylene, pen-
tane and hexane (Sánchez et al. 2009). Various
aliphatic and aromatic acids and phenols are often
present (Buss et al. 2015; Buss and Mašek 2016;
Ghidotti et al. 2016; Spokas et al. 2011). The presence
of toxic effects for plants and microorganisms has
been shown for many of these substances (Dutta et al.
2017). The data of the impact of VOCs emitted from
biochar on microorganisms are presented in Table 1.
In all cases, the presence of a complex mixture of
volatile compounds had been observed. The detailed
composition about these compounds is described in
the cited articles.
Both positive and negative effects of biochar on
various microbiological processes in the soil were
reported (Spokas et al. 2010; Wang et al. 2015). In
some studies, a decrease in nitrification level was
recorded during biochar use (Spokas et al. 2010; Wang
et al. 2015), while other studies showed an opposite
trend (Wang et al. 2018a, b, c; Abujabhah et al. 2018).
Thus, it is very difficult to establish any single effect of
biochar on the microbial communities for the emission
of VOCs.
A strong toxic effect of VOCs has been mentioned
in the majority of studies. On the other hand, the
consideration of the dependence of these effects on the
initial raw material and the method of biochar
production may mitigate or prevent negative aspects
of the biochar applications (Buss et al. 2015; Chen
et al. 2013).
Thus, before wide use of biochar in agricultural
practices, new methods for potential toxicity assess-
ment should be developed. This is necessary because
the biochar composition is not limited by

Table 1 The impact of biochar volatile organic compounds on the microorganisms

Biochar precursor Pyrolysis Volatile organic compounds and their impact on the References
temperature microorganisms

Steam-activated charcoal and 12 350–550 °C Inhibited nitrification which is associated with the action of Spokas et al.
different biochars ethylene (2010)
Corn stems, the substrate for 450 °C The high content of phenolic compounds decreased the number of Sun et al.
growing fungus and rice straw Bacillus mucilaginosus (2015)
Sawdust 400, 500, The use of bioluminescent strains Pseudomonas putida and Ps. Knox et al.
600 °C fluorescens has revealed a higher biochar toxicity obtained at (2018)
500 °C in comparison with 400 °C or 600 °C that is linked with
the presence of some toxic compounds
Rice straw 400–500 °C The changes in the microbial communities of the compost were Sun et al.
noted, while increasing biodiversity and expanding the range of (2016a, b)
substrates used
Rice husk, sawdust, Acorus 650 °C Ps. aeruginosa strains were used as a model. Slight toxic effects of Wang, et al.
calamus plants biochar from sawdust and rice husk and a high toxicity of biochar (2016a, b)
from Acorus calamus due to the presence of low molecular
weight aromatic compounds has been shown
Cellulose, lignin and pine wood 300, 400, It has been shown that the toxic effect of biochar on cyanobacteria Smith et al.
500 °C Synechococcus is associated with the formation of phenolic (2016)
compounds in the case of wood and lignin, and organic acids in
the case of cellulose. In this case, the toxicity decreased with
increasing temperature of pyrolysis

