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Strength Variation of Microbial Concrete for Different Bacterium Percentages

Conference Paper · January 2020

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 Paper ID: 117

STRENGTH VARIATION OF MICROBIAL CONCRETE FOR

DIFFERENT BACTERIUM PERCENTAGES

Sudipto Nath Priyom1*, S. M. Sarwar Uddin2, Moinul Islam3 & Wahhida Shumi4
1
Former Lecturer, Department of Civil Engineering, Port City International University, Chittagong,
<sudiptonathpriyom@gmail.com>
2
UG Student, Department of Civil Engineering, Port City International University, Chittagong,
<smsarwar98@gmail.com>
3
Professor, Department of Civil Engineering, Chittagong University of Engineering & Technology,
Chittagong, < mislam@cuet.ac.bd>
4
Professor, Department of Microbiology, University of Chittagong, Chittagong,
<shumi@cu.ac.bd>
*Corresponding Author

ABSTRACT
Concrete is a strong, durable, relatively cheap construction material that is composed of cement, sand,
stone and aggregate. It has ultimate load bearing capacity under compression but it is weak in tension.
That’s why steel reinforced bars are embedded in concrete. However, cracks in concrete reduce the
durability of concrete structure. A noble technique has been developed in recent decades to repair
these cracks automatically by incorporating a biological agent such as Bacillus subtilis bacteria. This
study was aimed at finding the strength by applying bacterial culture integrated into concrete matrix to
act as a self-healing agent to seal cracks. 150 mm cubical concrete specimens were cast and cured for
3, 7, 14, and 28 days in plain water with and without using bacterial water. An optical density of
0.3±0.1 was used in this study. To find out the optimum bacterium concentration, 4 different ratios of
microbial culture to plain water (0:100, 25:75, 50:50, and 75:25) were taken and directly added to
concrete. Later on the specimens were subjected to compressive strength test periodically. From the
investigation, it has been revealed that compressive strength of concrete cube increases as the
bacterium concentration increases and the ratio 75:25 shows better performance against strength
deterioration under all curing conditions and curing ages.

Keywords: Calcite Precipitation, Compressive Strength, Microbial Concrete, OPC

INTRODUCTION
Concrete has become the most usable building material for infrastructure development in every
country. But it is also a very brittle material with low tolerance for strain. That well being said, it is
expected to crack with time. Some reasons behind these cracks are shrinkage, freeze-thaw
environment and mechanical compressive and tensile forces. However, these tiny micro-cracks on the
surface of concrete structure can make the whole structure vulnerable when water seeps into concrete,
corrodes the steel reinforcement and ultimate reduces the life time of concrete structure.

Although many physical and chemical treatments have been applied to decrease the susceptibility to
damage, these treatments are not fully useful because of their non-reversible action and limited long
term performances. Recently, a unique technique has been introduced, that is by incorporating
biological agent in concrete. According to the concept microbiology, special genuses of bacteria can
secret calcium carbonate as an extracellular product when proper nutrient media and environment are
available. This microbiologically induced calcite precipitation (MICP) can be used to repair the cracks
in cracks in concrete. This MICP technique will produce CaCO3 to heal cracks that appear on the
surface of concrete structure.

1
The aim of the experimental investigation is to evaluate the mechanical strength of concrete. OD600
0.3±0.1 of microbial culture was used in this study. No admixture was used. Mix design was based on
material properties and was derived from software analysis.

THE BIOLOGICAL SELF-HEALING PROCESS

Different mechanisms behind this bio-calcification process have been proposed but they have been a
matter of controversy over the years. According to some researchers, bacterial surfaces play an
important role in calcite precipitation (Fortin et al, 1997). The surface charge of micro-organisms is
negative which attracts Ca2+ ions from the environment and by series of complex bio-chemical
reactions, the precipitation deposits on cell surface.

