Journal of Cleaner Production 262 (2020) 121372

Contents lists available at ScienceDirect

Journal of Cleaner Production

journal homepage: www.elsevier.com/locate/jclepro

Review

Microbiologically Induced Calcite Precipitation biocementation, green

alternative for roads e is this the breakthrough? A critical review
Carla Ribeiro Machado e Portugal a, *, Carolyn Fonyo a, Carlos Cardoso Machado b,
Richard Meganck c, Todd Jarvis d
a
Environmental Sciences Graduate Program, Oregon State University, Graduate School, Heckart Lodge, 2900 SW Jefferson Way, Corvallis, OR, USA
b
Federal University of Viçosa, College of Forest Engineering, Avenida Peter Henry Rolfs, S/N, Viçosa, MG, 36570-900, Brazil
c
Institute for Water and Watersheds, Part of the UNESCO-ICIWaRM Category II Center, Oregon State University, USA
d
Institute for Water and Watersheds, Strand Agriculture Hall 230, 170 SW Waldo Place. Corvallis, OR, 97331, USA

a r t i c l e i n f o a b s t r a c t

Article history: The past fifteen years have been a rich developing field for Microbiologically Induced Calcite Precipita-
Received 14 October 2019 tion (MICP) as an alternative to decrease the hydraulic conductivity of sandy soils or fractured rocks
Received in revised form through biocementation formation. While significant development happened, the field still lacks a viable
20 March 2020
answer for the use of MICP for organic soil stabilization. Sandy soils or fractured rocks have a completely
Accepted 25 March 2020
Available online 31 March 2020
different structure and mechanical behavior than organic soils, which can impact the behavior of bio-
logical components and MICP microorganisms. This factor per se changes all the research perspective
Handling editor. Prof. Jiri Jaromir Kleme
s and demand adaptations different than the ones used for sandy soil or fractured rocks. In the search for
answers, this article compiles an extensive and systematic literature review with papers and books from
Keywords: platforms like Academic Search Premier, Web of Science, Google Scholar, 1Search and Gale Virtual
Microbiologically induced calcite -Reference Library, having the selection based on defined parameters and criteria. Focusing on how MICP
precipitation (MICP) is a potential solution for unsealed road stabilization, this article discussed the main gaps and constraints
Ureolytic bacteria that could explain why biocementation still needs extensive research under different perspectives and
Unsealed roads
scenarios. The results suggested that the majority of investigations are at a similar stage of limitation:
Biocement
how to guarantee an evenly spread of biocementation into organic soil at large extensions. The dis-
Organic soil
Soil stabilization cussion here provided some insights and pieces that can enrich future researches, support the expansion
of the development, and improve the understanding of the biocementation process into organic soils and
its use for unsealed road stabilization.
© 2020 Elsevier Ltd. All rights reserved.

Contents

1. Introduction - from biomineralization to biocementation to improve the soil for roads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

2. Research methodology . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.1. Literature research . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
2.2. Screening and papers selection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Results & discussions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3.1. Soil bacteria as the biocementation agent . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3.2. Biocementation & MICP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.3. Gaps, constraints, and the uses for MICP - a brief microbiology perspective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
3.3.1. Understanding MICP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.3.2. Urease (UE): is this a limiting factor for the MICP? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

* Corresponding author.
E-mail addresses: [email protected], [email protected]
(C.R.M. Portugal), [email protected] (C. Fonyo), [email protected]
(C.C. Machado), [email protected] (R. Meganck), Todd.jarvis@oregonstate.
edu (T. Jarvis).

https://doi.org/10.1016/j.jclepro.2020.121372
0959-6526/© 2020 Elsevier Ltd. All rights reserved.
2 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

3.3.3. - Carbonic anhydrase (CA): is this another limiting factor? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

3.3.4. The nucleation sites and their impacts on precipitation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
3.4. Nitrogen cycle and ammonium concentration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
3.4.1. Precipitation of calcium carbonate crystals and their relevance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4.2. The potential biohazard of exogenous bacteria for soil stabilization: biological or economic concern? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.4.3. MICP as a potential source of carbon sequestration: adding value on unpaved roads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.5. Main soil parameters for a successful biocementation process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.5.1. Permeability (Hydraulic Conductivity) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3.5.2. Soil stratification & permeability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.5.3. Porosity (void fraction) and pore size distribution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.5.4. Shear strength . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.5.5. Soil microstructure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
3.6. MICP on unsealed roads . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.6.1. Granulometric stabilized soil - the foundation for unsealed road construction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.6.2. Soils stiffness and stabilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.6.3. Erosion on unsealed roads: chemical stabilization methods and MICP . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
3.6.4. Natural calcite precipitation in soils - a prediction model . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
4. Conclusions & considerations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Funding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Declaration of competing interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

1. Introduction - from biomineralization to biocementation stabilization processes, including unsealed road surfaces and dams.
to improve the soil for roads Biostabilization consists of improving the geotechnical properties
of soils by Microbially-Induced Calcite Precipitation (MICP), the
Biomineralization is the precipitation of crystals (organized precipitation of calcium carbonate from ureolytic bacteria activity
shape mineral) in the cellular or extracellular matrix of a living (Whiffin, 2004; Van Paassen, 2009; DeJong et al., 2013). The cal-
organism (Lowenstam and Weiner, 1989). It is a widely known cium carbonate precipitation (or biostabilization process) is the
process wherein microbes move different metals to form minerals result of a microbial metabolic process such as photosynthesis, and
(Dhami et al., 2013, 2016; Phillips et al., 2013; Zhu and Dittrich, hydrolysis of urea and reduction of sulfates, among others
2016). This multistep process is natural for many microorganisms (Valencia, 2009; DeJong et al., 2011; Valencia et al., 2014). This
and animals that precipitated various minerals, like iron oxides in article focuses on review the MICP process, where the precipitate of
magneto bacteria, magnesium silicates in crustaceans, and calcium CaCO3 can biocement soil particles and reduce its hydraulic con-
carbonates and calcium phosphates in invertebrate shells and ductivity (Whiffin, 2004; Umar et al., 2016; Van Paassen, 2011).
vertebrate skeletons (Boskey, 1998). The resulting biominerals de- Calcium Carbonate is an attractive element for the biostabilization
posits in elaborated shapes and hierarchical structures based on the process since the calcite formation is a natural process. Several
organic-inorganic interface. The rate of crystal formation controls authors such as Stepkowska et al. (2003), Bessler and Rodrigues
the microenvironment and the mineralization. One of the most (2008), Yang et al. (2009), Zhang et al. (2010), Wei et al. (2015),
studied biomineralization processes results in calcium carbonate €
Ozen and Simsek (2015); Piekarska et al. (2017); Donnelly et al.
(CaCO3), as Calcite, one of the most stable forms (Ferna ndez et al., (2017); Jaji et al. (2017) summarize that Calcium Carbonate Min-
2018). erals (CCM) are present in nature under one of its three crystalline
New awareness in the role of microbial processes in bio- forms - vaterite, aragonite or calcite. When the CCM’s are under the
mineralized geological formations has created the interest of re- trigonal or rhombohedral crystalline structure, they are called
searchers worldwide (Banks et al., 2010; Ronholm et al., 2014; Calcite. Having Calcite as the most stable form of calcium carbon-
Rusznya k et al., 2012). There has been a growing interest in the ates, thus being the target of majority MICP’s research.
exploration of novel microbes and routes which have the similar The most studied MICP microorganisms are urease-positive
ability to precipitate desired compounds as calcium carbonates to bacteria. Since MICP bacteria are common in the soil environ-
fix or mobilize metals, sequester atmospheric CO2 for applications ment, many studies indicate the probability of a successful in situ
in different engineering areas (Dhami et al., 2013, 2014). biostabilization treatment of sand or sandy soil with ureolytic
Meldrum (2003) classifies biomineralization as “biologically bacteria (Whiffin, 2004; Umar et al., 2016; Whiffin et al., 2007; Van
induced” mineralization or “organic matrix mediated mineraliza- Paassen, 2011; Mujah et al., 2017; Bibi et al., 2018). The majority of
tion”. “Biologically induced” mineralization is the interaction be- the research, including the cited researchers above, focused on
tween an organism and its environment under low biological Bacillus spp and Sporosarcina spp, which can be exotic bacteria
control of the mineralization process, resulting in precipitation of a depending on the location. The bacterium death reduces the risk of
biomineral. “Organic matrix mediated mineralization” has the live gene escape due to physical isolation and lack of nutrients. How-
organism directly controlling the biomineralization process. This ever, Hokkanen and Lynch (2003) reported that predicting the
article focuses on the “biologically induced” mineralization branch, behavior of exotic bacteria introduced into the soil must be
limiting on Microbiological Induced Calcite (CaCO3) Precipitation analyzed on a case-by-case basis, as it involves specific kinetics
(MICP) process. principles for each species. The same authors stated that the gen-
Several biomineralization research projects and methodologies eral trend is the decline in the population density of bacteria
are under development and evaluation, including biostabilization introduced in a short time (15e25 days for common ones). They
or biocementation, which is a robust green alternative for soil conclude by stating that competition against bacteria already in the
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 3

