Construction and Building Materials 121 (2016) 659–663

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Construction and Building Materials

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

Influences of bacteria-based self-healing agents on cementitious

materials hydration kinetics and compressive strength
Mian Luo, Chunxiang Qian ⇑
College of Materials Science and Engineering, Southeast University, Nanjing 211189, China
Research Institute of Green Construction Materials, Southeast University, Nanjing 211189, China

h i g h l i g h t s

 The hydration kinetics of bacteria-based cement paste was studied.

 The compressive strength of bacteria-based cement mortar was tested.
 The compatibility of bacteria-based cementitious materials was evaluated.

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

Article history: Bacterially induced calcium carbonate precipitation has been proposed as an alternative and environ-
Received 29 October 2015 mental technique to develop self-healing cementitious materials system in recent years. This study inves-
Received in revised form 2 June 2016 tigated the influences of bacteria-based self-healing agents on the rheology, hydration kinetics and
Accepted 14 June 2016
compressive strength of cementitious materials to further verify the feasibility of bacteria-based self-
healing agents for crack repairing. The results showed that the rheology of cement mortar was signifi-
cantly improved by the addition of bacteria-based self-healing agents. Incorporation of bacteria-based
Keywords:
self-healing agents in cement greatly influenced the hydration kinetics. The self-healing agent RB could
Bacteria
Self-healing agent
delay the hydration of cement resulting in final setting time increase, however the self-healing agents JB
Rheology and NB accelerated the hydration resulting in initial and final setting time decrease. In addition, compres-
Hydration heat sive strength test results showed that incorporation of RB in cement mortar resulted in early age com-
Setting time pressive strength decrease, but the 28 d compressive strength increased compared to control.
Compressive strength Incorporation of JB in cement mortar enhanced compressive strength, however incorporation of NB in
cement mortar could result in unwanted compressive strength loss. Furthermore, MIP test results indi-
cated that the pore size distributions were different between cement paste samples with and without
bacteria-based self-healing agents. The addition of bacteria-based self-healing agents increased the
porosity between 100 and 1000 nm. The porosity between 100 and 1000 nm of the sample with NB
increased the most compared to the samples with RB and JB. The incorporation of JB increased the poros-
ity between 100 and 1000 nm, but at the same time the small pores with a diameter under 10 nm
increased. These results could be used to explain the compressive strength development.
Ó 2016 Elsevier Ltd. All rights reserved.

1. Introduction a specific substrate should be added into cementitious materials

Bacterially induced calcium carbonate precipitation has been
proposed as an alternative and environmental technique to
develop a self-healing cementitious materials system in recent
years [1–7]. In this system, the bacteria-based self-healing agent
which consists of the alkali resistance spore-forming bacteria and
⇑ Corresponding author at: College of Materials Science and Engineering,
Southeast University, Nanjing 211189, China.
E-mail addresses: cxqian@seu.edu.cn, cquluomian@163.com (C. Qian).
in the mixing process. The bacteria can form dormant spores in
extreme environments of cementitious materials. When the con-
crete cracks, the dormant spores could be activated by the water
and oxygen entering through the cracks. Then, the substrate in
the crack migrating from cementitious materials matrix could be
metabolized by the activated bacteria to produce calcium carbon-
ate precipitation after a series of biochemical reactions and achieve
the purpose of crack self-healing. A large number of studies on this
topic have been carried out, mainly focusing on the effect of crack
self-healing [8,9], permeability test [10,11], the mechanism of

http://dx.doi.org/10.1016/j.conbuildmat.2016.06.075
0950-0618/Ó 2016 Elsevier Ltd. All rights reserved.
660 M. Luo, C. Qian / Construction and Building Materials 121 (2016) 659–663

mineralization [12], optimization of nutrients [5,13], bacteria spe- Table 1

cies and carrier selection [14–16]. The results showed that Mixing proportion of cement mortar.