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comparatively easily biodegradable volatile com-
pounds, but also contains more stable organic mole-
cules (Freddo et al. 2012).
Polyaromatic hydrocarbons (PAHs)
Polycyclic aromatic hydrocarbons (PAHs) are formed
at incomplete fuel combustion and act as the leading
organic pollutants due to their carcinogenetic, muta-
genic and teratogenic properties (White and Claxton
2004). PAHs, exhibiting carcinogenic, mutagenic and
toxic properties in relation towards living organisms,
are included in the list of permanent monitoring
substances by the European Union and the USA
Environmental Protection Agency (EPA) (European
Commission JRC 66955 2011; U.S. EPA 2003; IARC
1983a, b).
PAHs may be transferred in the atmosphere for a
great distance, even reaching polar regions (Fernandez
et al. 2002; IARC 1983a, b). PAH formation inevitably
occurs in the pyrolysis process to obtain biochar and
causes concern regarding the safety aspect of wide-
spread biochar use in agricultural soils (Hale et al.
2012). According to IBI recommendations, the upper
limit of PAHs content in biochar ranged from 6 to
300 mg/kg (IBI 2015). The more prudent EBC
guidelines (European Biochar Certificate) classified
two different biochar qualities with \ 4 and \ 12 mg
PAH/kg (EBC 2012).
Biochar applications may impact the soil due to the
PAHs that are produced during the pyrolysis (Kuś-
mierz et al. 2016; Quilliam et al. 2013b). Biochar may
contain significant quantities of PAHs depending on
precursor. A 64.65 mg/kg of total PAHs was reported
in rice husk biochar, while 9.56 mg/kg was deter-
mined in wood biochar. The dominating PAH was
naphthalene (Quilliam et al. 2013b).
When producing biochar from sewage sludge, the
naphthalene content was increased; however, the
content of PAHs with a large number of aromatic
rings decreased compared to the initial raw material
(Zielinska and Oleszczuk 2015). Nevertheless, due to
high sorption ability, the PAHs formed during biochar
production may remain sorbed to the biochar without
entering the soil solution, thereby limiting the poten-
tial ecotoxicological risks (Hale et al. 2012). In this
way, it is reasonable to assess the potentially bioavail-
able PAHs content in biochar but not their total
content. Mass studies (more than 60 various biochar)
have shown that the bioavailiable PAH content in
biochar lies in the range from 0.2 to 2 ng/l, which is a
much lower level than the maximum permissible
concentration (Cornelissen and Hale 2017).
On the other hand, the biochar may sorb PAHs,
thereby decreasing their bioavailability. Regarding
this issue, the data in the literature are contradictory.
Particularly, it has been shown that the biochar
application to soil contaminated by phenanthrene
results in PAHs content that was higher than that
without biochar after 3 years (1.95 mg/kg vs.
1.13 mg/kg) (Quilliam et al. 2013b). In another work,
the application of biochar to the soil mixed with
wastewater sediments was shown to lead to the
decrease in PAH dissolved content, especially, in 3-,
5- and 6-ring PAHs (Stefaniuk et al. 2018). The
increase in the total and decrease in bioavailable PAH
content were also observed when using bamboo
biochar on the rice field soil (Ni et al. 2018). Thus, a
high sorption ability of biochar blocks the PAH
bioavailability, which prevents their inclusion in the
food chain and decreases the risk to ecosystems.
However, at the same time, such immobilization may
slow the PAH microbial degradation process in the
soil.
Less information is available on the impact of
PAHs present in biochar on microorganisms. This is
due to the fact that, in contrast to VOCs, it is
practically impossible to separate the impact of PAHs
sorbed on biochar and the impact of biochar itself.
This is due to high content of aromatic C embedded
into the structure of biochar (Keiluweit et al. 2010).
Nevertheless, the latest work mentioned that a number
of bacteria negatively correlated with the aromatic
carbon content in biochars applied to the soil and, on
the contrary, positively correlated with the aliphatic
carbon content (Zhang et al. 2018a, b, c). The toxic
impact of biochar on the bacteria Vibrio fisheri has
been shown to depend on the PAH content, while PAH
removal from biochar by the use of thermo-treatment
decreased sorbent toxicity (Koltowski and Oleszczuk
2015).
The negative impact of PAHs present in biochar is
not limited to direct toxic effect on the soil bacteria
(Gorbov et al. 2015). PAHs in the soil are able to
increase the antibiotic resistance of the soil bacteria
due to the horizontal transfer of PAH degradation
genes and ARGs on the same plasmids (Gorovtsov
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et al. 2018). It has special importance in the case of
biochar, as its pores may act as microenvironments,
where the most resistant bacterial communities may be
formed (Zechner et al. 2000). This can be assumed
from the facts that horizontal gene transfer is easier
when microbial cells are close to each other (Zechner
et al. 2000), and from high concentration of PAHs in
biochar pore space that could promote co-selection of
the ARGs. This process is promoted by biochar use in
the form of mixtures with compost or manure (Yuan
et al. 2017), which themselves are the sources of
resistance genes.
Mineral nutrient elements in biochar composition
and their impact on microorganisms
Biochar precursors including wood, straw, litter,
composts and agricultural residues always contain a
large amount of mineral elements, which remain in the
biochar after pyrolysis. Carbon begins to transfer into
gaseous state at 100 °C, nitrogen at 200 °C, sulphur at
375 °C and phosphorus and potassium at 700–800 °C
(Deluca et al. 2015). These nutrients can then slowly
diffuse into the soil and are used by microorganisms
(Zhu et al. 2017) and plants (Amini et al. 2015). It has
been shown that a significant proportion of potassium
and phosphorus contained in biochar granules passes
into the soil solution (Limwikran et al. 2018). At the
same time, the nitrogen in the low-temperature peanut
hull and pine chip biochar does not mineralize, which
is why it may not be regarded as a source of plant
mineral nutrition (Harris et al. 2013). Mineral nutrient
elements may sorb on biochar granules due to the
electrostatic attraction. (Potassium, ammonium and
magnesium cations interact with the negatively
charged hydroxyl and carboxyl groups.) The forma-
tion of hydrogen bonds between the dihydrogen
phosphate anions and carboxyl groups in the undisso-
ciated state is also possible (Chen et al. 2014a, b). The
stability of these bonds is directly connected to the pH
level of the soil solution, and the sorption and
desorption process of the nutrient elements may occur
at local changes. The treatment of mineral elements
with wood ash had an evident positive effect on soil
microbial communities (Mahmood et al. 2003). The
analogous effect may be expected in the case of
biochar rich in mineral ash element application. This is
indirectly confirmed by the data (Fox et al. 2014),
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Environ Geochem Health
which have shown the activity increase in phosphate-
and sulphate-mobilizing bacteria during biochar appli-
cation to the soil.
On the other hand, there are numerous studies
demonstrating that the soil’s ability to keep nutrients
depend on the pyrolysis temperature (Cheng et al.
2018), the gas media applied (Tan et al. 2018), and on
biochar modification. Thus, the nutrients accumula-
tion in soil during the biochar composting included the
formation of active layer of the organic substances on
their surface (Hagemann et al. 2017; Joseph et al.
2018). It is very difficult to separate the impact of the
inherent biochar mineral substances on microorgan-
isms, and the nutrients entered the biochar during the
sorption process from the soil solution. Obviously, as
in cases with VOC, the inherent mineral elements are
responsible for the short-term effects after the appli-
cation and the long-term effect provided by biochar
interaction with mineral nutrition element uptake from
the soil. The biochar capacity to concentrate nutrition
elements on its surface creates favourable conditions
for colonization by microorganisms, especially in poor
soils. Besides, the availability of mineral elements and
the enhanced microbiological activity may be con-
nected with the cation exchange capacity, increasing
with biochar application (Andrade et al. 2015).
Biochar and microelement interactions
A more variable situation may develop with regard to
the availability of trace elements. On the one hand,
there are data showing that biochar may act as the
source of boron and molybdenum, intensifying nitro-
gen fixation (Lehmann et al. 2011). On the other hand,
the biochar capacity to effectively sorb the cations at
definite conditions may lead to negative consequences
(Fidel et al. 2018; Gao et al. 2019). In particular, iron
deficiency has been revealed in plants grown on
carbonate soils with biochar use. In this case, the
active formation of hydroxide iron has been shown on
biochar grains (III) (Sorrenti et al. 2016). It should be
noted that iron accessibility might be a quite signif-
icant factor for the microbial communities. One of the
key mechanisms of bacterial antagonism against the
phytopathogenic fungi is the siderophores synthesis,
which limits iron accessibility for the fungus (Chin-
cholkar et al. 2000). This mechanism is the basis of the
antifungal activity of pseudomonads (Kotasthane et al.
Environ Geochem Health
2017), rhizobia (Gupta et al. 2018) and some other
kinds of bacteria (Maleki et al. 2018). The high
capacity of biochar to limit iron accessibility should be
the basis of suppressing the root rot mentioned in
many research articles (Jaiswal et al. 2017; Rogovska
et al. 2017).
However, the iron deficiency may exert a suppres-
sive action on beneficial fungi, e.g. mycorrhizae-
forming ones by disturbing the P/Fe ratio (Paymaneh
et al. 2018; Wang et al. 2008). In some investigations,
it has been shown that mycorrhizae-forming fungi
need a considerable amount of iron (Haselwandter
1995; Winkelmann 2017). There are also some data
showing that biochar application may have lower
positive effects due to the interaction of mycorrhizae-
forming fungi with plants (Liu et al. 2017a, b, b).
Nevertheless, the majority of works mention the
stimulatory effect of biochar (Hammer et al. 2015;
Mickan et al. 2016) that may be linked to the diversity
of the mechanisms underlying its impact on the soil,
fungi and plants (Koide 2017). Biochar application
may affect the dynamics of other microelements. The
biochar application into the soil decreased the yield
and concentrations of micronutrients (Cu, Fe, Mn and
Zn) in the tissues of mustard (Sinapis alba L.), barley
(Hordeum vulgare L.) and red clover (Trifolium
pretense L.), while an increase was noticed in Mo
concentration) (Kloss et al. 2014). In other cases, the
decrease in Zn bioaccessibility was observed in rye
(Secale cereale) (Wu et al. 2018a, b). On the other
hand, directly opposite data have been obtained for
rice, where the Zn bioaccessibility increased (Jatav
et al. 2018).
Summarizing the above-mentioned points, the use
of biochars on soils with high pH, carbonate soils and
those with low microelement content may induce no
significant or even unwanted effects. Due to the high
sorption capacity the biochar should be used with
mineral or organic fertilization or should be utilized a
post-pyrolysis biochar modification (composting, fer-
mentation, microfertilizer combination) (Kloss et al.
2014).
Impact on soil properties and their microbial
communities
A considerable part of the positive effects of biochar,
allowing its use in agricultural practice as an
ameliorant, is linked with the improvement of physical
and chemical properties (density, microaggregate
stability, pH) and water–air soil regime (Blanco-
Canqui 2017; Burrell et al. 2016; Ding et al. 2016; Gul
et al. 2015). All of these effects produce a direct
impact not only on the plant root system but also on the
rhizosphere microbial communities (Igalavithana
et al. 2017; Liang et al. 2014; Liu et al. 2017a, b; Ni
et al. 2017). Biochar application decreases the bulk
density and increases the moisture-holding soil capac-
ity (Andrenelli et al. 2016). Simultaneously, the
biochar application affects both nitrifying and deni-
trifying bacteria. The moisture content in the soil has
been shown to affect the final N2O emission signif-
icantly. When biochar was introduced into the soil
with moisture less than 70% of full capacity, an
increase in N2O emission was observed due to biochar
stimulation of nitrifying bacteria. At more than 70%
moisture content, anaerobic bacteria reducing N2O to
N2 were observed (Liu et al. 2017a, b). Obviously,
although biochar application reduces the soil bulk
density, this does not lead to a significant increase in
soil aeration and the disappearance of anaerobiosis
zones at an appropriate humidity level (Blanco-
Canqui 2017). In this case, the N2O emission lowering
may be explained by nitrate immobilization and the
decrease in accessibility for denitrifying bacteria
(Case et al. 2012).
Biochar impact on pore size in the soil and the
amount of moisture accessible to plants remains the
object of discussion (Ahmad et al. 2014; Ameur et al.
2018; Andrade et al. 2015; Beesley et al. 2010;
Blanco-Canqui 2017; Gul et al. 2015; Hagemann et al.
2017; Hardie et al. 2014; Jien and Wang 2013;
Lehmann et al. 2011; Liu et al. 2017a, b, 2018;
Shaaban et al. 2018). Hardie et al. (2014) reported that
a direct biochar impact on soil moisture was not
observed. At the same time, the biochar impact on the
moisture-holding capacity and an accessible moisture
content depended on the granulometric soil composi-
tion. It has been shown that the application of biochar
to light loamy soil (sandy loam) leads to an increase in
water-holding capacity (Dan et al. 2015); on a middle
loamy soils (silt loam), there were no changes (Aller
et al. 2017), but a decrease in these indices was
observed in a heavy loamy soil (clay loam) (Aller et al.
2017).
Nevertheless, biochar grain capacity to hold water
obviously produces a favourable effect on the
123