soil and predation by other microorganisms represent the most research. The first stage was an extensive search of five databases
significant potential impacts on the new bacterial population. approved by Oregon State University: Academic Search Premier,
In summary, the authors stated that the establishment of a Web of Science, Google Scholar, 1Search, and Gale Virtual Reference
bacterial population depends on the soil-bacterial interaction, with Library. The relevant literature was selected based on specific
significant weight in the bacterial sepa, determining a highly spe- keywords including MICP, Microbiologically Induced Calcite Precipi-
cific interaction, classified as soil-dependent interaction. Even tation, Microbially-induced calcite precipitation, calcite, Calcium
though the use of native bacteria can potentially reduce the cost of carbonate, biostabilization, biocementation, biocalcification, biogrout,
using imported and patented products, the authors could not find urea, urease, ureolytic bacteria, native bacteria, nucleation site, shear
any analyses about the risk-evaluation of the potential biohazard strength, permeability, porosity, soil stabilization for roads, unsealed
contamination from exogenous bacteria proposed for soil stabili- roads, and MICP, unsealed roads and biocementation, biological sta-
zation. This lack of information indicates the first gap of the MICP bilization for unsealed roads. The selection and combination of this
field, evaluated in this paper. set of keywords were defined to allow the identification of mean-
This manuscript presents a detailed review of significant pub- ingful literature and avoid biased research. The combination of
lications related to MICP as a soil stabilization technique. It dis- keywords was determined to cover the topics discussed here. The
cusses the main critical gaps and constraints for MICP as a green second stage of the study was the cross-reference search in the
alternative for soil stabilization: how to ensure homogeneous and articles with the highest citation index selected in phase 1, to
widespread biocementation without compromising the shear complement the search with more recent papers. Also, we
strength or durability of the soil surface and its extension for road retrieved the non-detected relevant papers from the bibliographic
stabilization. Many publications demonstrate the MICP process on references from the first stage of the research. The initial selection
the sand and sandy soils, but this review focused on organic soils, process identified 2232 articles as potentially appropriate to the
which is a topic in early development at MICP technology. These topics selected for discussion in this article.
factors, allied with the urgent need of a green solution for unsealed
road erosion, guided the research questions of this article: a) Which 2.2. Screening and papers selection
is the current state of the art of MICP on organic soils?; b) What are
the possible gaps and restrictions in the behavior of ureolytic The papers search process based on several citations and
bacteria, agents of MICP, for organic soils?; and c) What are the gaps renowned researchers in the areas of MICP, microbiology related to
and constraints for the use of indigenous bacteria in the MICP the behavior of ureolytic bacteria, and unsealed roads. Table 1
process on unsealed roads? This review brings more information shows the criteria for the selection of articles used in this review.
about MICP and its intrinsic and unique interaction with organic Please note that had no limitation on publication date as we seek to
soils, promoting a focused debate about the gaps and constraints of consolidate understanding of the entire MICP process to identify
the homogeneous application and resistant biocementation. The the insights that can support new research and ideas in the search
MICP process can not compromise the shear strength or durability for solutions to MICP restrictions and gaps in road stabilization
of the unsealed road (granulometric stabilized soil, a composite of unsealed.
gravel, sand, silt, and clay). The first selection evaluated only the titles and abstracts,
generating a total of 1124 selected articles. Although this is a large
2. Research methodology volume, the next stage evaluated the discussion and conclusion of
the papers, reducing it to a final selection of 158 papers.
This review article compiled an extensive range of papers that
collect details and information about the MICP process, soil stabi- 3. Results & discussions
lization, unsealed roads, and above all, why the ureolytic bacteria
can be a potential solution for biocementation of unsealed roads. 3.1. Soil bacteria as the biocementation agent
This systematic literature review, conducted under a holistic
approach, sought to condense information about the possible gaps Soil microbe communities form one of the most abundant mi-
and constraints in the development of the protocol for the use of crobial ecosystems on Earth, led by bacteria species (Mitchell and
biocementation via ureolytic bacteria. The protocol will target Santamarina, 2005; Wang et al., 2020; Coleman-Derr et al., 2016;
construction and management of unsealed roads in tropical areas, Wu et al., 2006; Umar et al., 2016). Some of these bacteria species
indirect benefit the conservation of water resources, which at least biomass is present in large numbers than others since the biotic
would no longer receive sediment from the erosion process of and abiotic factors that affect the fitness of these microorganisms
unsealed roads. varies across the depth of the lithosphere (Hokkanen and Lynch,
2003; Umar et al., 2016).
2.1. Literature research Madigan (2012) reported that bacteria could survive in envi-
ronments with the most varied rates of acidity, salinity, tempera-
Many authors published reviews about MICP, having the most ture, and atmospheric pressure. Most bacterial species survive in
cited ones from Whiffin (2004), Whiffin et al. (2007), Van Paassen places with pH values between 5 and 7, which is typical of
(2009, 2011), Anbu et al. (2016), Seifan and Berenjian (2019). groundwater and soils close to the surface; and the pH decreases
However, the literature lacks its applicability at large scale, or with the increase of the concentration and valence of ions found in
organic soils left gaps that need answers. Some few articles the fluids present in the soil (Madigan, 2012; Chapelle, 2000).
describe the MICP procedure for organic soils but do not have Ureolytic bacteria produce the urease enzyme, which indicates
compiled information, nor clear described protocols. Whiffin its potential use in the biomedical soil improvement technique -
(2004), Whiffin et al. (2007), and Soon et al. (2013) helped shape MICP (Kucharski et al., 2006). The genera of bacteria most
this review. The first authors were one of the pioneers on research commonly used in MICP techniques are Bacillus, Sporosarcina,
MICP as biocement for soil improvement, and the second authors Spoloactobacillus, Clostridium, and Desulfotomaculum (Kucharski
brought some of the first insights about how to improve the et al., 2012) and, more recently, the genus Serratia, a bacterium
technique with organic soil. native to Brazilian soil with research under development (Enriquez,
This present review adopted a two-stage search for literature 2017).
4 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

Table 1
Inclusion and Exclusion Criteria per parameter for proper paper selection for this study.

Parameter Criteria

Inclusion Exclusion

Publication type Journal paper, thesis, dissertations, book chapters, and conference paper Editorial and comments
Language English or Portuguese Other languages
Accessibility to the full Available Not available
text
Topics 1. MICP process for diverse materials, except those with the biological and medical 1. MICP process for biological/medical purposes
purpose 2. MICP for other purposes
2. MICP as a source for biocement 3. Biocement for construction and art repair in
3. Biocement for soil stabilization details
4. Soil stabilization for Unsealed road construction 4. Soil stabilization for sealed road construction

Morales et al. (2015) tested a biocementation scenario with a Mitchell et al., 2013); pollutant compound immobilizers (Mitchell
low amount of biocementing solution, simulating the levels natu- and Ferris, 2005; Fujita et al., 2008), concrete self-healing
rally produced by a bacterium (undeclared) from the Bacillaceae (Jonkers et al., 2010) and potential CO2 sequestration (Mitchell
family in its original condition of development in silty-clayey- et al., 2010; Cunningham et al., 2014; Phillips et al., 2016), and
sandy soil. The objective was to limit the production of ammonia more recently, the innovative proposal for stabilizing unsealed
and its oxidized forms to a low level to minimize the potential roads, with biocementation as a potential substitute for chemical
environmental impact and the possible dissolution of precipitated stabilization with cement or lime (Enriquez, 2017) and improve-
calcium carbonate due to acidification of the medium by oxidizing ment of soil engineering properties (Minto et al., 2017; Osinubi
ammonia. The results showed a low level of precipitation, less et al., 2020). It has been showed that biocementation could suc-
resistance to compression, and the most moderate shear strength cessfully enhance the strength and stiffness of pure sand in a
when compared with other tests for soil stabilization. The authors relatively large area of extension (Van Paassen et al., 2010, Ivanov
concluded that induced biocementaça ~o the soil with a low inci- and Chu, 2008; DeJong et al., 2010, 2013; Dhami et al., 2013; Gao
dence of only precipitation can only be used for purposes of soil et al., 2019, He et al., 2020). However, it was not possible to repli-
filling that do not require high shear strength. cate this result on silty sandy, silt, or clay soil (He et al., 2020).

3.2. Biocementation & MICP 3.3. Gaps, constraints, and the uses for MICP - a brief microbiology
perspective
Microbial induced carbonate precipitation (MICP) is a natural
process, controlled by different mechanisms (Banks et al., 2010; The MICP or biocementation technology is on the verge of a
Cacchio et al., 2003; Wright and Oren, 2005). One of these mech- breakthrough, which is a short period could lead to the expansion
anisms is the production of calcite by ureolytic bacteria in porous of biocement application in a variety of environmental scenarios.
soil, when in the presence of urea and calcium ions (Stocks-Fischer Many researchers worldwide like Santamarina, Chou, DeJong,
et al., 1999; Frankel and Bazylinski, 2003; Whiffin, 2004; Mitchell Montoya, and van Paassen in the USA; Whiffin, Cheng, Premkumar,
and Santamarina, 2005; Ivanov and Chu, 2008; DeJong et al., Al-Thawadi and Ismail in Australia; Al Qabany and Soga in the UK;
2010; De Muynck et al., 2010; Ivanov, 2010). The complete under- Valencia e Enriquez in Brazil, (just to cite some) are devoted to solve
standing of this mechanism is important to understand the other some significant gaps and constraints of the MICP for soil stabili-
biologically induced natural CaCO3 precipitation mechanisms zation. Among the gaps, we highlight: (i) How to obtain and sustain
(Banks et al., 2010; Cacchio et al., 2003; Fukue et al., 2003; Wright a homogenous biocementation layer through the treated area; (ii)
and Oren, 2005) and how to transfer this knowledge to How to get a homogenous biocement production despite the media
commercial-scale production of the MICP in its various uses. (sand, soil, or any soil mix), (iii) How to maintain homogeneous
The MICP process depends on six main factors: (1) concentra- distribution of the nucleation sites to allow precipitation volume
tion of calcium, (2) level of dissolved inorganic carbon (DIC), (3) pH that can lead to a strong biocementation? Until questions remain
of the medium, (4) availability of nucleation sites, (5) urease ac- open, it is not possible any advancement in economic analysis and
tivity; and (6) carbonic anhydrase activity (Hammes and Verstraete, environmental assessments due to few studies at field scale that
2002; Hammes et al., 2003; Achal et al., 2015). MICP involves a can support the conclusions.
series of complex biochemical reactions that can be affected mainly All the MICP papers consulted have similar constraints that
by ambient temperature, pH, water content, urea concentration, support our central question: Does the current level of the MICP
species and concentration of bacteria, pore sizes, and soil void rates homogeneity for soil stabilization, independent of the bacteria
(McConnaughey and Whelan, 1997; Stocks-Fischer et al., 1999; selected, justify or provide robust parameters and protocols to
Whiffin, 2004; Whiffin et al., 2007; Van Paassen et al., 2010; Achal sustain the MICP at a commercial scale? Tests on MICP for soil
et al., 2009a; Harkes et al., 2010; Achal and Pan, 2011; Sharma and stabilization at laboratory scale is still highly used, which can be
Ramkrishnan, 2016; Castro et al., 2016). supported by all these factors cited above and rationale of authors
The MICP process is highly dependent on active ureolytic bac- like Al-Thawadi (2008), De Muynck et al. (2010), DeJong et al.
teria, sources of calcium chloride and carbon, source of urea, level of (2013), Dhami et al. (2016); Umar et al. (2016) among others. This
urease that increases the pH, promoting rapid precipitation of scale can reduce the costs, potential environmental impact, and
calcium carbonate (Ferris et al., 2004). The main lines of research simplify research efforts promoting a substantial advancement to
for the MICP focus on improving the strength and stiffness of the field.
porous media while maintaining permeability (Whiffin et al., 2007; Even though the biocementation homogeneity is the most sig-
Van Paassen, 2009; DeJong et al., 2010); reduction of permeability nificant gap, we have to consider two main constraints for any MICP
in porous media (Tobler et al., 2012; Handley-Sidhu et al., 2013; research. First, potential biohazard from the proposed use of
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 5

exogenous bacteria. Second, ammonium concentration (limiting cereus. However, Enriquez (2017) found that Serratia ureilytica sp
factor) and ammonia production (undesired byproduct) that can has higher precipitation rates of CaCO3 (calcite) when compared to
lead to potential environmental impact, including overall water Sporosarcina pasteurii in sandy soils under the same parameters
quality at the adjacent streams and water bodies. and the same protocol, based on Whiffin (2004). Allied to the fact
that it is a native species, Serratia ureilytica sp is a strong candidate
3.3.1. Understanding MICP for investigations of MICP as a soil stabilizer in Brazil.
In summary, the MICP starts with urease activity, where (stage
1) 1 mol of intracellularly hydrolyzed urea (CO(NH2)2) results in 3.3.3. - Carbonic anhydrase (CA): is this another limiting factor?
1 mol of ammonia (NH3) plus 1 mol of carbonate (NH2COOH), fol- Carbonic anhydrase (CA), like urease, plays a vital role in the
lowed by (stage 2) spontaneous hydrolysis that form 1 additional MICP process, but it still needs more detailed studies on how it
mol of ammonia (NH3) and 1 mol of carbonic acid (H2CO3). In works at MICP. CA is also a metalloenzyme, but during stage 9 uses
aqueous media (stage 3), the carbonic acid and ammonia will reach zinc in the catalytic nodes with carbon dioxide (CO2) and bicar-
equilibrium, forming 1 mol of bicarbonate ions (HCO
3 ), 1 mol of bonate (HCO
3 ), replacing stage 4 in the MICP at the urease pathway.
hydrogen ions (Hþ). Then, (stage 4) each 2 mol of ammonia will In the CA pathway, after the production of bicarbonate, hydrogen
combine with 2 mol of water, resulting on 2 mol of ammonium ions (H þ) promote the precipitation of calcium carbonate in the
(NH