bacteria-based self-healing agents could improve the ability of Code Cement Sand Water Self-healing Self-healing agent
cementitious materials crack self-repairing and reduce the possi- (g) (g) (g) agent types contents (wt%)
bility of erosion media through the cracks into the matrix resulting C 450 1350 225 / 0
in improvement of the material durability. However, the prerequi- RB1 450 1350 225 RB 1
site for achieving this goal is the need to introduce bacteria-based RB2 450 1350 225 RB 2
RB3 450 1350 225 RB 3
self-healing agents as a new component into cementitious materi- JB1 450 1350 225 JB 1
als. Cementitious materials is a kind of complex multi-component JB2 450 1350 225 JB 2
composite material and anyone new component introducing could JB3 450 1350 225 JB 3
have adversely affected its performance. So, it is very important to NB1 450 1350 225 NB 1
NB2 450 1350 225 NB 2
investigate the compatibility between bacteria-based self-healing
NB3 450 1350 225 NB 3
agents and cementitious materials. Jonkers et al. [17] investigated
the effect of bacteria and organics additions on paste strength.
Incorporation of a high number of bacterial spores in the paste
The initial and final setting times of the cement paste with and without self-
resulted in about 10% decrease in compressive strength compared healing agents RB, JB and NB in various contents from 1% to 3% were tested, respec-
to controls. Additions of different organic compounds (0.5% of tively. In order to reveal the influence mechanism of setting time, the effects of each
cement weight) also affected splitting-tensile strength of the paste. component of self-healing agents on setting time were researched.
The study results of Zeynep Basaran Bundur et al. [18] indicated
that hydration kinetics and compressive strength behavior of mor- 2.4. Hydration heat
tar were highly influenced when Sporosarcina pasteurii cells were
A constant temperature gauge TAM Air (Swedish Leite La Ltd) was used to mea-
vegetatively inoculated.
sure the rate of heat evolution and the total heat evolution during 72 h of cement
In our previous studies, three types of bacteria-based self- hydration, the experimental temperature is kept at 20 °C. The cement paste mixing
healing agents were developed. The crack self-healing capacity of was performed by water to cement ratio of 0.5. The hydration heat evolutions of the
bacteria-based concrete in different conditions has been widely cement paste with and without self-healing agents RB, JB, NB, R, J, N and B in con-
researched and the results showed that the bacteria-based self- tents of 2% were investigated, respectively.

healing agents could be used to achieve the goal of concrete crack

self-healing [19]. This study investigated the influences of three
types of bacteria-based self-healing agents on the rheology, hydra-
tion kinetics and compressive strength of cementitious materials
to further verify the feasibility of bacteria-based self-healing
agents for crack repairing. Moreover, the influence mechanism of
bacteria-based self-healing agents on cementitious materials was
discussed in detail.
2.5. Mercury intrusion porosimetry (MIP)
In order to explain the influences of bacteria-based self-healing agents on the
properties of the cement mortar, the pore size distribution test of cement paste
(water to cement ratio of 0.4) with different self-healing agents (RB, JB and NB)
was performed using MIP. MIP tests were made with a Micrometrics AutoPore IV
9500 (American Michael Instruments Corp., USA), which has a pore measuring
range between 0.0036 and 400 lm diameters. After curing for 28 days, the samples
were broken and dried for 3 days in the oven.

2. Materials and methods

3. Results and discussion
2.1. Bacteria-based self-healing agents preparation
3.1. Rheology of cement mortar
Three types of bacteria-based self-healing agents (RB, JB and NB) were used in
this study. RB agent was consisted of calcium lactate (R) and bacteria spores powder
(B), JB agent was consisted of calcium formate (J) and bacteria spores powder (B)
The fluidity test results of cement mortar with bacteria-based
and NB agent was consisted of calcium nitrate (N) and bacteria spores powder self-healing agents RB, JB and NB in different dosages are presented
(B). Spore-forming alkali-resistant bacteria were used for this study. The bacteria in Fig. 1. It can be seen that cement mortar fluidity had been
were cultured in liquid medium containing 5.0 g peptone and 3.0 g yeast extract improved after the incorporation of bacteria-based self-healing
per liter of distilled water (pH = 7.0), which was autoclaved at 121 °C for 25 min.
agents. However, various improvement trends were found in
After inoculation on laminar flow, the medium was incubated at 30 °C on a shaker
at 170 rpm for 72 h. Bacterial cells were harvested by centrifuging the 48 h old
grown culture and were re-suspended in distilled water. Then the bacteria cells sus-
pension liquid was kept in 80 °C water baths for 20 min to get bacteria spores sus-
pension liquid. The concentration of bacterial spores in the suspension was 190 RB
108 cells/mL. The bacteria spore powder was obtained by drying of bacteria spores
JB
suspension liquid.
185 NB
2.2. Preparation of cement mortar specimens
180
Fluidity (mm)