microbial communities even if this water remains
slightly accessible for the plants (Liang et al. 2014).
Particularly, soil microbial communities, in which
biochar is added, are restored much more quickly after
the drought period (Liang et al. 2014). It has been
stated that biochar application increases the metabolic
activity of the soil microbial communities in the
drought conditions; the ‘‘aged’’ biochar happens to be
more effective than fresh one (Paetsch et al. 2018).
Biochar impact on soil structure and the microbial
communities’ reaction to these changes
A number of studies were carried out about the biochar
impact on the soil aggregate stability (Gul et al. 2015;
Ibrahim et al. 2013; Jien and Wang 2013; Ouyang et al.
2013; Soinne et al. 2014; Khademalrasoul et al. 2014;
Zheng et al. 2018). Since these structural parts of the
soil are substrates for microbial biofilm development,
any change in their formation process and their
existence in the soil media produce a significant
impact on the microbiological processes.
In many studies, there was a positive effect on
microaggregate formation (Ibrahim et al. 2013;
Demisie et al. 2014; Zhang et al. 2017a, b), although
in some cases there were no differences with the
control (Rahman et al. 2017; Zhang et al. 2018a, b, c)
(Table 2).
It is possible to consider that the formation of
aggregates occurs around biochar grains. The mycelia
of microorganisms belonging to Actinobacteria filum
are actively involved in this process (Luo et al. 2018;
Zheng et al. 2018). Hyphae of fungi and actinobacteria
may contribute to the increase in cohesion between the
soil particles and the structural aggregates of various
sizes with the biochar grain surface, leading to the
aggregation increase (Zheng et al. 2018). The sapro-
trophic basidiomycete fungal hyphae produce
exopolysaccharide with adhesive properties and play
an important role in aggregation of soil clay particles
(Caesar-TonThat et al. 2010). The microscopic anal-
ysis confirmed the active role of filamentous acti-
nobacteria (actinomycetes) to form soil aggregates. It
is well known that hyphal networks bind soil particles
and form aggregates (Foster 1988; Frey 2015). Soil
microorganisms could connect soil mineral particles
or aggregates and stabilize microaggregates (Tiessen
and Stewart 1988). The stability of microaggregates
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Environ Geochem Health
can be further enhanced because mycorrhizal fungi
Glomus fasciculatum and Acaulospora laevis can
promote the growth of Streptomyces actinobacteria
(Secilia and Bagyaraj, 1987). It was reported that the
arbuscular mycorrhizal fungi and actinobacteria col-
onize the surface of biochar granules (Thies and Rillig
2009). A recent study also showed that the application
of biochar promotes fungal and actinobacterial growth
(Luo et al. 2018). The pore space of biochar could be a
harbour for the hyphae of AM fungi. Inside a biochar
pore space, microorganisms exist in a protected
environment from other soil organisms and could be
a source of inoculum (Koide 2017). Therefore, the
biochar particles could promote soil aggregation with
the support of soil microorganisms, and this mecha-
nism is partially explained in Table 2.
Impact on soil pH and microbial communities
Biochar application is always followed by a more or
less substantial soil pH increase (Blanco-Canqui 2017;
Burrell et al. 2016; Fidel et al. 2018). These changes
may affect a number of agrochemical and biological
soil properties including microbial communities’
composition, soil enzymes and mineral feeding ele-
ment availability (Gao et al. 2019; Krause et al. 2018;
Palansooriya et al. 2019; Zhang et al. 2018a, b, c).
Soil pH increase at biochar application is due to
various reasons. First, the biochar itself has an alkaline
reaction; at the same time, biochar obtained from
wood, as a rule, has a higher pH in comparison with
that obtained from other sources (Gul et al. 2015).
In addition, the pyrolysis temperature has a sub-
stantial effect on biochar pH. It was shown that an
increase in Conocarpus pyrolysis temperature further
increases the biochar pH from 6.69 at 200 °C to 9.67 at
400 °C and 12.38 at 800 °C. This increase was
directly connected with the increase in ash content in
the biochar (Al-Wabel et al. 2013). Secondly, the
biochar has sufficient amount of negatively charged
functional groups that may link the hydrogen ions,
leading to a pH change toward the alkaline side
(Brewer and Brown 2012).
In this case, substantial changes in microbial
communities following biochar application into the
soil due to pH change were observed in many studies
(Krause et al. 2018; Zhang et al. 2018a, b, c; Palan-
sooriya et al. 2019). Thus, there is a substantial impact
Environ Geochem Health