4 ) and 2 mol of hydroxide ions (OH ), (stage 5) increasing the form of calcite (stage 10) plus the output of water and carbon di-
pH and shifting the balance of bicarbonate into carbonate (HCO
3 / oxide (Castro et al., 2016).
CO2

3 ). This balance promotes (stage 6) a high influx of calcium ions Hwang et al. (2013) reported the action of CA in biominerali-
and other protons expulsion, driving the bacteria to (Stage 7) zation and the fact that the morphology of calcium carbonate
release calcium outside the cell to survive. At this stage (8), the (calcite) depends on the constant pressure of CO2 and the addition
presence of dissolved inorganic carbon (DIC) as carbonate ions of polymers that affect the growth and nucleation of the precipi-
outside the cell and the expelled calcium ions triggers the reaction tate. Fig. 2 is a schematic illustration of CaCO3 biomineralization in
that culminates (stage 9) with the Calcium Carbonate precipitation the presence and absence of carbonic anhydrase. The CaCO3
outside the cell. precipitated in the presence of CA can take three forms: ellipsoidal,
polygonal, or rhombohedron. The same process in the absence of
3.3.2. Urease (UE): is this a limiting factor for the MICP? CA only precipitates CaCO3 in the form of a rhombohedron, a less
Urease (EU), a member of the hydrolases group, is a nickel- rigid morphology.
containing metalloenzyme that catalyzes the hydrolysis of urea The activities of urease and carbonic anhydrase are interde-
into ammonia and carbonate, initiating the reaction chain of the pendent and guide the stages of Calcite precipitation. The activity of
calcium carbonate precipitation described above (Castro et al., urease depends on the incorporation of nickel in its nucleation site,
2016). Fig. 1 shows a schematic illustration of the MICP of a ure- which is regulated by the chemical reaction between carbon di-
olytic bacterium, highlighting the reactions that occur on the sur- oxide and bicarbonate catalyzed by CA (Jime nez-Lo  pez et al., 2007;
face of the bacteria in addition to a summary of the internal cell Wong, 2015).
balances that promote calcite precipitation. The carbonic anhydrase
can limit the MICP, as discussed in the next section. The most 3.3.4. The nucleation sites and their impacts on precipitation
studied ureolytic bacteria present in the soil are Sporosarcina pas- Bosak and Newman (2003) demonstrated that the nucleation
teurii (formerly Bacillus pasteurii), Bacillus sphaericus, and Bacillus processes drove the formation of microbial carbonate on our planet

Fig. 1. Schematic illustration of the MICP process by urease activity and ureolytic bacteria (adapted from Castro et al., 2016).
6 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

Fig. 2. Schematic illustration of MICP by carbonic anhydrase activity. (compiled and adapted from Castro et al., 2016 and Müller et al., 2014).

for millennia. However, Bontognali et al. (2008) state that the precipitated calcium carbonate, leading to a potential homoge-
nucleation process and its paleontological significance are sources neous layer of biocement once we have a complete understanding
of much controversy among scientists and require further studies. of the process.
Aloisi et al. (2006), corroborated by Bontognali et al. (2008), Obst We highlight the need for more research on the formation of
et al. (2009) and Achal and Pan (2011), summarized that the nucleation sites and their relationship with ureolysis and precipi-
nucleation sites are where the cell surface of microbes secretes tation of carbonates (calcite), which may contribute to elucidate the
Extracellular Polymeric Substances (EPS) and that they have a gaps around the homogeneity and resistance of biocement aimed at
negative electrical charge and the ability to bind to Caþ 2 ions. The stabilizing unsealed roads. Besides, Warren et al. (2001) showed
authors also stated that EPS is vital for the biomineralization pro- that bacterial surfaces could act as models of mineral nucleation,
cess, as they are the key to the initial stage of carbonate nucleation promoting a reduction in the cost of precipitation activation energy.
processes, enhanced by the progressive development of large They demonstrated that CaCO3 precipitation does not occur in the
granules of calcium carbonate with a granular texture. absence of bacteria, indicating that the amount of energy for acti-
The research by Aloisi et al. (2006) contributed to fill the gap of vation and homogeneous precipitation can be a significant barrier.
more detailed knowledge about the processes and activities of the We understand that this barrier is a potential source of process
nucleation site. This study is relevant, as an acceleration and control. Warren et al. (2001) also indicated that the bacteria in-
intensive nucleation of calcium carbonate on the cell surface can crease the precipitation of calcium carbonate, promoting super-
lead to the confinement of the organism and, eventually, premature saturated conditions.
death. The characteristics of the nucleation site define the
morphology of the calcium carbonate precipitates: Calcite, Vaterite,
3.4. Nitrogen cycle and ammonium concentration
or other metastable polymorphs. Studying Desulfonatronum lacus-
tre, a gram-negative sulfate-reducing and carbonate precipitating
MICP can occur via urea hydrolysis, aerobic oxidation, denitri-
bacterium, Aloisi et al. (2006) described a new pattern of nanoscale
fication, sulfate reduction, and other pathways. However, Van
microbial carbonate nucleation. They found that the calcium car-
Paassen et al. (2010) and Achal and Pan (2011) stated that MICP
bonate precipitation occurs in blood cells in regions close to the
via urea hydrolysis (activated by Urease) is the most controllable
microbial cell wall, and the calcification increases during the
carbonate precipitation pathway, and according to Achal and Pan
release of the blood cells into the aquatic environment around
(2011), it reaches the highest level of CaCO3 precipitation with a
microbes. This model can bring more information and tools to
shorter curing period of biocement.
explain the formation of nanospheres and other steps in the MICP
Ureolytic bacteria can be classified into two groups based on
process. However, Bontognali et al. (2008) described that in sulfate-
their response to the ammonium concentration in the environment
reducing bacteria, the EPS is external, which promotes precipitation
(Gat et al., 2014). In the first group, the high concentration of
and, consequently, agglomeration in the external medium of CaCO3,
ammonium suppresses the activity of urease in bacteria, which is of
which leaves the bacteria mobile and rarely being “buried” by the
low interest for MICP purposes. This group includes Bacillus meg-
mineralization process (low biocalcification). Achal and Pan (2011)
aterium. In the second group, the high concentration of ammonium
corroborate Tsuneda et al. (2003), Achal et al. (2009a) and Achal
does not limit the activity of urease, defining this as the group of
et al. (2009b), stating that EPS is essential in the biofilm bio-
interest for MICP. This group includes Sporosarcina pasteurii
calcification process, cell adhesion and even in the capture of
(formerly Bacillus pasteurii) - the most studied MICP bacterium and
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 7

the biological agent of Biogrout® (Umar et al., 2016). Whiffin (2004) potential source for carbon sequestration (carbon dioxide). This
established that the bacteria in the second group are a wise choice model suggests the MICP promotes carbon capture and storage
for soil improvement because the high concentrations of urea are through the use of soluble CO2 as a carbon source in the carbonate
hydrolyzed in the process, reducing the potential release of urea in precipitation in the biocementation process. The authors summa-
the soil and groundwater. However, the process releases ammo- rized recent research that stated that MICP species with alkalin-
nium (NH4 þ) into the environment, which can be detected very izing metabolism may play an active role in CO2 sequestration and
easily as the natural odor of ammonium at the site of the experi- how to act as a potential long-term carbon sink. Castanier et al.
ment (Al Qabany et al., 2012). None of the papers evaluated (1999) described in detail the variety of reactions mediated by
expressed concern about the release of ammonium into the envi- bacteria through an increase in pH and carbonate alkalinity,
ronment. We believe that is more research about the released resulting in carbonate precipitation. Dupraz et al. (2009a) and
ammonium level before any claim that the MICP is an environ- Dupraz et al. (2009b) referred to this as the “alkalinity engine” that
mentally sustainable alternative. promotes the concentration of HCO3 and CO2 by reducing dissolved
CO2 (CO2 (aq.)). The consumption of CO2 (aq.) Favors the capture of
3.4.1. Precipitation of calcium carbonate crystals and their gaseous CO2 (CO2 (g)) in solution, promoting an increase in pH and
relevance rapid conversion into HCO3 and CO2
3 . These events culminate in
Studied by many authors and summarized by Warren et al. mineral trapping through the precipitation of carbonates. Millo
(2001), calcium carbonate has many polymorphs with different et al. (2012) described the interconnection between the final in-
crystalline structures, with calcite, aragonite, and vaterite as the crease in pH and the decay of the precipitation rate due to the HCO3
most common and stable. The authors stated that every polymorph deprotonation and carbonate precipitation. This interconnection
has different stability due to its micromorphology, resulting in indicates a recovery of the alkalinity mechanism since the con-
different reactivities with the surface of its environment. Calcite is sumption of available calcium promotes a decrease in CaCO3
thermodynamically stabilized and has a hexagonal-rhombohedral precipitation.
crystal structure; aragonite is metastable and has an ortho- Warren et al. (2001) stated that calcium carbonate precipitation
rhombic crystalline structure, usually in the form of a needle; and could interfere with biogeochemical cycles, and therefore, have a
vaterite (precursor to calcite and aragonite) is a metastable hex- potential impact on CO2 concentration at the atmosphere or even
agonal crystalline structure in spherulitic or disc-like form. These with reactive transport of radionuclides and trace metals in
authors concluded that identifying the formation of structures and contaminated aquifers. For these reasons, we understand that
the level of reactivity of precipitated calcium carbonate is vital to acknowledge the MICP process in its deep complexity and mech-
define whether this procedure is a potential bioremediation tech- anistic level can bring more information about the potential carbon
nique for aquifer contamination. Li et al. (2013) showed that sequestration by biocementation of unpaved roads.
indigenous ureolytic bacteria can sequester soluble heavy metals
present in soil and groundwater and still survive during the MICP 3.5. Main soil parameters for a successful biocementation process
process. The authors studied the MICP process via urea hydrolysis
in Sporosarcina pasteurii and Terrabacter. They concluded that both Majority of published articles reviewed for this article stated in
are successful in sequestering Ni, Cu, Pb, Co, Zn, and Cd from the their titles that the research focused on how to improve the “soil”
soil, precipitating them under the resistant carbonate composition strength. However, the results reflect the biocementation applica-
to the acid attack of a level similar to that of acid rain, facilitating tions on sand or sandy soil. The analysis of sand, sandy soil, and
the removal of these heavy metals. granulometric stabilized soil demonstrated that the expected MICP
behaviors were very different depending on the used soil matrix
3.4.2. The potential biohazard of exogenous bacteria for soil (distribution of particle sizes). This analysis indicates that it is
stabilization: biological or economic concern? necessary a considerable diversity of studies and techniques to
Fritzges (2005) stated that bacteria are native to the Earth and successfully obtain a range of resistant biocementation applications
therefore have little chance of causing any environmental risk in for unsealed road stabilization.
the future. Umar et al. (2016) use this assumption as a validation for The technique needs to guarantee reliable and replicable results
the potential use of any bacterium anywhere on Earth. Ecosystems on the evaluation of geotechnical engineering properties like
(micro to macro) are unique and have a delicate balance, where any permeability, porosity, stiffness, shear strength, unconfined
change can change the balance and transform the local environ- compressive strength, and a homogenous microstructure (or soil
ment. However, in the face of biological risk concerns, Umar et al. uniformity), parameters of a reliable soil cementation procedure
(2016) mentions that the MICP bacterium must be selected based (Machado et al., 2009). The next subsections discuss the main dif-
on its environmental safety during and after the treatment process, ferences in the studied factors and how they can potentially affect
avoiding the use of genetically modified organisms (GMOs), or the biocementation process.
pathogens or the inclusion of any exchangeable toxic element that
may affect local microbial pathogenicity (indigenous bacteria). 3.5.1. Permeability (Hydraulic Conductivity)
Have to consider this statement in cases like the Biogrout® or other Caputo (1999) defines soil permeability as the capacity of water
exotic bacteria species that lack data behavior in the environment percolation through its particles under different rates, based on its
of each study. In order to minimize the potential environmental composition. The permeability coefficient (k), derived from Darcy’s
impact of exotic species and reduce costs, we suggest the devel- law, correlates discharge and viscosity, where discharge is directly
opment of studies with native (indigenous) bacteria. proportional to soil (media) hydraulic gradient. Permeability eval-
uation is relevant since the water content of any soil void connects
3.4.3. MICP as a potential source of carbon sequestration: adding directly with stabilization due to the interconnection between
value on unpaved roads effective soil tension (that drives soil resistance) and neutral
Degens et al. (2000), Nannipieri et al. (2003), and Murugan et al. pressure, dependable on promoted tension from water percolation
(2014) studied the role of soil microbial community in the main- (França et al., 2009; Rufino et al., 2011).
tenance of soil ecosystem function, such as C sequestration. Millo Permeability coefficient is the property of the porous media only
et al. (2012) proposed biocementation via ureolytic bacteria as a and is dependent on temperature and porosity (void rate). Higher
8 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