Ca2++ Cell = Cell-Ca2+

Cell-Ca2++ H2CO3 = Cell-CaCO3

Some researchers believe that this calcite precipitation is an unwanted and accidental by-product of
metabolism (Knorre & Krumbein, 2000) while other believes that this precipitation is a specific
process with ecological benefits (Ehrlich 1996, Mc Connaughey & Whelan, 1997). Hence, this
process of calcite precipitation remained a matter of debate over the years. However, this calcite
precipitation can expand further as the bacteria produce calcite lattice food. The crystals expand until
the entire gap is filled. This inherent and biochemical process helps to improve performance of
concrete.

There are various types of bacteria which are used for making microbial concrete and help to improve
the concrete strength and durability. Following are the some of the bacteria used in the concrete.

 Bacillus pasteurii  Bacillus balodurans

 Bacillus sphaericus  Escherichia coli
 Bacillus subtilis  Bacillus cohnii

EXPERIMENTAL PROGRAM
The experimental work was carried out to study the different aspects of strength development of
microbial concrete in plain water over a period of 28 days. The variable parameters studied and the
materials involved were as follows:

MATERIALS
Culture and Growth of Bacillus subtilis
Bacillus subtilis strain 121 had been used in this study. Possible metabolism process regarding this
bacterial group is oxidative deamination of amino acids (Seshagiri, 2012). It was collected from
Microbiology Department, University of Chittagong. Media used was Nutrient Broth for B. subtilis
growth. Required amount of peptone, beef extract and NaCl were taken in a 3000 ml conical flask
first. Then required amount water was added at room temperature. Finally the solution was stirred
slowly to prepare nutrient broth media. It is necessary to make the media germ free. Sterilization
autoclaves are widely used in microbiology. Nutrient Broth media was sterilized about 2 hours. Then
the media became totally germ free. The next step was the addition of Bacillus subtilis spores. Spores
were introduced here by a needle in prepared media. Then the media with spores was taken into
refrigerator. Appropriate temperature had been maintained. This was the place where bacteria are kept
to germinate in a binary fission. Concentration of bacteria is the factors of growth phase of bacteria
and germination time. Four different concentrations of bacteria had been used in this study.

Cement
Ordinary Portland Cement (OPC) was used in casting the specimen. The minimum compression
strength that the cement will attain after 28 days of setting was 31.4 MPa and the specific gravity was
3.15.

2
Aggregate
Locally available natural sand passing through 4.75 mm sieve and retained on 0.075 mm sieve was
used as fine aggregate. Crushed stones of maximum nominal size of 12.5 mm size were used as coarse
aggregate. The specific gravity, fineness modulus, water absorption, bulk density and moisture
content of the aggregates were tested.
Water
Portable water having pH value 7 at zero turbidity confirming to the requirements of IS456-2000 was
taken.

VARIABLES
Concrete Quality
Three different grades of microbial concrete having OD600 0.3±0.1 were used. OPC concrete was cast
for comparing its properties with that of microbial concrete.
Curing Period
Specimens were tested periodically after the specified curing periods of 7, 14 and 28 days in plain
water.
Concrete Specimen
150 mm x 150 mm x 150 mm cube specimens were prepared following ASTM standard procedure.
Curing Environment
A total of 120 concrete cubes were cast in the laboratory. After casting, the specimens were kept at
27°C temperature and 90% relative humidity for 24 hours. After demoulding, all the specimens were
cured in plain water for 28 days at room temperature.
Microbial Ratios
4 different ratios of microbial culture to plain water (0:100, 25:75, 50:50 and 75:25) were chosen and
directly added to concrete.

CONCRETE MIX DESIGN

Concrete was designed on the basis of material properties. Trial mix designs were made to get rough
ideas of total aggregates and cements and thereby they were collected. Finally mix design is made by
software analysis (Conmixer V 1.0).

For a mix design of 20 MPa concrete the ratio of cement, fine aggregate, coarse aggregate was
derived as 1.0: 2.42: 2.8 with water cement ratio of 0.592 by mass. For 30 MPa concrete the ratio was
1.0: 1.57: 2.11 with water cement ratio of 0.445. And for 40 MPa concrete the ratio was 1.0: 1.2: 1.79
with water cement ratio of 0.38 by mass. 4 different ratios of microbial culture to plain water were
studied by mass in this experiment.

