Sei sulla pagina 1di 12

123

Journal of The Institution of Engineers (India): Series A

Civil, Architectural, Environmental and Agricultural Engineering

ISSN 2250-2149

J. Inst. Eng. India Ser. A DOI 10.1007/s40030-017-0227-x

Performance of Microbial Concrete Developed Using Bacillus Subtilus JC3

M. V. Seshagiri Rao, V. Srinivasa Reddy & Ch. Sasikala

123 Journal of The Institution of Engineers (India): Series A Civil, Architectural, Environmental and Agricultural Engineering

the published article

123

Your article is protected by copyright and all rights are held exclusively by The Institution

of Engineers (India). This e-offprint is for personal use only and shall not be self- archived in electronic repositories. If you wish to self-archive your article, please use the accepted manuscript version for posting on your own website. You may further deposit the accepted manuscript version in any repository, provided it is only made publicly available 12 months after official publication or later and provided acknowledgement is given to the original source of publication

and a link is inserted to

on Springer's website. The link must be accompanied by the following text: "The final publication is available at link.springer.com”.

Author's personal copy

J. Inst. Eng. India Ser. A DOI 10.1007/s40030-017-0227-x

Author's personal copy J. Inst. Eng. India Ser. A DOI 10.1007/s40030-017-0227-x ORIGINAL CONTRIBUTION Performance of Microbial

ORIGINAL CONTRIBUTION

 

Performance of Microbial Concrete Developed Using Bacillus Subtilus JC3

M. V. Seshagiri Rao 1 V. Srinivasa Reddy 2 Ch. Sasikala 3

Author's personal copy J. Inst. Eng. India Ser. A DOI 10.1007/s40030-017-0227-x ORIGINAL CONTRIBUTION Performance of Microbial

Received: 23 May 2013 / Accepted: 9 October 2017 The Institution of Engineers (India) 2017

Abstract Concrete is vulnerable to deterioration, corro- sion, and cracks, and the consequent damage and loss of strength requires immensely expensive remediation and repair. So need for special concrete that they would respond to crack formation with an autonomous self-heal- ing action lead to research and development of microbial concrete. The microbial concrete works on the principle of calcite mineral precipitation by a specific group of alkali- resistant spore-forming bacteria related to the genus Bacillus called Bacillus subtilis JC3, this phenomenon is called biomineralization or Microbiologically Induced Calcite Crystal Precipitation. Bacillus subtilis JC3, a common soil bacterium, has inherent ability to precipitate calcite crystals continuously which enhances the strength and durability performance of concrete enormously. This microbial concrete can be called as a ‘‘Self healing Bac- terial Concrete’’ because it can remediate its cracks by itself without any human intervention and would make the concrete more durable and sustainable. This paper discuss the incorporation of microorganism Bacillus subtilis JC3 (developed at JNTU, India) into concrete and presents the results of experimental investigations carried out to study the improved durability and sustainability characteristics of microbial concrete.

& M. V. Seshagiri Rao rao_vs_meduri@yahoo.com

  • 1 Department of Civil Engineering, Jawaharlal Nehru Technological University Hyderabad College of Engineering, Hyderabad, India

  • 2 Department of Civil Engineering, Gokaraju Rangaraju Institute of Engineering and Technology, Hyderabad, India

  • 3 Centre for Environment, Jawaharlal Nehru Technological University Hyderabad , Hyderabad, India

Keywords Bacillus subtilis JC3 Bio-mineralization Self healing Bacterial concrete RCPT

Introduction

Crack repair is of crucial importance to concrete since cracks are the main path for the penetration of corrosive substances which greatly decrease the service life of con- crete structures. Nowadays, a promising method to repair concrete cracks is being investigated. The idea is that repair work should be done by the concrete itself when cracks appear, through a self-healing process by a certain kinds of natural healing agents released within the concrete on occurrence of cracks [1, 2]. A biological approach of self healing in concrete is one such crack remediation methods in which bacteria Bacillus subtilis JC3 can produce or induce bio-minerals during their growth and metabolism to fill the cracks and pores in the concrete. Concrete is an alkaline material whose pH is as high as 13. This alkalinity imparts concrete the protection against corrosion. Bacillus subtilis JC3 is an alkaliphile which can survive in high alkaline concrete and just needs nutrients for microbial activities to form inorganic solids. Bacteria-based self- healing concrete is produced by incorporating spores of bacteria of a special kind (Bacillus subtilis JC3), in the concrete matrix at the stage of preparation of the concrete by mixing the spore suspension in concrete mixing water. When crack is formed water enters the crack subsequently the homogeneously distributed bacterial spores in hardened concrete matrix gets activated and germinate to become metabolically active vegetative cells that are able to con- vert the organic nutrient compounds into insoluble inor- ganic calcium carbonate based minerals [3]. Copious

Author's personal copy J. Inst. Eng. India Ser. A DOI 10.1007/s40030-017-0227-x ORIGINAL CONTRIBUTION Performance of Microbial

123

Author's personal copy

J. Inst. Eng. India Ser. A

production of bacteria-mediated minerals results in sealing off the crack; hence the permeability and porosity of con- crete are reduced significantly [4]. The precipitated calcite has a coarse crystalline structure and is highly insoluble in water. It resists the penetration of harmful agents (chlo- rides, sulphates, carbon dioxide) into the concrete thereby decreasing the deleterious effects they cause [5, 6].