temperature drives to lower water viscosity (), guiding to enhanced pores without subjective assumptions. The author also stated that
percolation rate, increasing the coefficient of permeability (K) the measurement of pore size is a vital step to assess the structure
(Caputo, 1999; França et al., 2009; Rufino et al., 2011). This well- of the soil, directly impacting on how to improve its geotechnical
known factor is commonly neglected on biocementation publica- properties. In our reviews, we note that these assumptions are not
tions since the majority of papers do not state the temperature of often clearly stated, except the porosity values that were calculated,
the environment either of the laboratory tests. This factor alone can measured, or assumed.
drive a test replication to a considerably different result, even The main objective of biocementation is to promote cementa-
misinterpretation of results, which can generate misleading tion between soil particles to increase their stiffness and shear
publications. resistance, maintaining their porosity until the cementation solu-
tion spreads evenly before the completion of the biocementation
3.5.2. Soil stratification & permeability process (Whiffin, 2004; Van Paassen, 2009; Van Paassen et al.,
Complementary to temperature, soil stratification also in- 2010; Van Paassen, 2011; Cheng et al, 2013, 2014). However, a
terferes with permeability rate. Casagrande and Fadum (1940) brief analysis of the porosity concept clarifies the main bottlenecks
described the coefficient of permeability (K) per soil type, were of biocementation: How to maintain a constant and low rate of
coarse material like pure gravel has high permeability, leading to biocementation speed to allow the continuity of soil porosity,
fast percolation rate (10
2 cm/s); and fine particles like clay has low allowing the percolation of the cementation solution by the sample
permeability, reaching impervious level, with almost null percola- without restricting the process of biocementation or compromise
tion rate (10
11 cm/s). Fine sand to sandy soils, including gravel its reinforcement of shear strength? The answer to this question is
mixtures, have a coefficient of permeability between 10
2 and another threshold of the biocementation technique, independent of
10
6 cm/s. Organic and inorganic silts range from 10
5 to 10
9 cm/s, the selected bacteria.
varying from a moderate to a slow percolation rate.
Soil stratification is a relevant constraint on the biocementation 3.5.4. Shear strength
process since most published papers tested biocementation on the Roads are susceptible to constant shear stresses from vehicles
sand and only a few on sandy soils. Whiffin (2004) and Van Paassen and heavy machinery. To guarantee the quality of the road, the
(2009), confirmed by many authors like DeJong et al. (2010), Cheng construction material used needs to meet the minimum require-
and Cord-Ruwisch (2012), Achal and Kawasaki (2016), showed that ment of the shear strength parameter for the planned use (Bakhsh,
in sand or sandy soils, the calcium carbonate precipitate does not 2014). If the material is not homogeneous or does not have shear
have a homogenous distribution over the sample length. Also, the resistance, the road suffers damage, and the collapse is inevitable
biocementation concentrates near the local of cementation in- over time (Machado, 2008). Shear strength is directly related to
jections, since the biocementation process starts during the injec- shear stress and porosity (Terzaghi, 1943; Casal et al., 2001; Vallejo
tion phase, limiting the homogeneous distribution of cementation and Mawby, 2000; Costa et al., 2001).
solution. In theory, and as stated by Van Paassen (2009), soil with a Soil cohesion (c) and friction angle (ɸ) are the main parameters
rapid flow rate facilitates faster percolation of cementation solution that define the shear strength of any material. After extensive
into the sample column, potentially reducing the bioclogging near analysis, we agree with the summary of Mujah et al. (2017) on some
the injection point. This rapid flow is feasible (in theory) because a results of the research by Duraisamy and Airey (2012), Chou et al.
higher percolation rate allows a broader spread of cementation (2011), Ng et al. (2012), Montoya and DeJong (2015), Cheng et al.
solution before the end of the biocementation process, potentially (2013), Soon et al. (2013) and Chu et al. (2012), where they
decreasing the bioclogging. However, van Paassen, Soon, DeJong concluded that the biocemented sandy soils have their cohesion
among other authors tested soils with similar permeability rate and friction angle increased due to the higher concentration of
(sandy soil), and the same bacteria are still searching for the answer CaCO3 (calcite precipitation) filling the spaces porous soil. Nimmo
of the central question: why does biocementation of Sporosarcina (2004) stated that pore size is a crucial component to improve
pasteurii in sandy soil does not reach homogenous distribution the geotechnical properties of the soil since cohesion directly af-
along the sample length, independent of its scale? This answer is fects shear strength and that cohesion is directly affected by pore
another threshold that demands a better understanding that can sizes. The concepts of the shear strength and porosity presented
lead to more predictable outcomes on the biocementation process. here back up our hypothesis that another threshold of the bio-
cementation process depends on understanding the interconnec-
3.5.3. Porosity (void fraction) and pore size distribution tion between shear strength and biocementation rate and how they
Nimmo (2004) and Brady and Weil (2008) define porosity (F) affect each other.
(void fraction) as the ratio between total void volume (occupied by
air or fluid) divided by the total volume of soil, with values between 3.5.5. Soil microstructure
0 and 1, volume or in percentage. Porosity depends on many factors, DeJong et al. (2010) stated that bacterial behavior and the soil
such as packing density, distribution and particle size (polydisperse filtration process determine the spatial distribution of CaCO3 pre-
versus monodisperse), particle shape, and cementation rate (or cipitation. The same authors stated that the bacteria act in particle-
particle welding). Soil with irregular particles tends to form larger to-particle contact due to the lower shear stress and the availability
voids, increasing its porosity. Sandy soils tend to have more of nutrients at a granular level, promoting the concentration of
spherical particles and less cementation, promoting larger voids bacteria in places where resources are abundant. Also, CaCO3
between the particles and a porosity range between 0.30 and 0.35. precipitation reduces porous spaces, replacing the existing fluid
The clay soils, clay, and organic materials have natural cementation (and potential full pore filling), which directly affects the filtration
of the particles, creating large volumes of aggregates, which indi- process, as well as percolation and permeability rates. This pore
vidually support a porosity of 0.35, but as a soil profile, they have a space reduction forces the precipitated CaCO3 to stick close to soil
porosity greater than 0.5. Soils with a high organic matter content particles as the porous fluid flows through the pore throat, reducing
can reach very low porosity of 0.8e0.9. However, Nimmo (2004) the pore space and potentially bioclogging (pore-clogging due to
emphasizes that the soil-water-air system has an intrinsic CaCO3 deposition) (DeJong et al., 2006).
behavior and soil particles do not have a unique shape or size, The soil microstructure is another constraint for a successful
making it almost impossible to delineate the size and shape of the biocementation process, regardless of the type of soil or the
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 9