RESULT & DISCUSSION

Figures 1 to 4 represent the experimental outcome of compressive strength test of microbial concrete
for different bacterium percentages and curing periods. These figures clearly demonstrate that
concrete specimens with higher bacterium percentages gives lower strength deterioration for all
curing periods.

It has been revealed that compressive strength of microbial concrete having ratio 75:25 shows better
result than other microbial ratios. It is probably due to the higher concentration of calcite precipitation
in the concrete matrix. Table 1 gives the complete outcome of the experiment.

3
 45 50
20 MPa 30 MPa 40 MPa 45 20 MPa 30 MPa 40 MPa
40
Compressive Strength (MPa)

40

Compressive Strength (MPa)

35
35
30
30
25
25
20
20
15
15
10
10
5 5
0 0
0 10 20 30 0 10 20 30
Curing Periods (Days) Curing Periods (Days)

[Fig- 1: Microbial Culture : Plain Water= 0 : 100] [Fig- 2: Microbial Culture : Plain Water= 25 : 75]

60 60
20 MPa 30 MPa 40 MPa 20 MPa 30 MPa 40 MPa
50 50
Compressive Strength (MPa)

Compressive Strength (MPa)

40 40

30 30

20 20

10 10

0 0
0 10 20 30 0 10 20 30
Curing Periods (Days) Curing Periods (Days)

[Fig- 3: Microbial Culture : Plain Water= 50 : 50] [Fig- 4: Microbial Culture : Plain Water= 75 : 25]

Strength increases up to 3.14% for 20 MPa concrete, 11.76% for 30 MPa concrete and 10.32% for 40
MPa concrete using the ratio of microbial water to plain water 25:75 for 28 days curing period. For
the ratio 50:50, it increases up to 18.32% for 20 MPa concrete, 17.64% for 30 MPa concrete and
23.17% for 40 MPa concrete. In case of the ratio 75: 25, strength increases up to 26.7% for 20 MPa
concrete, 30.1% for 30 MPa concrete and 27.46% for 40 MPa concrete.

4
 Table- 1: Strength Behavior Observation
Increased in compressive strength compared to
Ratio of microbial culture the ratio 0:100
to plain water 20 MPa 30 MPa 40 MPa
25: 75 3.14% 11.76% 10.32%
50: 50 18.32% 17.64% 23.17%
75: 25 26.70% 30.10% 27.46%

CONCLUSION
In this experiment, self-healing agent has been incorporated to enhance the strength of concrete and
significant increase in strength development has been observed. This MICP technique can be proved
to better than other conventional technologies due to its eco-friendly nature and self-healing activities.
This study may provide sufficient information regarding the optimum use of bacterium percentages.
1. The significance of this research is that calcite precipitation by Bacillus subtilis is effective in
increase in strength.
2. Significant amount of money can be saved as labor intense check in repairing cracks
minimizes.
3. Mix proportion of microbial water with plain water has a great impact on strength
development of microbial concrete. Among the microbial concretes, concrete with the ratio
75:25 is found to be most effective in increasing compressive strength.
4. It has been observed from table 1 that strength increases in a slow manner in case of lower
bacterium concentration. But as soon as the concentration increases, strength also increases.