Table 2 The impact of biochar on the soil structure and microbiological activity
Biochar precursor Pyrolysis Soil type and particle Impact on soil aggregation and microbial References
temperature distribution communities
(°C)

Conocarpus wood 400 Typic Torriorthents light loamy The application of 1.5 and 2% biochar increased Ibrahim et al.
soil water-resistant aggregates of 1–2 mm and 0.5% (2013)
biochar increased in the share of water-resistant
aggregates of 1–0.5 mm and 0.5–0.25 mm
Leucaena 700 Typic Paleudults, clay loamy soil 63 days of incubation of 5% biochar significantly Jien and Wang
leucocephala (Lam. increases the stability of the aggregates. Increase (2013)
De Wit) wood in the microbial biomass carbon content at the
same time
Manure 500 Farmlands’ soils, clay loam, The addition of 2% biochar significantly increased Ouyang et al.
loamy soil the share of macro-aggregates in both types of (2013)
soil. Light loam showed a more pronounced
response to the addition of biochar
Oak wood Quercus 600 Plinthustults Clay loamy soil A significant increase in the water resistance of the Demisie et al.
phillyraeoides and aggregates at the application of 0.5 and 1% (2014)
bamboo biochar. The increase in microbial biomass and
Phyllostachy edulis activity of beta-glucosidase and dehydrogenase
Fir (Picea abies wood 550–600 Endogleyic Stagnosol clay loamy An increase in the share of water-resistant Soinne et al.
and pine (Pinus soil aggregates was observed, depending on the (2014)
sylvestris) wood Cutanic Vertic Luvisol clay incubation mode: clay soil better aggregated in
the presence of biochar when being dried, and
loam in a wet state
Birch tree wood 500 Corn field according to no-till An increase in the stability of large aggregates of Khademalrasoul
(Betula) technology Light loamy soil 4–8 mm and 8–16 mm, their strength index and et al. (2014)
a decrease in the erosion of clay particles were
noticed after applying high doses of biochar
(0–10 kg m-2)
Wheat straw 350–550 Earth cumuli-Orthic Anthrosols, The use of biochar in a dose of 16 t/ha led to an Zhang, et al.
Clay loamy soil increase in the number of macro-aggregates (2017a, b)
([ 2 mm) by 106%. Increases noted in
microbial biomass carbon, nitrogen and urease
activity
Corn stems ND Vertisol, clay loamy soil In the variant with biochar, there were no Rahman et al.
differences from the control on aggregation and (2017)
microbiological activity
Corn stems 350, 500 Fluvisol clay Increasing the stability of microaggregates due to Zheng et al.
the interaction of biochar with clay minerals, an (2018)
increase in microbial biomass and a decrease in
carbon mineralization. Increase in the share of
taxa of bacteria stabilizing microaggregates
(Actinobacteria, Acidobacteria.)
Corn stems 550 Silandic, Vitric Andosols, loamy In the variant with biochar, there were no Sarker et al.
soil; Xerofluvent, loamy soil; differences from control in the aggregates (2018)
Typic Ustifluvent sandy loamy formation
soil

on the communities’ composition of nitrifying
microorganisms. Biochar application led to an
increase in the representatives’ share of Nitrosomonas
and Nitrospira in the nitrifying bacteria community
and mostly affected the ratio of ammonium-oxidizing
bacteria and archaea, which is due to an ammonium
availability increase (Zhang et al. 2018a, b, c). The
number of bacteria increased, restoring N2O at biochar
application due to pH increase. It has been shown that
there are similar effects at the soil liming (Krause et al.
2018). A substantial impact of the initial soil pH where
biochar is applied on the following microbiological
processes development has also been observed
(Igalavithana et al. 2017; Zhang et al. 2019). Thus, a
more intense biochar impact on CO2 and N2O
emission is suggested at application into acidic soil