selected bacteria. Scanning electron microscopy (SEM) images are stabilization by cement or lime is the most used stabilization
the tools to evaluate the biocementation process, assessing the technique and promotes alteration in bearing capacity, shear
amount and distribution of CaCO3 bonds with soil particles, allied strength and permeability (Teng et al., 2007; Ismail et al., 2002;
with resistance tests to assess the stability of biocemented soil Machado, 2008; Bakhsh, 2014), culminating in high stiffness and
(Whiffin, 2004). DeJong et al. (2010) found that only precipitated brittle behavior (Wang et al., 2003; Basha et al., 2005; Bakhsh,
CaCO3 crystals that form an effective bond between sand particles 2014). A high stiffness soil means soil with the high elastic
contribute to increasing the shear strength of biocemented sand. modulus (E), defined by a high ratio of stress over strain, translating
These calcite-particle bonds in the soil occur at the level of inter- the bonding strength into loose soil grains (Mujah et al., 2017).
particle, replacing the fluid and must have a uniform distribution to Yang and Gu (2013) stated that confining stress and void ratio
promote soil resistance. However, CaCO3 precipitation tends to are the main factors affecting shear stiffness (G0). They cited the
follow a unique pattern of distribution, concentrating in the vicinity Wichtmann and Triantafyllidis (2009) research with sand, which
of the bacteria’s nucleation site. confirmed that the shear stiffness (G0) decreases considerably as
the coefficient of uniformity (Cu) increases. This ratio means that
3.6. MICP on unsealed roads soil with a higher concentration of particles within uniform sizes
leads to a natural decrease in shear stiffness. For this same reason,
Unsealed roads are necessary for the development of any region chemical stabilization is the method to reduce the erosion of sub-
since they provide social-economic and accessibility benefits grades of pavements (Machado et al., 2009). In Brazil, unsealed
(Rammelt and Leung, 2017), and have a much lower cost than paved roads can not use sand or sandy soils, and the soil must be gran-
roads. Unsealed roads in Brazil (a similar situation for the majority ulometric stabilized, which contains a low volume of sand (IPR/
of the developing countries) used for temporary purposes such as DNIT, 2006). Granulometric stabilized soils have an increased
mining industry access, and as permanent roads in rural areas, shear stiffness. However, this enhanced shear stiffness is not strong
reach a considerable percentage of total constructed roads enough to support traffic, which demands extra stabilization, often
(Ministerio de Obras Publicas, 2018). The majority of these unsealed chemical stabilization with cement or lime (Machado et al., 2009;
roads have massive traffic demand, which increases the demand for Bakhsh, 2014). Lee et al. (2013) tested biocementation on residual
maintenance to sustain the performance and security of the roads. soil (43% of silt, 38% of sand, 19% of clay, and 0% of gravel) and found
Besides, many soils are not suitable to carry loads (be used as road a similar stiffness behavior than on biocemented natural sand.
subgrade) without a high cost of maintenance in the long-term Rebata-Landa (2007) suggested a grain size ranging between 50
(Cabezas and Cataldo, 2019). and 400 mm to attend the bacterial activity requirement of a low
Stabilization methods applied to in-situ soils are necessary for concentration of very fine soil particles (clay). Also, coarse materials
improving mechanical behavior via improving bearing capacity or demand high production of calcite to fulfill the high number of
decreasing permeability (or both) and reducing maintenance costs voids, biocementing the soil particles without compromise the
over time (Parsons and Milburn, 2003). The standards for unsealed stiffness. This review shows that any study about soil stabilization
road construction in the US and Brazil demands granulometric and unsealed road construction using biocementation procedure
stabilization of the soil to improve its stability to support the should include soil stiffness as it is a critical limit factor.
designed road. In countries like Brazil, where only 213,453 km
(12.4%) from the 1.720 million kilometers of the total road network
has asphalt or similar coverage, chemical stabilization is widely
used (CNT, 2018). Biocementation presents itself as a potential 3.6.3. Erosion on unsealed roads: chemical stabilization methods
green alternative since it has the potential to substitute the use of and MICP
cement or lime on road construction in Brazil. The erosion of unsealed roads begins when the runoff is
concentrated along the road drainage channel, staggering the shear
3.6.1. Granulometric stabilized soil - the foundation for unsealed stress until it surpasses the critical shear stress of the road surface.
road construction From this moment on, the runoff will carry out the soil particles,
Granulometric stabilization is the alteration of soil properties contributing to the local watershed sedimentation process. The
through the mixture of different sizes and concentrations of gravel, well-described impacts include sedimentation, water quality
sand, silt, and clay particles resulting in a homogenized mixture degradation, which directly affects the aquatic life of watercourses
followed by compaction under Proctor energy parameters (Ziegler et al., 2000; 2001; Zhang et al., 2009; Liu et al., 2009;
(Vizcarra, 2010). Machado et al. (2009) adds that granulometric Correa and Cruz, 2010, Bakhsh, 2014).
stabilization of road subgrade focuses on a final composite of ma- Terzaghi (1943) and Terzaghi et al. (1996) proved that shear
terials that attend the designed road cargo demand. IPR/DNIT stress in soil mechanics is the main force responsible for ruptures in
(2006) and IPR/DNIT (2010) determine that the granulometric slopes, valleys, dams, and other geomechanical forces over a sedi-
distribution is the percentage of each particle size present on soil mentary soil. The authors also stated that clay soils demand
mixture and its distribution. Then, the granulometric soil mixture exhaustive analysis and studies of its mechanics based on intended
receives a mechanical stabilization through Proctor compaction, use, since micro-clays (or clay-minerals) have a larger buffer of
finalizing the granulometric stabilization of the soil. This stage is water around the particles, meaning that the fluctuation of the
the granulometric stabilized soil or unsealed road, which is also the water content of soil directly affects road stabilization. Citing
subgrade for the paved roads. Even after granulometric stabiliza- Umesh et al. (2011), Vakili et al. (2013) and Edgar (1991),
tion, unsealed roads are highly susceptible to erosion processes, Premkumar et al. (2016) stated that Brazil and the United States are
especially hydric erosion. For this reason, the Brazilian unsealed two of the top nine countries that have dispersive soils, meaning
road standards (and also the US) promote the additional chemical that the clay particles are dispersible and behave as a single-
stabilization methods, like the classic addition of cement or lime. grained particle. Premkumar et al. (2016), citing Sherard and
Decker (1977); Bell (2003); Biggs and Mahony (2004), summa-
3.6.2. Soils stiffness and stabilization rized that dispersive soils lose the interparticle forces and suffer
As stated at the 3.4.1 subtopic, chemical stabilization of the soil from erosion failure when exposed to water, meaning that these
is a requirement from road construction standards. Chemical soils have low wet bearing strength.
10 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

3.6.4. Natural calcite precipitation in soils - a prediction model Summarizing the papers evaluated in this review, the ideal bio-
The soil and agricultural sciences established that calcite pre- cementation model should consider three central factors, at least.
cipitation occurs naturally near the root surface once the root First, guarantee a uniform and balanced environment for bacteria,
quickly absorbs nitrate and release bicarbonate to the soil to including the bacteria-specific process, to calculate the necessary
maintain the charge balance at the root-soil boundary. This pre- concentration of the bacteria. Second, define the relevant porosity
cipitation led to a pH increase, usually enough to facilitate natural combined with the correct concentration of the cementation so-
CaCO3 precipitation near the root surface (Nye, 1981; Kirk and Nye, lution to ensure homogeneity of the CaCO3 precipitation. Third, use
1991; Neumann and Ro € mheld, 2012). This process called the first and second factors to eliminate the need for multiple in-
microbially-enhanced urea hydrolysis is also known as MICP, hav- jection points of the cementation solution or successive applica-
ing Stocks-Fischer et al. (1999) as one of the first research to pro- tions in the same location, thus reducing operating costs.
pose its use to stabilize soils. Urea hydrolysis processes driven by Morales et al. (2015) research sustain our rationale that it is
microbial enhanced or induced calcite precipitation have some necessary to develop an optimum biocementation solution that
points in common: reactants promote local CaCO3 precipitation; must be available during the MICP process to facilitate the CaCO3
the rate and distribution of precipitation are directly dependent on precipitation. This rationale also applies to the research aiming to
the in-out reactant transportation rates at the precipitation zone, promote a sustainable unpaved road biostabilization. Since no
and mainly by precipitation kinetics at the nucleation sites per se. research has developed to elucidate the optimum concentration of
Most studies reviewed in this article do not state clear information bacteria, bacteria species selection (indigenous species to minimize
about transport and precipitation kinetics or do not even mention it potential hazardous impact and cost and with a high rate of CaCo3
at all. Kirk et al. (2015) summarize three significant factors that can precipitation), cementation solutions and soil mixture, our primary
affect transport and precipitation kinetics. First, the CaCO3 kinetics research continues to pursue the goal of developing an experi-
in simple solutions is well described but still lack information about mental study to develop this innovative field further.
its interaction with soil systems and other porous media. Second, it In summary, this review showed that MICP is feasible as a soil
is common to have soil solutions supersaturated with CaCO3, which stabilization process but still needs considerable development. The
directly affects precipitation due to its sensitive reaction to CaCO3 protocol development should anchor on a considerable break-
and other fluids catalysis and inhibition by organic and inorganic through that addresses the intricacies of MICP stages and its six
ligands present at the soil solution. Third, the fact that the soil main limiting factors: calcium concentration, dissolved inorganic
transport rate usually is lower than simple solution systems due to carbon (DIC) concentration, pH of the media, availability of nucle-
most solutes being sorbed on soil surface becoming mostly ation sites, urease activity; and carbonic anhydrase activity. More
immobile in the sorbed state. exploratory studies are necessary to evaluate the behavior of each
Kirk et al. (2015) proposed a simple reactive-transport model to proposed bacterial isolates facing the limiting factors and the
evaluate the calcite precipitation in soils and concluded that it is improvement of the mechanical properties of granular soils
vital to evaluate the initial soil pH, the soil pH buffer power, and the (including unsealed roads). This review also delineated another gap
CO2 pressure to promote a long-spread zone of precipitation from for any MICP soil stabilization: which is the best injection method
its origin. It is necessary for a small pH buffer power for sandy soils to achieve a homogenous biocementation for the proposed use of
or sub-strata and desired a high CO2 pressure for clayey soils to the soil?
sustain a high biological activity. The model proved the importance
of the geometric system and the spacing between macro-pores
where the precipitation is happening. By analyzing the model, we Funding
can extend the following question to any biocementation process:
How to predict and guarantee the desired conditions of soil pH, soil Funding for this research includes scholarship (Process n.: 8453/
pH buffer, CO2 pressure, reactant transport rate, and its movement 13-6) and financial funding (Process n.: 472393-2013/8) from the
of in-and-out precipitation zone to promote homogenous CaCO3 Coordenaça ~o de Aperfeiçoamento de Pessoal de Nível Superior
precipitation? (CAPES, Coordination for the Improvement of Higher Education
Personnel, a Brazilian federal government agency under the Min-
4. Conclusions & considerations istry of Education), Brazil; the Department of Forest Engineering
and Department of Microbiology of the Federal University of Viçosa,
Studies by Soon et al. (2013) have shown the effectiveness of Brazil; the BioAgro e Instituto de Biotecnologia Aplicada a  Agro-
MICP by an improvement of 96% on soil shear strength and reduced pecuaria (Institute of Biotechnology Applied Agriculture), Brazil;
soil permeability. The successful research on MICP for soil stabili- and the Oregon State University e Environmental Sciences Grad-
zation should focus on: the identification of microorganisms with uate Program, USA.
higher calcium carbonate precipitation; soil optimal conditions to
increase the microorganism growth rate (ureolytic activity); anal-
ysis of the calcium carbonate production rate under field condi- Declaration of competing interest
tions; and on differences between indigenous and exotic
microorganism growth rates (Whiffin et al., 2007; Van Paassen, The authors declare that they have no known competing
2011; Cheng and Cord-Ruwisch, 2012; Yasuhara et al., 2011; financial interests or personal relationships that could have
Shahrokhi-Shahraki et al., 2015). Field studies of soil bio- appeared to influence the work reported in this paper.
stabilization at a large scale are still scarce given the complexity of
the process, the long-term research cycle, and the high cost of the
field-scale research projects. However, the personal/professional Acknowledgments
desire to publish before any other researcher has catalyzed several
scientists to repeat very similar experiments and rush publication The authors acknowledge the valuable support from the Aca-
in this field. Studies and tests are necessary to obtain the best re- demic Writing Center at Oregon State University, Rodrigo, Ligia,
sults of the MICP for each environment and road conditions to Angelica, and Raiane for providing writing assistance, language
continue to improve field techniques and applicable outcomes. support, and proofreading the article.
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 11