Bacterial Strain—Bacillus Subtilis JC3

Strain Bacillus subtilis JC3, selected for the present study, was distinguished as aerobic alkaliphilic spore-forming soil bacteria. The medium used to grow Bacillus subtilis JC3 was based on peptone, NaCl, yeast extract. The pure cul- ture was isolated from the soil sample of JNTU, India.

Chemistry of MICCP by Bacillus Subtilis JC3

In nature, microorganisms can induce calcite mineral pre- cipitation through nitrogen cycle either by ammonification of amino acids/nitrate reduction/hydrolysis of urea. Bacil- lus subtilis JC3 is able to precipitate calcium carbonate (CaCO 3 ) in its micro-environment by the ammonification of amino acids into ammonium (NH 4 ? ) and carbonate (CO 3 2 - ) ions. The precipitated bio-CaCO 3 has a great potential ability to heal concrete cracks because it is nat- ural, environmentally friendly and compatible with the concrete matrix. Bio-mineralization by Ammonification (Ammo acid degradation) comprises of series of complex biochemical reactions. Amino acids released during pro- teolysis (the process of enzymatic breakdown of proteins by the microorganisms with the help of proteolysis enzymes) undergo deamination in which nitrogen con- taining amino (–NH 2 ) group is removed. Thus, process of deamination which leads to the production of ammonia is termed as ‘‘ammonification’’. The process of ammonifica- tion is mediated by Bacillus subtilis JC3. Ammonification usually occurs under aerobic conditions (known as oxida- tive deamination) with the liberation of ammonia (NH 3 ) or ammonium ions (NH 4 ) when dissolved in water. The pro- cess of ammonification is represented as follows,

obtained precipitation without the application of calcino- genic bacteria. In addition to the bio-deposition treatment, the use of bacterially induced carbonates as a binder, i.e. bio-cementation, has been addressed. Until now, work on bio-deposition was mainly concentrated in Europe, while much of the work on remediation of cracks in concrete has been done in the USA. In China, researchers are investi- gating the use of microbially induced carbonate precipita- tion for the restoration of ancient masonry buildings [7] and the protection of concrete surfaces [8]. Furthermore, the method of producing CaCO 3 by bacterial biomineral- ization has been patented in China by Qian et al. [9]. The latter was possible, as the patent by Adolphe et al. [10] was limited to European countries. In Brazil, Shirakawa et al. [11] are working on bio-deposition on fiber cement roof tiles. In India, researchers have applied for a patent in which the use of Shewanella for the improvement of the strength of concrete was described [12]. Research and development of bacterial concrete was happening for the past 10 plus years in various universities outside India. In India very limited research was done in the area of biore- mediation of concrete particularly. The pioneering research was conducted on self-healing phenomena in bio-concrete by Bang et al. [13] at South Dakota School of Mines and Technology, USA. Willem De Muynck, Nele De Belie, Willy Verstraete, Kim Van Tittelboom et al. at Ghent University, Belgium has done extensive research on microbiologically induced calcite precipitation in con- struction materials. Henk M. Jonkers et al. at Delft University of Technology, Netherlands has studied exten- sively on the self-healing capability of bacteria induced cementitious materials. In India, P. Ghosh, S. Mandal, B. D. Chattopadhyay et al. of Jadavpur University, Kolkata, India; V Achal, Abhijeet Mukerjee, Rafat Siddique et al. of Thapar University, Patiala, India and M V Seshagiri Rao, V S Reddy et al. of Jawaharlal Nehru Technological University (JNTU), Hyderabad, India have done extensive research on the development of high performance self-re- mediating bacterial concrete. Researchers around the world attempted to study about the application of bio-mineral- ization in civil engineering specifically about the use of