Cement mortar specimens with dimensions of 40  40  160 mm were pre-

pared by mixing ordinary Portland cement, river sand, self-healing agent and tap
water. The mixing proportion is shown in Table 1. The self-healing agent to cement
175
ratio is 0%, 1%, 2% and 3%. The cement mortar fluidity was measured before casting
according to the Chinese standard (GB/T 2419-2005). After 24 h curing specimens
were unmolded and kept in standard curing room for further curing. In order to
170
determine the influences of bacteria-based self-healing agents on compressive
strength of cement mortar, sets of 3 replicate specimens were tested after1, 3, 7
and 28 days curing.
165

2.3. Setting time

0 1 2 3
The initial and final setting times were measured by the technique for deter- Self-healing agents contents (wt.%)
mining the time of setting of hydraulic cement using a Vicat needle according to
Chinese standard (GB/T 1346-2011). Fig. 1. Results of the fluidity test of cement mortar.
 M. Luo, C. Qian / Construction and Building Materials 121 (2016) 659–663 661

different bacteria-based self-healing agents with the increase of

content. The highest fluidity value was obtained in the cement mor-
tar with self-healing agent RB when the dosage of self-healing agent
was 1%, but the cement mortar with self-healing agent NB pre-
sented the highest fluidity value as the dosage up to 2% and 3%. This
may be attributable to the combined action of bacteria and sub-
strate in self-healing agents added to the cement mortar.

3.2. Setting time of cement paste

Figs. 2–4 show the initial and final setting time of the cement
paste with self-healing agents RB, JB and NB in different contents,
respectively. The results indicated that addition of self-healing
agent RB had an obvious setting retarding. With the increase of
RB content, the initial setting time gradually decreased, but the
final setting time significantly increased. The initial and final times
of control samples were 136 min and 216 min, respectively,
whereas for the cement paste with RB in contents of 3%, the initial
setting time was 68 min with final setting at 338 min. However, Fig. 3. Initial and final setting time of cement mortar with different JB contents.
the addition of JB and NB accelerated the setting and the initial
and final setting time gradually decreased with the increase of JB
and NB contents, respectively. In order to reveal the influence
mechanism of setting time, the effects of each component of self-
healing agents on setting time were researched, as shown in
Fig. 5. Through a detailed study of the individual component of
self-healing agents, the results showed setting retarding mainly
caused by the calcium lactate and the setting acceleration could
be attributed to calcium formate and calcium nitrate. In addition,
bacteria spore powder had only a slight setting retarding effect.

3.3. Hydration heat of cement paste

Heat generation date during early hydration of cement paste

with and without self-healing agents are shown in Figs. 6 and 7.
It can be seen that addition of self-healing agents influence the
hydration kinetics of cement pastes. The heat flow evolution dur-
ing early hydration of control has a peak at 10.5 h. However, the
heat flow evolution of cement paste with JB, NB, J and N has a peak
at around 6 h and the cement paste with RB and R at around 16.5 h,
Fig. 4. Initial and final setting time of cement mortar with different NB contents.
indicating that JB and NB could accelerate hydration and RB delay
hydration. Moreover, the results show that bacteria spore powder
has little influence on cement hydration and the hydration acceler-
ation mainly owe to the calcium formate and calcium nitrate, but
the hydration delay could be attributed to calcium lactate. The
results were consistent with the setting time test results above.

400 ,QLWLDOWLPH5%
)LQDOWLPH5%
350

300
Setting time (min)

250

200

150

100
Fig. 5. Initial and final setting time of cement mortar with each component of self-
50 healing agents.

0
0 1 2 3 From Fig. 7, it can be found that the total heat generated before
20 h of hydration for cement pastes with JB, NB, J and N was higher
Self-healing agent contents (wt.%)
than the control and the cement paste with RB and B was lower
Fig. 2. Initial and final setting time of cement mortar with different RB contents. than the control. However, the total heat generated during 72 h
662 M. Luo, C. Qian / Construction and Building Materials 121 (2016) 659–663

Fig. 6. Heat flow during the hydration of cement pastes with and without self-
healing agents. Fig. 8. Compressive strength development of cement mortar samples with RB.