References Chou, C.W., Seagren, E.A., Aydilek, A.H., Lai, M., 2011. Biocalcification of sand
through ureolysis. J. Geotech. Geoenviron. Eng. 137 (12), 1179e1189.
Chu, J., Ivanov, V., Stabnikov, V., He, J., Li, B., Naemi, M., 2012. Biocement: green
Achal, V., Kawasaki, S., 2016. Biogrout: a novel binding material for soil improve-
building-and energy-saving material. In: Advanced Materials Research, vol. 347.
ment and concrete repair. Front. Microbiol. 7, 314.
Trans Tech Publications Ltd, pp. 4051e4054.
Achal, V., Pan, X., 2011. Characterization of urease and carbonic anhydrase pro-
CNT, 2018. Pesquisa CNT de rodovias 2018: relato  rio gerencial. Confederaç~ ao
ducing bacteria and their role in calcite precipitation. Curr. Microbiol. 62 (3),
Nacional do Transporte, SEST, Serviço Social do Transporte, SENAT, Serviço
894e902.
Nacional de Aprendizagem do Transporte (Brasília, Brazil).
Achal, V., Mukherjee, A., Basu, P.C., Reddy, M.S., 2009a. Lactose mother liquor as an
Coleman-Derr, D., Desgarennes, D., Fonseca-Garcia, C., Gross, S., Clingenpeel, S.,
alternative nutrient source for microbial concrete production by Sporosarcina
Woyke, T., et al., 2016. Plant compartment and biogeography affect microbiome
pasteurii. J. Ind. Microbiol. Biotechnol. 36 (3), 433e438.
composition in cultivated and native Agave species. New Phytol. 209 (2),
Achal, V., Mukherjee, A., Basu, P.C., Reddy, M.S., 2009b. Strain improvement of
798e811.
Sporosarcina pasteurii for enhanced urease and calcite production. J. Ind.
Correa, C.M.C., Cruz, J., 2010. Eros~ ao real e estimada atrave s da RUSLE em estradas
Microbiol. Biotechnol. 36 (7), 981e988. 
florestais, em relevo ondulado a fortemente ondulado. Rev. Arvore 34 (4),
Achal, V., Mukherjee, A., Kumari, D., Zhang, Q., 2015. Biomineralization for sus-
587e595.
tainable constructioneA review of processes and applications. Earth Sci. Rev.
Costa, M.L., De Almeida, S.F.M., Rezende, M.C., 2001. The influence of porosity on the
148, 1e17.
interlaminar shear strength of carbon/epoxy and carbon/bismaleimide fabric
Al Qabany, A., Soga, K., Santamarina, C., 2012. Factors affecting efficiency of
laminates. Compos. Sci. Technol. 61 (14), 2101e2108.
microbially induced calcite precipitation. J. Geotech. Geoenviron. Eng. 138 (8),
Cunningham, A.B., Phillips, A.J., Troyer, E., Lauchnor, R., Hiebert, R., Gerlach, R.,
992e1001.
Spangler, L.H., 2014. Wellbore leakage mitigation using engineered biominer-
Al-Thawadi, S., 2008. High Strength In-Situ Biocementation of Soil by Calcite
alization. Energy Procedia 63, 4612e4619.
Precipitating Locally Isolated Ureolytic Bacteria. Doctoral dissertation. Murdoch
De Muynck, W., De Belie, N., Verstraete, W., 2010. Microbial carbonate precipitation
University.
in construction materials: a review. Ecol. Eng. 36 (2), 118e136.
Aloisi, G., Gloter, A., Kruger, M., Wallmann, K., Guyot, F., Zuddas, P., 2006. Nucleation
Degens, B.P., Schipper, L.A., Sparling, G.P., Vojvodic-Vukovic, M., 2000. Decreases in
of calcium carbonate on bacterial nanoglobules. Geology 34 (12), 1017e1020.
organic C reserves in soils can reduce the catabolic diversity of soil microbial
Anbu, P., Kang, C.H., Shin, Y.J., So, J.S., 2016. Formations of calcium carbonate min-
communities. Soil Biol. Biochem. 32 (2), 189e196.
erals by bacteria and its multiple applications. SpringerPlus 5 (1), 1e26.
DeJong, J.T., Fritzges, M.B., Nüsslein, K., 2006. Microbially induced cementation to
Bakhsh, K.N., 2014. Design Methodology for Subgrades and Bases under Concrete
control sand response to undrained shear. J. Geotech. Geoenviron. Eng. 132 (11),
Roads and Parking Lots. PhD diss.. Texas A&M University, College Station, TX.
1381e1392.
Banks, E.D., Taylor, N.M., Gulley, J., Lubbers, B.R., Giarrizzo, J.G., Bullen, H.A., et al.,
DeJong, J.T., Mortensen, B.M., Martinez, B.C., Nelson, D.C., 2010. Bio-mediated soil
2010. Bacterial calcium carbonate precipitation in cave environments: a func-
improvement. Ecol. Eng. 36 (2), 197e210.
tion of calcium homeostasis. Geomicrobiol. J. 27 (5), 444e454.
DeJong, J.T., Soga, K., Banwart, S.A., Whalley, W.R., Ginn, T.R., Nelson, D.C., et al.,
Basha, E.A., Hashim, R., Mahmud, H.B., Muntohar, A.S., 2005. Stabilization of re-
2011. Soil engineering in vivo: harnessing natural biogeochemical systems for
sidual soil with rice husk ash and cement. Construct. Build. Mater. 19 (6),
sustainable, multi-functional engineering solutions. J. R. Soc. Interface 8 (54),
448e453.
1e15.
Bell, F.G., 2003. Geological hazards: their assessment, avoidance and mitigation. CRC
DeJong, J.T., Soga, K., Kavazanjian, E., Burns, S., Van Paassen, L.A., Al Qabany, A., et al.,
Press.
lcio: uma 2013. Biogeochemical processes and geotechnical applications: progress, op-
Bessler, K.E., Rodrigues, L.C., 2008. Os polimorfos de carbonato de ca
portunities and challenges. Geotechnique 63 (4), 287e301.
síntese facil de aragonita. Quím. Nova 31 (1), 178e180.
Dhami, N.K., Sudhakara, R.M., Abhijit, M., 2013. Biomineralization of calcium car-
Bibi, S., Oualha, M., Ashfaq, M.Y., Suleiman, M.T., Zouari, N., 2018. Isolation, differ-
bonates and their engineered applications: a review. Front. Microbiol. 4 (314),
entiation and biodiversity of ureolytic bacteria of Qatari soil and their potential
1e13. https://doi.org/10.3389/fmicb.2013.00314.
in microbially induced calcite precipitation (MICP) for soil stabilization. RSC
Dhami, N.K., Reddy, M.S., Mukherjee, A., 2014. Application of calcifying bacteria for
Adv. 8 (11), 5854e5863.
remediation of stones and cultural heritages. Front. Microbiol. 5, 304.
Biggs, A., Mahony, K., 2004. Is soil science relevant to road infrastructure?. In: Proc.,
Dhami, N.K., Mukherjee, A., Reddy, M.S., 2016. Applicability of bacterial bio-
13th Int. Soil Conservation Organization Conf., (ISCO)dConserving Soil and
cementation in sustainable construction materials. Asia Pac. J. Chem. Eng. 11 (5),
Water for Society: Sharing Solutions, ISCO, Tucson, Brisbane, Australia.
795e802.
Bontognali, T.R., Vasconcelos, C., Warthmann, R.J., Dupraz, C., Bernasconi, S.M.,
Donnelly, F.C., Purcell-Milton, F., Framont, V., Cleary, O., Dunne, P.W., Gun’ko, Y.K.,
McKenzie, J.A., 2008. Microbes produce nanobacteria-like structures, avoiding
2017. Synthesis of CaCO3 nano-and micro-particles by dry ice carbonation.
cell entombment. Geology 36 (8), 663e666.
Chem. Commun. 53 (49), 6657e6660.
Bosak, T., Newman, D.K., 2003. Microbial nucleation of calcium carbonate in the
Dupraz, S., Parmentier, M., Me nez, B., Guyot, F., 2009a. Experimental and numerical
Precambrian. Geology 31 (7), 577e580.
modeling of bacterially induced pH increase and calcite precipitation in saline
Boskey, A.L., 1998. Biomineralization: conflicts, challenges, and opportunities. J. Cell.
aquifers. Chem. Geol. 265 (1e2), 44e53.
Biochem. 72 (S30‒31), 83e91.
Dupraz, S., Me nez, B., Gouze, P., Leprovost, R., Bene
zeth, P., Pokrovsky, O.S., Guyot, F.,
Brady, N.C., Weil, R.R., 2008. The soils around us. In: The Nature and Properties of
2009b. Experimental approach of CO2 biomineralization in deep saline aquifers.
Soils, fourteenth ed. Pearson Prentice Hall, New Jersey and Ohio, pp. 1e31.
Chem. Geol. 265 (1e2), 54e62.
Cabezas, R., Cataldo, C., 2019. Influence of chemical stabilization method and its
Duraisamy, Y., Airey, D.W., 2012. Strength and stiffness of bio-cemented liquefiable
effective additive concentration (EAC) in non-pavement roads: a study in
sand soil. In: Proceedings of the International Conference on Ground
andesite-based soils. Cogent Eng. 6 (1), 1592658.
Improvement and Ground Control, Singapore, pp. 1233e1239.
Cacchio, P., Ercole, C., Cappuccio, G., Lepidi, A., 2003. Calcium carbonate precipita-
Edgar, H.N., 1991. “Soil Mechanics Note No. 13: Dispersive clays.” Soil Conservation
tion by bacterial strains isolated from a limestone cave and from a loamy soil.
Service. United States Dept. of Agriculture, Washington, DC, pp. 11e12.
Geomicrobiol. J. 20 (2), 85e98.
Enriquez, A.G., 2017. Biocimentaç~ ao de Solos Arenosos para fins de utilizaç~ ao em
Caputo, M., 1999. Diffusion of fluids in porous media with memory. Geothermics 28
Estradas Na ~o Pavimentadas. Doctoral dissertation, PhD Thesis. Universidade
(1), 113e130.
Federal de Viçosa, Viçosa, Minas Gerais, Brasil.
Casagrande, A., Fadum, R.E., 1940. Notes on soil testing for engineering purposes. In:
Ferna ndez, M.S., Montt, B., Ortiz, L., Neira-Carrillo, A., Arias, J.L., 2018. Effect of
Soil Mechanics Series, vol. 8. Graduate School of Engineering, Harvard Univer-
carbonic anhydrase immobilized on eggshell membranes on calcium carbonate
sity, Cambridge, Massachusetts.
ndez, R., 2001. Influence of crystallization in vitro. In Biomineralization 31e37. Springer, Singapore.
Casal, E., Granda, M., Bermejo, J., Bonhomme, J., Mene
Ferris, F.G., Phoenix, V., Fujita, Y., Smith, R.W., 2004. Kinetics of calcite precipitation
porosity on the apparent interlaminar shear strength of pitch-based unidirec-
induced by ureolytic bacteria at 10 to 20 C in artificial groundwater. Geochem.
tional CeC composites. Carbon 39 (1), 73e82.
tayer-Levrel, G., Perthuisot, J.P., 1999. Ca-carbonates precipitation Cosmochim. Acta 68 (8), 1701e1710.
Castanier, S., Le Me
Frankel, R.B., Bazylinski, D.A., 2003. Biologically induced mineralization by bacteria.
and limestone genesisdthe microbiogeologist point of view. Sediment. Geol.
Rev. Mineral. Geochem. 54 (1), 95e114.
126 (1e4), 9e23.
 pez, C.E., Narayanasamy, R., Marszalek, J.E., Luevanos-Escaren ~ o, M.P., França, A.C., Carrocci, L.R., Siqueira, A.F., 2009. Velocity profile visualization of water
Castro, M.J., Lo
natural percolation in a porous medium. Rev. Engenharia Te rmica 8 (1), 31e35.
Fajardo, G.J., Balagurusamy, N., 2016. Potential of enzymes (urease & carbonic
Fritzges, M., 2005. Biologically Induced Improvements of the Response of Sands to
anhydrase). Chim. Oggi-Chem. Today 34, 4.
Monotonic Loading. M.S. thesis. Univ. of Massachusetts, Amherst, MA.
Chapelle, F.H., 2000. Ground-water Microbiology and Geochemistry. John Wiley &
Fujita, Y., Taylor, J.L., Gresham, T.L., Delwiche, M.E., Colwell, F.S., McLing, T.L., et al.,
Sons.
2008. Stimulation of microbial urea hydrolysis in groundwater to enhance
Cheng, L., Cord-Ruwisch, R., 2012. In situ soil cementation with ureolytic bacteria by
calcite precipitation. Environ. Sci. Technol. 42 (8), 3025e3032.
surface percolation. Ecol. Eng. 42, 64e72.
Fukue, M., Sato, Y., Yamashita, M., Yanai, M., Fujimori, Y., 2003. Change in micro-
Cheng, L., Cord-Ruwisch, R., Shahin, M.A., 2013. Cementation of sand soil by
structure of soils due to natural mineralization. Appl. Clay Sci. 23 (1e4),
microbially induced calcite precipitation at various degrees of saturation. Can.
169e177.
Geotech. J. 50 (1), 81e90.
Gao, Y., Hang, L., He, J., Chu, J., 2019. Mechanical behavior of biocemented sands at
Cheng, L., Shahin, M.A., Cord-Ruwisch, R., 2014. Bio-cementation of sandy soil using
various treatment levels and relative densities. Acta Geotech. 14 (3), 697e707.
microbially induced carbonate precipitation for marine environments. Geo-
Gat, D., Tsesarsky, M., Shamir, D., Ronen, Z., 2014. Accelerated microbial-induced
technique 64 (12), 1010e1013.
CaCO3 precipitation in a defined coculture of ureolytic and non-ureolytic
12 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372