123

in comparison with alkaline soils (Wu et al. 2018a, b).
In another recent study, it was detected the biochar
application into acidic soils and the pH increase
associated with it leads to changing the taxonomic
composition of microbial communities. The share of
copiotrophs from the phylum Bacteroidetes and
Gemmatimonadetes increases with the share of olig-
otrophs composition of microbial communities, and
the share of Acidobacteria phylum decreases, which
leads to CO2 emission intensification. In alkaline soils,
an opposite situation is observed (Sheng and Zhu
2018).
Thus, we may conclude that a substantial pH
variation caused by biochar application is the one of
the leading mechanisms of its effect upon microbial
soil communities. The intensity of this effect is
stronger in acidic soils with a low organic substance
content; therefore, there is low buffering. In the
majority of cases, the acidic soils’ alkalinization by
biochar produces a positive effect due to the alu-
minium toxicity decrease and the increase in available
nutrients. A negative effect may be organic substance
mineralization intensification and carbon dioxide
emission into the atmosphere.
Impact on soil enzymatic activities
A diverse biochar impact on the soil properties as well
as on its activity and the number of microbial
communities directly affects the soil enzymatic activ-
ity. Among possible mechanisms of a direct biochar
interaction with enzymes, one may observe the
enzymes or their substrates adsorption on biochar
(Bandara et al. 2017; Wang et al. 2018a, b, c; Yang
et al. 2016). Biochar impact on soil solution pH is
great, which may change its values closer to or farther
from the optimal one for a specific enzyme. The
influence through the sorption of trace elements,
which serve as the cofactors for many enzymes, is also
possible (Pukalchik et al. 2018; Lehmann et al. 2011).
Finally, the biochar participation in electron transport
as well as the presence of free radicals on its surface
(Fang et al. 2014) may present interest in the context of
interactions with enzymes of the oxidoreductase class.
The indirect impact of biochar on soil enzymatic
activity is due to its capacity to stimulate specific
microorganisms groups, to interact with plant root
systems that may affect the expression level and
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Environ Geochem Health
secretion of some enzymes (Glodowska et al. 2017;
Hale et al. 2015; Sun et al. 2016a, b). Among the six
existing classes of enzymes, only two are widely
known in soil studies. Enzymes of the oxidoreductase
class participate in redox reactions; they are linked
with the use of different substrates by microorganisms
and responsible for an important part of the humifi-
cation process (Sun et al. 2016a, b). To this class,
enzymes such as polyphenoloxidase, nitroreductase,
peroxidase and catalase belong, along with a wide
spectrum of dehydrogenases (Bandara et al. 2017;
Rizhiya et al. 2017; Teutscherova et al. 2018).
Enzymes of the hydrolase class participate in the
decomposition of the organic substance entering the
soil. Hydrolases such as urease, beta-glucosidase and
various kinds of protease, xylanase, cellulase, pecti-
nase, phosphatase and others have been studied
(Pukalchik et al. 2018; Liu et al. 2018; Yang et al.
2016).
It can be seen from the given data (Table 3) that
biochar has a controversial effect on the soil enzymatic
activity. At the same time, some regularities may be
observed which allow conclusions to be drawn about
separate mechanisms of biochar impact on the enzy-
matic activity.
Biochar impact on hydrolase activity
In most of the research articles, a positive impact of
biochar was confirmed on the urease activity that
should be specified by the pH of soil solution increase
(Blanco-Canqui, 2017; Burrell et al. 2016; Fidel et al.
2018). Thus, in the work of Pukalchik et al. (2018), the
activity of enzymes with a high pH (* 7.0) increased
urease and decreased the activity of acid phosphatase.
A more obvious example of pH impact is shown by
Liu et al. (2018). In this work, biochar treated with
aqueous HCl and subsequent washing was used
alongside untreated biochar. The untreated biochar
stimulated the urease activity more intensively and the
treated one stimulated acid phosphatase activity due to
the removal of ashy elements by the acidic treatment
(Liu et al. 2018). Nevertheless, the same study showed
that the impact of biochar on the enzymes is not only
due to its action on the medium reaction, as biochar
treated with acid produced a stimulatory effect upon
urease activity (Burrell et al. 2016). A positive effect
of biochar application may be due to the enzyme-
Environ Geochem Health

Table 3 Biochar impact on oxidoreductases and hydrolases activity

Biochar precursor Pyrolysis Impact on oxidoreductases activities Impact on hydrolases activities References
temperature
(°C)

Bamboo, Biochar 500, 750 Intense increase in catalase activity Intense increase of urease activity Yang et al.
from rice straw (2016)
Gliricidia sepium 300, 700 Decreased the activity of catalase, ND* Bandara et al.
dehydrogenases and polyphenol oxidase (2017)
and with addition of fungi and bacteria
biomass significant increase in the activity
of these enzymes
Gliricidia sepium 300, 500, 700 With the addition of 1, 2.5 and 5% biochar, ND Vithanage
a decrease in the activity of polyphenol et al. (2018)
oxidase, an increase in the activity of
catalase (up to 34%), and a pronounced
increase in the activity of dehydrogenases
(up to 3.5 times) were observed
At the same time, for biochars with
increased pyrolysis temperature, the
positive effect decreased and an inhibitory
effect was observed
Birch, aspen 550 The catalase activity increase by 1.3 times, No intense impact on urease activity Rizhiya et al.
also significant increases observed in the (2017)
activity of polyphenol oxidase and
peroxidase
Acer rubrum and 200, 300, 450, The wood species dependent effects were ND Gibson et al.
Pinus baksiana 600 observed, maximum activity of (2018)
peroxidase and polyphenol oxidase was at
a pyrolysis temperature of 300 °C
Coco shell before 800 The activity of dehydrogenases dose- The increase in urease activity was stronger Liu et al.
and after treatment dependently decreased after 7 days and with untreated biochar, and acid (2018)
by HCL increased after 63 days of incubation phosphatase with treated one. Invertase
activity was increased in both cases
Oak wood 600 Dose-dependent dehydrogenase activity Dose-dependent decrease in beta- Teutscherova
increase glucosidase, beta-glucosaminidase and et al. (2018)
acid phosphatase activity, increase in
urease activity
Wood 700–900 Dehydrogenase activity increase Reduced acid phosphatase activity, Pukalchik
increased urease and beta-glucosidase et al. (2018)
activity
Pine needles, and 450 °C for ND Alkaline phosphatase activity increase, Sa nchez-
coffee ground 2 h then beta-glucosidase activity decrease, no Hernandez
300 °C for effect on aryl sulfatase (2018)
3h In the presence of earthworms, a positive
effect of the biochar on the enzyme
activity was observed
Barley straw 600 ND An increase in the activity of acid Wang et al.
phosphatase and urease was observed (2018a, b, c)
with the use of biochar, and was enhanced
by its joint application with vermicompost
in soils contaminated with cadmium
Wood 450 ND There was a change in the activity of Ameur et al.
exoglycanase, beta-glucosidase, chitinase (2018)
and leucine-aminopeptidase when the
biochar activated with organic acids
mixed with composts was added to the
soil, depending on the incubation time: an
increase in activity after 1–7 months and
a decrease in 22 months