Fig. 7. Total heat during the hydration of cement pastes with and without self- Fig. 9. Compressive strength development of cement mortar samples with JB.
healing agents.

of hydration for all cement pastes with self-healing agents was

lower than the control except the cement paste with B.

3.4. Compressive strength of cement mortar

Figs. 8–10 show that the results of 1 d, 3 d, 7 d and 28 d com-

pressive strength tests for the cement mortar samples with self-
healing agents RB, JB and NB, respectively. Incorporation of RB in
cement mortar resulted in early age compressive strength
decrease, but the 28 d compressive strength increased compared
to control. With the increase of RB contents from 1% to 3%, the
1 d compressive strength decreased from 13.32 N/mm2 to 5.37 N/
mm2 due to setting retarding action of RB, however the 28 d com-
pressive strength was improved. This needs to be stressed that the
decrease of early age compressive strength was acceptable when
the content of RB is less than 2%. From Fig. 9, it can be found that
incorporation of JB in cement mortar enhanced compressive
Fig. 10. Compressive strength development of cement mortar samples with NB.
strength. However, the enhancement effect of 28 d compressive
strength decreased with the increase of JB content. Incorporation
of NB in cement mortar could result in unwanted compressive NB with the content of 3% in cement mortar resulted in about
strength loss. The compressive strength was nearly the same with 36.3%, 26.8%, 25.1% and 17.4% decrease in compressive strength
the control when the content of NB is 1%, however incorporation of after 1, 3, 7 and 28 days curing, respectively.
 M. Luo, C. Qian / Construction and Building Materials 121 (2016) 659–663 663

loss. Furthermore, MIP test results indicated that the pore size dis-
tributions were different between cement paste samples with and
without bacteria-based self-healing agents. In control sample the
majority of the pores concentrate on diameters between 10 and
100 nm whereas the addition of bacteria-based self-healing agents
increased the porosity between 100 and 1000 nm. The porosity
between 100 and 1000 nm of sample with NB increased the most
compared to the samples with RB and JB. The incorporation of JB
increased the porosity between 100 and 1000 nm, but at the same
time the small pores with a diameter under 10 nm increased. These
results could be used to explain the compressive strength test
results.

Acknowledgments

The financial help of the National Nature Science Foundation of

China (Grant No. 51178104), 333 Project of Jiangsu Province and
Ph.D. Program’s Foundation of Ministry of Education of China
(Grant No. 20110092110033) is gratefully acknowledged.

Fig. 11. Pore size distribution curve of the cement pastes with and without
bacteria-based self-healing agents (curing for 28 days).
3.5. The pore size distribution of cement pastes
The pore size distribution curves of cement paste samples with
different bacteria-based self-healing agents (RB, JB and NB) were
shown in Fig. 11. It can be seen that the pore size distributions
were different between the samples with and without bacteria-
based self-healing agents. In control sample the majority of the
pores concentrate on diameters between 10 and 100 nm whereas
the addition of bacteria-based self-healing agents increased the
porosity between 100 and 1000 nm. The porosity between 100
and 1000 nm of sample with NB increased the most compared to
the samples with RB and JB. The incorporation of JB increased
the porosity between 100 and 1000 nm, but at the same time the
small pores with a diameter under 10 nm increased. The results
were consistent with the strength test results, which could well
explain why the addition of self-healing agent JB had improved
the 28 days compressive strength of cement mortar, however the
addition of self-healing agent NB resulted in unwanted compres-
sive strength loss.
4. Conclusion
The study investigated the influences of bacteria-based self-
healing agents on the rheology, hydration kinetics and compres-
sive strength of cementitious materials to further verify the feasi-
bility of bacteria-based self-healing agents for crack repairing.
Based on the present experimental investigation, the following
conclusions are drawn. The rheology of cement mortar was signif-
icantly improved by addition of the bacteria-based self-healing
agents. Incorporation of bacteria-based self-healing agents in
cement greatly influenced the hydration kinetics. The self-healing
agent RB could delay the hydration of cement resulting in final set-
ting time increase, however the self-healing agents JB and NB
accelerated the hydration resulting in initial and final setting time
decrease. In addition, compressive strength test results showed
that incorporation of RB in cement mortar resulted in early age
compressive strength decrease, but the 28 d compressive strength
increased compared to control. Incorporation of JB in cement mor-
tar enhanced compressive strength, however incorporation of NB
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in cement mortar could result in unwanted compressive strength