bacteria. Biogeosciences 11 (10), 2561. solubility-trapping. Environ. Sci. Technol. 44 (13), 5270e5276.
Hammes, F., Verstraete, W., 2002. Key roles of pH and calcium metabolism in mi- Mitchell, A.C., Ferris, F.G., 2005. The coprecipitation of Sr into calcite precipitates
crobial carbonate precipitation. Rev. Environ. Sci. Biotechnol. 1 (1), 3e7. induced by bacterial ureolysis in artificial groundwater: temperature and ki-
Hammes, F., Boon, N., de Villiers, J., Verstraete, W., Siciliano, S.D., 2003. Strain- netic dependence. Geochem. Cosmochim. Acta 69 (17), 4199e4210.
specific ureolytic microbial calcium carbonate precipitation. Appl. Environ. Mitchell, J.K., Santamarina, J.C., 2005. Biological considerations in geotechnical
Microbiol. 69 (8), 4901e4909. engineering. J. Geotech. Geoenviron. Eng. 131 (10), 1222e1233.
Handley-Sidhu, S., Sham, E., Cuthbert, M.O., Nougarol, S., Mantle, M., Johns, M.L., Mitchell, A.C., Phillips, A., Schultz, L., Parks, S., Spangler, L., Cunningham, A.B.,
et al., 2013. Kinetics of urease mediated calcite precipitation and permeability Gerlach, R., 2013. Microbial CaCO3 mineral formation and stability in an
reduction of porous media evidenced by magnetic resonance imaging. Int. J. experimentally simulated high pressure saline aquifer with supercritical CO2.
Environ. Sci. Technol. 10 (5), 881e890. Int. J. Greenh. Gas Control 15, 86e96.
Harkes, M.P., Van Paassen, L.A., Booster, J.L., Whiffin, V.S., van Loosdrecht, M.C., 2010. Montoya, B.M., DeJong, J.T., 2015. Stress-strain behavior of sands cemented by
Fixation and distribution of bacterial activity in sand to induce carbonate pre- microbially induced calcite precipitation. J. Geotech. Geoenviron. Eng. 141 (6),
cipitation for ground reinforcement. Ecol. Eng. 36 (2), 112e117. 04015019.
He, J., Gao, Y., Gu, Z., Chu, J., Wang, L., 2020. Characterization of crude bacterial Morales, L., Romero, E., Jommi, C., Garzo n, E., Gime nez, A., 2015. Feasibility of a soft
urease for CaCO3 precipitation and cementation of silty sand. J. Mater. Civ. Eng. biological improvement of natural soils used in compacted linear earth con-
32 (5), 04020071. struction. Acta Geotech. 10 (1), 157e171.
Hokkanen, H.M., Lynch, J.M., 2003. Biological control: benefits and risks, vol. 4. Mujah, D., Shahin, M.A., Cheng, L., 2017. State-of-the-art review of biocementation
Cambridge University Press. by microbially induced calcite precipitation (MICP) for soil stabilization. Geo-
Hwang, E.T., Gang, H., Gu, M.B., 2013. CO2 bioconversion using carbonic anhydrase microbiol. J. 34 (6), 524e537.
(CA): effects of PEG rigidity on the structure of bio-mineralized crystal com- Müller, W.E., Neufurth, M., Schlossmacher, U., Schro €der, H.C., Pisignano, D.,
posites. J. Biotechnol. 168 (2), 208e211. Wang, X., 2014. The sponge silicatein-interacting protein silintaphin-2 blocks
IPR/DNIT, 2006. Manual de pavimentaça ~o. In: Instituto de Pesquisas Rodovia rias e calcite formation of calcareous sponge spicules at the vaterite stage. RSC Adv. 4
Departamento Nacional de Infraestrutura de Transportes. Diretoria de Plane- (6), 2577e2585.
jamento e Pesquisa. Coordenaça ~o Geral de estudos e pesquisa. IPR Publ. 719, Murugan, R., Loges, R., Taube, F., Sradnick, A., Joergensen, R.G., 2014. Changes in soil
p. 274. microbial biomass and residual indices as ecological indicators of land use
IPR/DNIT, 2010. Norma DNIT 141/2010-ES: pavimentaça ~o - base estabilizada gran- change in temperate permanent grassland. Microb. Ecol. 67 (4), 907e918.
ulometricamente - especificaça ~o de serviço. In: Instituto de Pesquisas Rodo- Nannipieri, P., Ascher, J., Ceccherini, M., Landi, L., Pietramellara, G., Renella, G., 2003.
rias e Departamento Nacional de Infraestrutura de Transportes. Diretoria de
via Microbial diversity and soil functions. Eur. J. Soil Sci. 54 (4), 655e670.
Planejamento e Pesquisa. Coordenaç~ ao Geral de estudos e pesquisa, p. 10. Neumann, G., Ro €mheld, V., 2012. Rhizosphere chemistry in relation to plant
Ismail, M.A., Joer, H.A., Sim, W.H., Randolph, M.F., 2002. Effect of cement type on nutrition. In: Marschner’s mineral nutrition of higher plants. Academic Press,
shear behavior of cemented calcareous soil. J. Geotech. Geoenviron. Eng. 128 (6), pp. 347e368.
520e529. Ng, W.S., Lee, M.L., Hii, S.L., 2012. An overview of the factors affecting microbial-
Ivanov, Volodymyr, 2010. Environmental Microbiology for Engineers. CRC Press. induced calcite precipitation and its potential application in soil improve-
Ivanov, V., Chu, J., 2008. Applications of microorganisms to geotechnical engi- ment. World Acad. Sci. Eng. Technol. 62 (2), 723e729.
neering for bioclogging and biocementation of soil in situ. Rev. Environ. Sci. Nimmo, J.R., 2004. Porosity and pore size distribution. Encycl. Soils Environ. 3 (1),
Biotechnol. 7 (2), 139e153. 295e303.
Jaji, A.Z., Zakaria, Z.A.B., Mahmud, R., Loqman, M.Y., Hezmee, M.N.M., Abba, Y., et al., Nye, P.H., 1981. Changes of pH across the rhizosphere induced by roots. Plant Soil 61
2017. Safety assessments of subcutaneous doses of aragonite calcium carbonate (1e2), 7e26.
nanocrystals in rats. J. Nanoparticle Res. 19 (5), 175. Obst, M., Dynes, J.J., Lawrence, J.R., Swerhone, G.D.W., Benzerara, K.,
Jime nez-Lo  pez, C., Rodriguez-Navarro, C., Pin ~ ar, G., Carrillo-Rosúa, F.J., Rodriguez- Karunakaran, C., et al., 2009. Precipitation of amorphous CaCO3 (aragonite-like)
Gallego, M., Gonzalez-Mun ~ oz, M.T., 2007. Consolidation of degraded orna- by cyanobacteria: a STXM study of the influence of EPS on the nucleation
mental porous limestone stone by calcium carbonate precipitation induced by process. Geochem. Cosmochim. Acta 73 (14), 4180e4198.
the microbiota inhabiting the stone. Chemosphere 68 (10), 1929e1936. Osinubi, K.J., Eberemu, A.O., Ijimdiya, T.S., Yakubu, S.E., Gadzama, E.W., Sani, J.E.,
Jonkers, H.M., Thijssen, A., Muyzer, G., Copuroglu, O., Schlangen, E., 2010. Applica- Yohanna, P., 2020. Review of the use of microorganisms in geotechnical engi-
tion of bacteria as self-healing agent for the development of sustainable con- neering applications. SN Appl. Sci. 2 (2), 1e19.
crete. Ecol. Eng. 36 (2), 230e235. €
Ozen, _ Şimşek, S., 2015. Vital importance of moisture level in all stages of pro-
I.,
Kirk, G.J.D., Nye, P.H., 1991. A model of ammonia volatilization from applied urea. V. cessing from calcium carbonate coating through polyethylene/calcium car-
The effects of steady-state drainage and evaporation. J. Soil Sci. 42 (1), 103e113. bonate compounding to film generation. Powder Technol. 270, 320e328.
Kirk, G.J., Versteegen, A., Ritz, K., Milodowski, A.E., 2015. A simple reactive-transport Parsons, R.L., Milburn, J.P., 2003. Engineering behavior of stabilized soils. Transport.
model of calcite precipitation in soils and other porous media. Geochem. Cos- Res. Rec. 1837 (1), 20e29.
mochim. Acta 165, 108e122. Phillips, A.J., Gerlach, R., Lauchnor, E., Mitchell, A.C., Cunningham, A.B., Spangler, L.,
Kucharski, E.S., Cord-Ruwisch, R., Whiffin, V.S., Al-Thawadi, S.M.J., 2006. Microbial 2013. Engineered applications of ureolytic biomineralization: a review.
Biocementation. World Patent WO/2006/066326, June 29. Biofouling 29 (6), 715e733.
Kucharski, E.S., Cord-Ruwisch, R., Whiffin, V., Al-Thawadi, S.M., 2012. U.S. Patent No. Phillips, A.J., Cunningham, A.B., Gerlach, R., Hiebert, R., Hwang, C., Lomans, B.P.,
8,182,604. U.S. Patent and Trademark Office, Washington, DC. et al., 2016. Fracture sealing with microbially-induced calcium carbonate pre-
Lee, M.L., Ng, W.S., Tanaka, Y., 2013. Stress-deformation and compressibility re- cipitation: a field study. Environ. Sci. Technol. 50 (7), 4111e4117.
sponses of bio-mediated residual soils. Ecol. Eng. 60, 142e149. Piekarska, K., Piorkowska, E., Bojda, J., 2017. The influence of matrix crystallinity,
Li, M., Cheng, X., Guo, H., 2013. Heavy metal removal by biomineralization of urease filler grain size and modification on properties of PLA/calcium carbonate
producing bacteria isolated from soil. Int. Biodeterior. Biodegrad. 76, 81e85. composites. Polym. Test. 62, 203e209.
Liu, J., Li, Y., Zhang, B., Cao, J., Cao, Z., Domagalski, J., 2009. Ecological risk of heavy Premkumar, S., Piratheepan, J., Arulrajah, A., Disfani, M.M., Rajeev, P., 2016. Exper-
metals in sediments of the Luan River source water. Ecotoxicology 18 (6), imental study on contact erosion failure in pavement embankment with
748e758. dispersive clay. J. Mater. Civ. Eng. 28 (4), 04015179.
Lowenstam, H.A., Weiner, S., 1989. On Biomineralization. Oxford University Press on Rammelt, C.F., Leung, M.W., 2017. Tracing the causal loops through local perceptions
Demand. of rural road impacts in Ethiopia. World Dev. 95, 1e14.
Machado, C.C., 2008. Manual de construç~ ao e manutença ~o de estradas florestais Rebata-Landa, V., 2007. Microbial Activity in Sediments: Effects on Soil Behavior.
ambientalmente corretas. COPENER-Bahia Pulp. Viçosa, MG 1 & 2. Doctoral dissertation. Georgia Institute of Technology.
Machado, C.C., Santanna, G.L., Lima, D.C.D., Carvalho, C.A.B.D., Pereira, R.S., Ronholm, J., Schumann, D., Sapers, H.M., Izawa, M., Applin, D., Berg, B., et al., 2014.
Fernandes, D.C.D.M., 2009. Comportamento geote cnico de misturas gran- A mineralogical characterization of biogenic calcium carbonates precipitated by
ulome 
tricas de solo-grits. Rev. Arvore 33 (3), 555e562. heterotrophic bacteria isolated from cryophilic polar regions. Geobiology 12 (6),
Madigan, M., 2012. Biodata of Dr. Michael T. Madigan author of “Bacterial Habitats’ 542e556.
in Extreme Environments”. Journey to Diverse Microbial Worlds 2, 61. Rufino, R.D., Rodrigues, G.I.B., Campos-Takaki, G.M., Sarubbo, L.A., Ferreira, S.R.M.,
McConnaughey, T.A., Whelan, J.F., 1997. Calcification generates protons for nutrient 2011. Application of a yeast biosurfactant in the removal of heavy metals and
and bicarbonate uptake. Earth Sci. Rev. 42 (1e2), 95e117. hydrophobic contaminant in a soil used as slurry barrier. Appl. Environ. Soil Sci.
Meldrum, F.C., 2003. Calcium carbonate in biomineralisation and biomimetic 2011.
chemistry. Int. Mater. Rev. 48 (3), 187e224. Rusznya k, A., Akob, D.M., Nietzsche, S., Eusterhues, K., Totsche, K.U., Neu, T.R., et al.,
Millo, C., Dupraz, S., Ader, M., Guyot, F., Thaler, C., Foy, E., Me nez, B., 2012. Carbon 2012. Calcite biomineralization by bacterial isolates from the recently discov-
isotope fractionation during calcium carbonate precipitation induced by ure- ered pristine karstic Herrenberg cave. Appl. Environ. Microbiol. 78 (4),
olytic bacteria. Geochem. Cosmochim. Acta 98, 107e124. 1157e1167.
Ministerio de Obras Publicas, 2018. Direccio n de Vialidad. Gobierno de Chile. Seifan, M., Berenjian, A., 2019. Microbially induced calcium carbonate precipitation:
Retrieved from. http://www.vialidad.cl/Paginas/default.aspx. a widespread phenomenon in the biological world. Appl. Microbiol. Biotechnol.
Minto, J.M., Hingerl, F.F., Benson, S.M., Lunn, R.J., 2017. X-ray CT and multiphase flow 103 (12), 4693e4708.
characterization of a ‘bio-grouted’sandstone core: the effect of dissolution on Shahrokhi-Shahraki, R., Zomorodian, S.M.A., Niazi, A., O’Kelly, B.C., 2015. Improving
seal longevity. Int. J. Greenh. Gas Control 64, 152e162. sand with microbial-induced carbonate precipitation. Proc. Inst. Civ. Eng.
Mitchell, A.C., Dideriksen, K., Spangler, L.H., Cunningham, A.B., Gerlach, R., 2010. Ground Improv. 168 (3), 217e230.
Microbially enhanced carbon capture and storage by mineral-trapping and Sharma, A., Ramkrishnan, R., 2016. Study on effect of microbial induced calcite
 C.R.M. Portugal et al. / Journal of Cleaner Production 262 (2020) 121372 13