ND not determined

123

inhibiting substance sorption that is indirectly con-
firmed by the data on biochar sorption, using both
natural (Clough et al. 2013) and artificially synthe-
sized nitrification inhibitors (Keiblinger et al. 2018).
For some enzymes, namely beta-glucosidase and
beta-glucosaminidase (chitinase), the opposite is true
(Pokharel et al. 2018). A decrease in enzymes activity
was observed in the response to biochar application
(Sanchez-Hernandez 2018; Teutscherova et al. 2018).
An inhibitory action has been observed when utilizing
biochar at 300 and 550 °C (Pokharel et al. 2018). This
may provide evidence in such conditions of inhibiting
substances. Among the potential inhibiting substances
that can reduce beta-glucosidase activity are phenol
compounds, i.e. polyphenols (Lammirato et al. 2011).
Besides, the most important mechanism of biochar
impact on the soil’s enzymatic activity is the direct
adsorption of both enzymes themselves and their
substrates on biochar. Thus, it has been shown that the
biochar absorbed up to 99% of beta-glucosidase;
hydrolysis activity was 30% lower (Lammirato et al.
2011). The same mechanism may be the basis of the
protective action of biochar against some plant diseases.
It has been stated that biochar sorbs and inactivates
enzymes emitted by phytopathogenic fungi, which
destroy plant cell walls (Jaiswal et al. 2018).
Thus, taking into account, the significant amount of
data about biochar having an inhibitory effect, as well
as the stimulatory effect of biochar on various soil
exoenzymes, we suggest a complicated character of
adsorption process effects upon enzymatic activity. At
the same time, granulometric soil composition may
play a significant role. In particular, it has been shown
that the beta-glucosidase activity decrease at biochar
introduction into soils with a lighter granulometric
composition was expressed more clearly than in soils
with the heavier granulometric composition (Günal
et al. 2018). This result may be specified by enzyme
adsorption by organic mineral soil complexes that
preserve their activity. In addition, various interac-
tions are possible between biochar and clay loamy soil
minerals, which can affect its interaction with
enzymes.
Biochar impact on oxidoreductase activity
Oxidoreductase class enzymes comprise both extra-
cellular and intracellular enzymes catalysing redox
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Environ Geochem Health
reactions (Minnikova et al. 2017). A stimulating
effects as well as inhibitory activity have been shown
on catalase, polyphenoloxidase, various peroxidases
and dehydrogenases when applying biochar in the
soils.
A number of biochar effect mechanisms upon
oxidoreductase activity coincide with those described
for hydrolases, including pH change, and sorption of
the inhibitory substances, substrates and enzymes
themselves on biochar. In particular, it has been shown
that biochar artificial ageing led to the higher perox-
idase and laccase activity in comparison with fresh
biochar, which is due to the capacity for enzyme
adsorption and substrate decrease by time (Gibson
et al. 2016).
Biochar properties have been found to be the most
important value for enzymes catalysing electron
transfer. In the first place, the presence of an electron
donor (phenolic) and electron-accepting (quinone)
functional group on the biochar surface is noted. This
allows biochar to be considered as an electron
medium. It has been shown that biochar is able to
accept the electrons formed in the case of acetate
oxidation by the bacterium Geobacter sulfurreducens
(Yu et al. 2016), which participates in restoring the
dechlorination of pentachlorophenol (Yu et al. 2016).
At the same time, the biochar donor-accepting prop-
erties depend on the initial substrate and the pyrolysis
temperature to a high extent. Biochar electron donor
properties increase from 650 to 800 °C (due to
conjugated systems of p-electrons in aromatic struc-
tures) (Zhang et al. 2018c).
Lately, more data have appeared about the role of
electron extracellular transfer among various types of
soil bacteria (Holmes et al. 2017), including those that
involve biochar (Lovley 2017). It has been discovered
that biochar application promotes electron transfer
among microorganisms. Electrons may also be trans-
ferred between bacterial cells and soil minerals,
containing metals of variable valence, especially iron
and manganese (Shi et al. 2016); it has also been
shown that biochar is involved in such electron
transfers (Kappler et al. 2014). Biochar capacity
taking place in electron transfer may form the basis
of the dehydrogenase activity increase, observed
because of its participation in the electron transfer
onto the substrate (in vitro conditions-on triphenylte-
trazolium chloride TTC). One of the possible mech-
anisms of catalase and peroxidase activity increase
Environ Geochem Health
following biochar application is the microbial com-
munity reaction to oxidative stress. It has been shown
that various inductors of oxidative stress lead to a
laccase activity increase (Crowe and Olsson 2001).
Oxidative stress and the induction of enzymes may
be associated with the presence of persistent free
radicals (PFRs) on the biochar surface. Such radicals
are always present on biochar surface, and their
formation is closely linked to the pyrolysis conditions
observed in the recent survey (Qin et al. 2018).
Obviously, for biochar production with a low pyrol-
ysis temperature, a smaller PFR number is formed,
which provides the induction of antioxidant protection
enzymes. At a high pyrolysis temperature, PFR
content on biochar, as a rule, increases (Liao et al.
2014), which may lead to toxic effects for microbial
communities and decrease the enzymatic activity.
Besides, PFR content on biochar depends not only on
temperature but also on the pyrolysis time (Fang et al.
2015), which is not always taken into account in
experiments using biochar. It has been stated that PFR
on biochar surface may take part in electron transport
in the reduction of organic compounds (Gong et al.
2014). This allows us to suggest their influence on test
results for determining dehydrogenase activity. It is
recommended to pay special attention to the choice of
conditions (pyrolysis time and temperature), and even
more to determine PFR content on the surface of
biochar taking part in the experiments at this connec-
tion when planning experiments to determine oxi-
doreductase activity.
Biochar interaction with bacteria in heavy metal
detoxification
Biochar is widely used in the remediation of soils
polluted by heavy metals (HM) due to its sorption
properties. It was studied the biochar effect on the HM
accumulation in the soils in the field conditions as well
as in laboratory conditions. All data received from
above-mentioned studies were published in the review
articles and meta-analyses (O’Connor et al. 2018;
Lwin et al. 2018; Chen et al. 2018a, b). It was
established that Cu and As were mobilized, but Cd and
Zn were immobilized in soils with hardwood-deri-
ved biochar amendment in comparison with un-
amended soil (Beesley et al. 2010). The biochar
application to the soil produced from the rice straw
as well as bamboo wood led to a significant reduction
in the Cd, Co, Pb and Zn mobile forms (Lu et al. 2017).
At the same time, the biochar using led to the
transformation of HM mobile forms into the less
mobile carbonate forms, which produced a long-term
effect from its application into the soils polluted by
lead and zinc near mining enterprises (Meng et al.
2018). In the soils polluted by lead, the pH increase
associated with biochar application led to HM
bioavailability decrease and a positive impact on
microbiological processes (Igalavithana et al. 2017).
Similar data on HM bioavailability decrease associ-
ated with a pH increase at biochar application were
obtained for copper, zinc and cadmium (Lu et al.
2017). Biochar application together with compost in
the vineyard soils reduced Cu2? concentration, which
is the copper fraction, in the soil solution causing most
of the ecotoxicological effects. This effect was
especially expressed in acid soils (Soja et al. 2018).
From the general data, it can be concluded that the
biochar is the most effective HM sorbent in the soil.
However, its potential for environmental pollutant
removal depends on many factors such as raw
materials for biochar production, pyrolysis tempera-
ture and velocity, soil properties, and the application
dose. There are fewer data about biochar interactions
with microorganisms in the HM sorption process and
about the microorganisms’ impact on this process.
The majority of researches are associated with
biochar application’s impact on microorganisms in the
soils polluted by HM and focused on the reduction of
HM toxic effects in natural communities. Thus, it has
been shown that biochar from vegetable waste is
capable of sorbing lead from contaminated soil, which
leads to an increase in microbial biomass in the soil
and an increase in enzymatic activity (Igalavithana
et al. 2017). Application into the soil polluted by Cd
and Pb of 5% of Macadamia nut shell biochar reduced
metal toxicity for the soil microbial community that
resulted in biomass growth and increased the effec-
tiveness of carbon immobilization by microorganisms
and, as a consequence, the reduction of the carbon
dioxide emission into the atmosphere (Xu et al. 2018).
There are data about a significant impact of the
microorganisms on the biochar sorption properties that
is more completely expressed in the case of co-
sorbents’ use. Particularly, in the latest study (Wang
et al. 2018a, b, c) it was shown that the biochar from
the corn stems and pig’s manure, inoculated by metal
123