REFERENCES
Anagnostopoulos, C., and Spizizen, J.,(1961), Requirements for Transformation in Bacillus Subtilis. J.
Bacteriol, 81:741-746
Anderson, S., Appanna, V. D., Huang, J., and Viswanatha, T.,(1992), A Novel Role for Calcite in
Calcium Homeostasis, FEBS Lett, 308:94-96
Barabesi, C., Tamburini, E., Mastromei, G., and Perito, B.,(2003), Mechanisms of Microbial Calcium
Carbonate Precipitation, p. 472-485, In Koestler, R. J., Koestler, V.
Bazylinski, D. A., and Moskowitz, B. M.,(1997), Microbial Biomineralization of Magnetic Iron
Minerals: Microbiology, Magnetism and Environmental Significance, Rev. Mineral, Geochem,
35:181-223
Bäuerlein, E.,(2003), Biomineralization of Unicellular Organisms: Unusual Membrane Biochemistry
for the Production of Inorganic Nano- and Microstructures,Angew. Chem. Int. Ed.42:614-641
Beveridge, T. J.,(1989), Role of Cellular Design in Bacterial Metal Accumulation and Mineralization,
Annu. Rev. Microbiol. 43:147-171
Castanier, S., Le Métayer-Levrel, G., Orial, G.,Loubière, J. F., and Perthuisot, J. P.,(2000), Bacterial
Carbonatogenesis and Applications to Preservation and Restoration of Historic Property, p. 203-
218, In Ciferri, O., Tiano, P., and Mastromei (ed.), G.,Of Microbes and Art: The Role of Microbial
Communities in the Degradation and Protection of Cultural Heritage, Plenum Publishers, New York,
NY
Douglas, S., and Beveridge, T. J.,(1998), Mineral Formation by Bacteria in Natural Microbial
Communities, FEMS Microbiol. Ecol. 26:79-88
Ehrlich, H. L.,(1996),Geomicrobiology. Marcel Dekker, New York
Ehrlich, H. L.,(1999), Microbes as Geologic Agents: Their Role in Mineral Formation, Geomicrobiol.
J. 16:135-153
Fortin, D., Ferris, F. G., and Beveridge, T. J.,(1997), Surface-Mediated Mineral Development by
Bacteria, Rev. Mineral. Geochem. 35:161-180
Gavimath, C. C., Mali, B. M., Hooli, V. R., Mallpur, J. D., Patil, A. B., Gaddi, D. P., Ternikar, C.R.,
Ravishankera, B.E., Potential Application of Bacteria to Improve The Strength of Cement Concrete,
International Journal of Advanced Biotechnology and Research, ISSN 0976-2612, Vol 3, Issue 1,
2012, pp 541-544

5
 Geymayr, August, G. W.,(1980). Repair of Concrete in Tropical Marine Environment, Performance
of Concrete in Marine Environment, ACI Publication, SP-65, pp. 527-556
Manikandan, A.T., Padmavathi, A., An Experimental Investigation on Improvement of Concrete
Serviceability by using Bacterial Mineral Precipitation, Volume II, Issue III, March 2015, IJRSI,
ISSN 2321 – 2705
Meldrum F. C., (2003), Calcium Carbonate in Biomineralisation- Biomimetic Chemistry, 48, 187-224
Mohini P. Samudre, M. P., Mangulkar, M. N., Saptarshi, S. D., A Review of Emerging Way to
Enhance the Durability and Strength of Concrete Structures: Microbial Concrete. Vol. 3, Issue 2,
February 2014
Neville, A., Autogenous healing – A concrete miracle?, Concrete International, November 2002
Ramos-Cormenzana, A.,(1975), Formation of Calcite Crystals by Bacteria of the Genus Bacillus,
Microbios 13:61-70
Ravindranatha, Kannan, N., Likhit M. L., Self-healing Material Bacterial Concrete, IJRET:
International Journal of Research in Engineering and Technology, eISSN: 2319-1163 | pISSN: 2321-
7308
Reddy, S., Rao, M. V. S., Satya, A. , Azmatunnisa, M., A Biological Approach to Enhance Strength
And Durability in Concrete Structure, International Journal of Advances in Engineering &
Technology, Sept 2012 ©IJAET, ISSN: 2231-1963
Santhosh K. R., Ramakrishnan V., Duke E. F., and Bang S.S., SEM Investigation of Microbial
Calcite Precipitation in Cement, Proceedings of the 22nd International Conference on Cement
Microscopy, pp. 293-305, Montreal, Canada, 2000
Schlangen, E., Sangadji, S., Addressing Infrastructure Durability and Sustainability by Self Healing
Mechanisms- Recent Advances in Self Healing Concrete and Asphalt, The 2nd International
Conference on Rehabilitation and Maintenance in Civil Engineering, Procedia Engineering 54
(2013) 39 – 57
Von Knorre, H., Krumbein, W. E.,(2000), Bacterial Calcification, p. 25-31,InRiding, R. E., and
Awramik (ed.), S. M., Microbial Sediments, Springer-Verlag, Berlin, Germany

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