precipitates on strength of fine-grained soils. Perspect. Sci. 8, 198e202. 1837 (1), 12e19.
Sherard, J.L., Decker, R.S., 1977. Dispersive clays, related pipings and erosion in Wang, Y., Wang, H., Cheng, H., Chang, F., Wan, Y., She, X., 2020. Niche differentiation
geotechnical projects, 623. ASTM International. in the rhizosphere and endosphere fungal microbiome of wild Paris polyphylla
Soon, N.W., Lee, L.M., Khun, T.C., Ling, H.S., 2013. Improvements in engineering Sm. PeerJ 8, e8510.
properties of soils through microbial-induced calcite precipitation. KSCE J. Civ. Warren, L.A., Maurice, P.A., Parmar, N., Ferris, F.G., 2001. Microbially mediated cal-
Eng. 17 (4), 718e728. cium carbonate precipitation: implications for interpreting calcite precipitation
Stepkowska, E.T., Pe rez-Rodríguez, J.L., Sayagues, M.J., Martinez-Blanes, J.M., 2003. and for solid-phase capture of inorganic contaminants. Geomicrobiol. J. 18 (1),
Calcite, vaterite and aragonite forming on cement hydration from liquid and 93e115.
gaseous phase. J. Therm. Anal. Calorim. 73 (1), 247e269. Wei, S., Cui, H., Jiang, Z., Liu, H., He, H., Fang, N., 2015. Biomineralization processes of
Stocks-Fischer, S., Galinat, J.K., Bang, S.S., 1999. Microbiological precipitation of calcite induced by bacteria isolated from marine sediments. Braz. J. Microbiol.
CaCO3. Soil Biol. Biochem. 31 (11), 1563e1571. 46 (2), 455e464.
Teng, H., Kwigizile, V., James, D.E., Merle, R., 2007. Identifying influencing factors on Whiffin, V.S., 2004. Microbial CaCO3 Precipitation for the Production of Biocement.
paved roads silt loading. J. Air Waste Manag. Assoc. 57 (7), 778e784. Doctoral dissertation. Murdoch University.
Terzaghi, K., 1943. Theoretical Soil Mechanics. John Wiley & Sons, New York. Whiffin, V.S., Van Paassen, L.A., Harkes, M.P., 2007. Microbial carbonate precipita-
Terzaghi, K., Peck, R.B., Mesri, G., 1996. Soil Mechanics in Engineering. Practice. John tion as a soil improvement technique. Geomicrobiol. J. 24 (5), 417e423.
Wiley & Sons. Wichtmann, T., Triantafyllidis, T., 2009. Influence of the grain-size distribution
Tobler, D.J., Maclachlan, E., Phoenix, V.R., 2012. Microbially mediated plugging of curve of quartz sand on the small strain shear modulus G max. J. Geotech.
porous media and the impact of differing injection strategies. Ecol. Eng. 42, Geoenviron. Eng. 135 (10), 1404e1418.
270e278. Wong, L.S., 2015. Microbial cementation of ureolytic bacteria from the genus Ba-
Tsuneda, S., Jung, J., Hayashi, H., Aikawa, H., Hirata, A., Sasaki, H., 2003. Influence of cillus: a review of the bacterial application on cement-based materials for
extracellular polymers on electrokinetic properties of heterotrophic bacterial cleaner production. J. Clean. Prod. 93, 5e17.
cells examined by soft particle electrophoresis theory. Colloids Surf. B Bio- Wright, D.T., Oren, A., 2005. Nonphotosynthetic bacteria and the formation of
interfaces 29 (2e3), 181e188. carbonates and evaporites through time. Geomicrobiol. J. 22 (1e2), 27e53.
Umar, M., Kassim, K.A., Chiet, K.T.P., 2016. Biological process of soil improvement in Wu, J.R., Han, S.F., Zhu, Y.Y., Lu, M., Wang, G.P., Guo, W.L., 2006. Study on taxonomy
civil engineering: a review. J. Rock Mech. Geotech. Eng. 8 (5), 767e774. of endophytic fungi isolated from orchid mycorrhizae in Yunnan province.
Umesh, T., Dinesh, S., Sivapullaiah, P.V., 2011. Characterization of dispersive soils. J. Southw. For. Coll. 26 (3), 5e10.
Mater. Sci. Appl. 2 (6), 629e633. Yang, J., Gu, X.Q., 2013. Shear stiffness of granular material at small strains: does it
Vakili, A.H., Selamat, M.R., Moayedi, H., Amani, H., 2013. Stabilization of dispersive depend on grain size? Geotechnique 63 (2), 165e179.
soils by pozzolan. For. Eng. 2012: Gatew. Saf. Tomorrow 726e735. Yang, X., Xu, G., Chen, Y., Wang, F., Mao, H., Sui, W., et al., 2009. CaCO3 crystallization
Valencia, Y., 2009. Influe ^ncia da biomineralizaç~ ao nas propriedades físico-mec^ an- control by poly (ethylene oxide)epoly (propylene oxide)epoly (ethylene oxide)
icas de um perfil de solo tropical afetado por processos erosivos. Doctoral triblock copolymer and O-(hydroxy isopropyl) chitosan. J. Cryst. Growth 311
dissertation, PhD Thesis. Universidad de Brasilia, Brasilia, Brasil. (21), 4558e4569.
Valencia, Y., Camapum, J., Torres, F.A., 2014. Influence of biomineralization on the Yasuhara, H., Hayashi, K., Okamura, M., 2011. Evolution in mechanical and hydraulic
physico-mechanical profile of a tropical soil affected by erosive processes. Soil properties of calcite-cemented sand mediated by biocatalyst. Geo-Front. 2011:
Biol. Biochem. 74, 98e99. Adv. Geotech. Eng. 3984e3992.
Vallejo, L.E., Mawby, R., 2000. Porosity influence on the shear strength of granular Zhang, Q., Xu, Z., Shen, Z., Li, S., Wang, S., 2009. The Han River watershed man-
materialeclay mixtures. Eng. Geol. 58 (2), 125e136. agement initiative for the South-to-North water transfer project (Middle Route)
Van Paassen, L.A., 2009. Biogrout, Ground Improvement by Microbial Induced of China. Environ. Monit. Assess. 148 (1e4), 369e377.
Carbonate Precipitation [Ph. D. thesis]. Delft University of Technology, Delft, The Zhang, Q., Ren, L., Sheng, Y., Ji, Y., Fu, J., 2010. Control of morphologies and poly-
Netherlands. morphs of CaCO3 via multi-additives system. Mater. Chem. Phys. 122 (1),
Van Paassen, L.A., 2011. Bio-mediated ground improvement: from laboratory 156e163.
experiment to pilot applications. Geo-Front. 2011: Adv. Geotech. Eng. Zhu, T., Dittrich, M., 2016. Carbonate precipitation through microbial activities in
4099e4108. natural environment, and their potential in biotechnology: a review. Front.
Van Paassen, L.A., Daza, C.M., Staal, M., Sorokin, D.Y., van der Zon, W., van Bioeng. Biotechnol. 4, 4.
Loosdrecht, M.C., 2010. Potential soil reinforcement by biological denitrification. Ziegler, A.D., Sutherland, R.A., Giambelluca, T.W., 2000. Runoff generation and
Ecol. Eng. 36 (2), 168e175. sediment production on unpaved roads, footpaths and agricultural land sur-
Vizcarra, G.O.C., 2010. Aplicabilidade de cinzas de resíduo so lido urbano para base faces in northern Thailand. Earth Surf. Process. Landforms: J. Br. Geomorphol.
de pavimentos. Trabalho de conclusa ~o de curso de mestrado em engenharia em Res. Group 25 (5), 519e534.
civil. Universidade Cato  lica do Rio de JaneiroeRJ. Ziegler, A.D., Giambelluca, T.W., Sutherland, R.A., Vana, T.T., Nullet, M.A., 2001.
Wang, L., Roy, A., Seals, R.K., Metcalf, J.B., 2003. Stabilization of sulfate-containing Horton overland flow contribution to runoff on unpaved mountain roads: a case
soil by cementitious mixtures mechanical properties. Transport. Res. Rec. study in northern Thailand. Hydrol. Process. 15 (16), 3203e3208.