resistant Bacillus subtilis strain biomass, demon-
strated a higher sorption capacity to Pb and Hg
solution mixture in comparison with uninoculated
biochar. At the same time, the use of the pure lead salt
solution led to the increase in sorption capacity and in
the case of mercury salt solution led to its reduction.
This is connected to the change in HM adsorption on
the biochar surface caused by inoculation with the
bacteria biomass (Wang et al. 2018a, b, c). The use of
bacteria for the joint biochar application into the soils
to reduce the HM toxic action is rather effective. At the
same time, in many works, a considerable positive
effect has been observed in comparison with their use
for the soil remediation separately. In the model
experiment with the soil pollution by Cr(VI) salts, the
best results in phytotoxicity decrease were obtained
only in the case of a joint 5% biochar and biomass
consortium application from two types of bacteria:
Bacillus cereus and Pseudomonas japonica. It was
shown that roots, stems, fresh and dry biomass length
increased up to 135, 168, 370 and 390%, respectively,
in comparison with the polluted soil (Arshad et al.
2017). It can be supposed that the synergetic effect of
biochar and bacteria on the Cr toxicity decreases
caused by the reduction of Cr(VI) to Cr(III). The
stimulation of the plant growth by using the bacterium
Neorhizobium hautlense together with biochar in
agricultural soils polluted with Pb and Cd led to a
considerable decrease in the content of these pollu-
tants in the edible parts of plants (Chinese cabbage and
radish). At the same time, the HM mobile content in
soil rhizosphere was also completely decreased (Wang
et al. 2016a, b). It has been shown that when
introduced into the soil together with the biochar, the
strain of bacteria Pseudomonas sp. with high ability to
reduce Hg(II) increased the efficiency of this process
and accelerated the removal of Hg(II) from the soil
(Chen et al. 2018b). It should be noted that the HM
detoxification mechanisms for the joint use of biochar
and microorganisms remain unstudied. Nevertheless,
the data available in the literature allow us to suggest
that the basis of some of them may be a direct biochar
participation in HM transfer into the less toxic forms.
At the same time, the mechanisms of biochar use as an
electron shuttle described in the literature for organic
pollutant reduction al. are not excluded (Yu et al.
2015). Another possible mechanism is the change in
superficial properties of biochar itself rather than the
123
Environ Geochem Health
microorganism cell adsorption on it (Wang et al.
2018a, b, c). Finally, in the process of biochar and
bacteria interaction, HM ion transfer probably takes
place first from the soil solution into the cells and then
their active pumping out of bacteria cells with their
following sorption on biochar (Fig. 1). The mecha-
nism of active extraction of toxic metal ions from the
cytoplasm is one of the main mechanisms of bacterial
resistance to HM. As shown for Ralstonia sp., metal
precipitation may occur in the form of carbonates due
to CO2 emission by the cells during respiration (Nies
2000). This process affects the composition of the soil
solution, dissolution and migration processes, metal
precipitation and ion exchange processes (Pinskii et al.
2014; Nevidomskaya et al. 2016).
A—direct biochar electron transfer with HM
reduction to less toxic forms, B—forming new
adsorption sites due to colonization of biochar by
bacteria, C—active transport of HM ions from the
bacterial cells and simultaneous carbonates formation
due to cell respiration.
The above-mentioned mechanisms could partially
explain the synergic effects from biochar and bacteria
joint application described in the literature, but
supplementary studies are necessary for their exper-
imental validation.
Biochar interaction with bacteria
in the detoxification of stable organic pollutants
Besides HM sorption and their bioavailability, there
are many studies on biochar participation in stable or-
ganic pollutant degradation. Among them, a special
place is occupied by PAHs that are widely spread
throughout soils of various types. The problem of
PAHs formed at the production of biochar itself and
their impact on microbial communities have been
considered above (Koltowski and Oleszczuk 2015;
Keiluweit et al. 2010).
Two main approaches of biochar utilization for
organic pollutant destruction may be given as natural
microorganism community activation, and the second
one consisted in soil self-clearing process stimulation
by using the biochar application. In the soils polluted
by PAHs and HM, biochar application promotes PAHs
destruction due to HM adsorption (Wawra et al. 2018).
Environ Geochem Health

The studies of the soils polluted by petroleum considerable increase in phenanthrene, fluoranthene
products showed that biochar from sawdust and wheat and pyrene degradation in the soil historically polluted
straw application led to a considerable decrease in by PAHs in comparison with biochar application alone
PAH content. Biochar obtained at a high pyrolysis or bacterial biomass alone (Xiong et al. 2017). A
temperature (500 °C) was more effective than that supposed mechanism is PAHs adsorption on biochar
obtained at 300 °C; also, the initial raw material did with the following biodegradation of immobilized
not decrease PAHs content. Overall, the biochars bacteria. The use of microbial consortiums kept in
produced at higher temperatures exhibited the higher cases from alginate adsorbed on biochar allowed to
sorption efficiency for organic contaminant remedia- effectively reduce soil toxicity at a combined pollution
tion in soil. This is probably due to the high surface with Cr(VI) salt and pyrene (Wang et al. 2018a, b, c).
area and microporosity of biochars (Uchimiya et al. Mechanisms of biochar interaction with microor-
2010). A positive correlation between micropore ganisms at persistent organic pollutant degradation
volume and surface area suggests that the pore size remain unknown, but, obviously, besides sorption they
distribution is a key factor responsible for the increase involve the chemical interaction of pollutants with
in surface area in biochar (Downie et al. 2009; Ahmad biochar that may increase their availability for bacte-
et al. 2014). At the same time, considerable changes rial destructors. Such an interaction is likely associated
were observed in the soil in the microbial community with free radical mechanisms (Fang et al. 2014).
composition with an increased share of PAHs- In particular, it has been shown that PFR on biochar
degrading taxons (Kong et al. 2018). In the compar- surface may interact with hydrogen peroxide, causing
ison of biochar from bamboo (Bambusa vulgaris) hydroxyl radical formation, which degrades 2-chloro-
obtained at 700 °C and biochar from corn stems (Zea biphenyl (Fang et al. 2014). For rice shell biochar
mays) obtained at 300 °C, the first showed reduced
PAHs bioavailability due to adsorption, while the
second stimulated PAHs degradation activation using
microbial communities of carrot rhizosphere (Daucus
carota). At the same time, the proportion of bacteria
belonging to Arthrobacter and Flavobacterium genera
increased and the number of genes associated with
PAHs degradation was augmented (Ni et al. 2017).
Similar results were obtained by the same group on
rice field soils (Ni et al. 2018), which allows a leading
role of biochar type (raw materials and pyrolysis
conditions in the following interaction with microor-
ganisms destructors) to be proposed. Probably, the
biochar with low pyrolysis temperature will interact
with the PAHs very closely and make them inacces-
sible, not only for the plants but for the microorgan-
isms as well. It has been noted that biochars obtained
at high pyrolysis temperatures are the most effective in
absorbing of non-polar pollutants due to a high
structure aromaticity (Ahmad et al. 2014).
The second approach is to use biochar as a carrier of
microbial destructor strains. Pseudomonas putida
immobilized on biochar differing in initial raw
materials and pyrolysis temperature led to a significant
increase in PAHs degradation. The effect for the 4–5-
ringed PAHs was stronger than for 3- and 6-ringed
ones (Chen et al. 2012). Mycobacterium gilvum Fig. 1 Possible mechanisms of biochar and bacteria joint
immobilized on rice straw biochar led to a action in heavy metals’ detoxification

123

Fig. 2 The supposed mechanisms of biochar interaction with microorganisms in the soil
obtained at 550 °C, the mechanism of oxygen active
form generation has been shown leading to chloroben-
zene degradation (Zhang et al. 2018a, b, c). In
addition, the data about biochar PFR participation in
the process of p-nitrophenol degradation in the
experiments without microorganism participation
were obtained (Yang et al. 2015). However, it is
highly likely that these mechanisms could take place
in natural conditions as well. There are also data about
the accumulation of hydrogen peroxide in the media
during incubation of the PAH-degrading strain Achro-
mobacter xylosoxidans with various PAHs (Sazykin
et al. 2018). Further interaction of hydrogen peroxide
with biochar PFR may lead to hydroxyl radical
formation, causing the destruction of PAH aromatic
structures that increases their availability for bacterial
transformation.
The interaction of biochar with microorganisms is a
complex system, which is influenced by both the
123
Environ Geochem Health
features of the carbon sorbent itself and the properties
of the soil in which it is introduced. Under conditions
of soil contamination with heavy metals and polyaro-
matic hydrocarbons, this system becomes even more
complex. At the same time, all elements of the system
interact with each other through various physico-
chemical and biological processes. Biochar interaction
with microorganisms and soil is shown in Fig. 2.
Conclusions
Biochar utilization for the improvement of agricultural
soil properties and as an ecologically safe sorbent for
the polluted soil immobilization has significant poten-
tial. The effectiveness of its use to a considerable
extent depends on the pyrolysis conditions, the biochar
precursors and soil properties. The addition of biochar
can effect the soil properties to a large extent. There is
Environ Geochem Health
no unambiguous answer to the question about the
impact of biochar on the soil structure. Some studies
demonstrated the soil aggregation, while others
reported no effect at all. Much remains to be learned
about the changes in sorption properties of the biochar
during its colonization by microorganisms, as well as
about the mechanisms of the new adsorption sites
formation on the surface of carbon granules.
Concerning biochar interactions with microorgan-
isms in the remediation of HM- and PAHs contami-
nated soil, microorganism participation in this process
remains under-explored. With regard to the use of
biochar for the remediation of soils contaminated with
organic pollutants, both the stimulation of microor-
ganisms already presents in the soil and the develop-
ment of a new generation of co-sorbents using biochar
as a carrier of highly active degrader strains will be
promoted in the future.
Changes of pH caused by biochar application and
biochar potential toxicity due to volatile pyrolysis
product emissions play the leading role in the impact
on soil microbial communities’ functioning and com-
position. The difference in soil properties and the
complexity of biochar impact on the soil can lead to
contradictory data and the difficulty of comparing the
results of experiments performed. Particularly, many
gaps exist in realizing the mechanism of biochar
interaction with the soil microbial community.
For a better understanding of biochar impact
mechanisms on the soil and microorganisms, it is
necessary to develop a single model of biochar testing
including the list of parameters that must be studied.
Before the start of widespread biochar use in agricul-
tural practice, it is necessary to develop the interna-
tional standards on potentially toxic pyrolysis products
content as well as the methods of removing potentially
negative effects by means of pre-treating of biochar.
The further study of biochar interactions with
microorganisms and their combined deployment in
soil will allow the application of a highly effective and
ecologically safe instrument for soil remediation, if
certified biochar of high quality is used.
Acknowledgements This research was supported by projects
of Ministry of Education and Science of Russia, No.
6.6222.2017/8.9 BP, Grant of the President of Russian
Federation, No. MK-2973.2019.4, RFBR No. 19-29-05265 mk.
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