ð

CH 3 CH NH

  • 2 ÞCOOH ðPeptoneÞþ 1 = 2 O 2 ! C 2 H 2 þ H 2 CO 3 þ NH 3 ðammoniaÞ

particular bacteria in cementitious materials to self-heal

H 2 CO 3 ! H þ þ HCO

  • 3 and seal cracks without human intervention. Available

þ

NH 3 þ H 2 O ! NH þ OH

4

Literature Review

Different research groups have searched for alternative approaches to obtain a protective layer of calcium car- bonate on the surface of building materials (bio-deposi- tion). Some authors suggested the use of alternative microorganisms or metabolic pathways, while others

literature has not reported any such suitable self-healing system which has features such as long-term compatibility, eco-friendliness, good bonding with surrounding cement matrix, less human intervention, inexpensive and organic in nature. Though it is reported that the use of specific alkaliphilic mineral forming bacteria enhances the prop- erties of cement mortar but there exists little understanding of the effect of bacteria on the mechanical and durability properties of concrete. The significance of this research was to quantitatively understand and assess the role of

123

Author's personal copy

J. Inst. Eng. India Ser. A

Table 1 Mix proportions

Ordinary grade concrete (M20) 1: 2.27: 3.45: 0.54 Standard grade concrete (M40) 1: 1.73: 2.60: 0.42

High strength grade(M60)

High strength grade(M80)

1:1.25:2.41:0.26

(Micro Silica—6% bwc*) (Superplasticizer(SP)—1% bwc*)

1:1.06:1.96:0.23

(Micro Silica—10% bwc*) (Superplasticizer(SP)—1.2% bwc*)

* bwc-by weight of cement

microbiologically induced calcium carbonate precipitation in enhancing the mechanical and durability characteristics of concrete incorporated with Bacillus subtilis JC3.

Author's personal copy J. Inst. Eng. India Ser. A Table 1 Mix proportions Ordinary grade concrete

Fig. 1 Bacillus subtilis JC3

Experimental Investigations

Mix Proportions

The grades of concrete used in the present investigation are ordinary grade (M20), standard grade (M40) and high strength grade concretes (M60 and M80). The mix pro- portions of M20 and M40 grade concrete are designed using IS: 10262-2009 and mix proportions for M60 and M80 grade concrete are designed using Entroy and Shacklock’s Empirical Graphs. The mix proportions are given in Table 1.

Microorganism Used

Bacillus Subtilis JC3

Aerobic alkaliphilic spore-forming microorganism Bacillus subtilis strain with accession number JC3 is a laboratory cultured soil bacterium, isolated, deposited, cultured and grown in distilled water at Bacteria Discovery Laboratory (JNTUH Hyderabad) as shown in Fig. 1. This microor- ganism is used in preparation of bacterial concrete. Dif- ferent cell concentrations of bacteria were validated using serial dilution method. A member of the genus Bacillus, B. subtilis is rod-shaped, and has the capability to form a sturdy, protective endospore, allowing the organism to endure extreme environmental conditions. Bacillus subtilis cells naturally found in soil and vegetation are non- pathogenic and grows in the mesophilic temperature range of 25–35 C. The Bacillus subtilis JC3 strain isolated from soil has characteristics of incessant precipitation of dense insoluble calcite crystals and has high negative zeta-po- tential. Peptone: 5 gm/l., CaCl: 10 gm/l., NaCl: 5 gm/l.,

Yeast extract: 3 gm/l, Beef extract: 1.5 gm/l. Yeast and beef extracts are nitrogen source for bacteria growth. Chemical analysis on 10 5 cells/ml bacteria suspended nutrient based solution is done to test for its suitability as mixing water in concrete and results found are within the Permissible limits as per IS 456-2000.

Methodology

Two types of concrete mix of various grades i.e. one without bacteria (control sample) and one with bacteria (bacterial sample) were prepared for experimental inves- tigations. The mixes have similar proportions of sand, coarse aggregates, cement and water as per mix proportions shown in Table 1. However, the main difference was in the water component: (1) tap water is used for concrete spec- imens without bacteria (for M20, M40, M60 and M80 grades), (2) while Bacillus subtilis JC3 suspended water is used for specimens with bacteria (for M20B, M40B, M60B and M80B grades). For preparation of bacteria-nutrient suspension, optimum cell concentration of bacteria (Bacillus Subtilis JC3) is grown in nutrient based distilled water in sterile conditions at biotechnology laboratory. The bacterial culture in suspension is added to concrete during concrete mixing process. Concrete shall be mixed in a mechanical mixer.

Strength Studies

Experimental Investigations are carried out to evaluate the effectiveness of microbiologically induced mineral pre- cipitation on the mechanical properties of bacteria incor- porated concrete specimens. Brief summaries of these investigations are presented below.

123

Author's personal copy

J. Inst. Eng. India Ser. A

Optimum Cell Concentration for Promotion of Maximum Mineral Precipitation

To study the effect of cell concentration on the properties of concrete, bacteria incorporated cement mortar cubes of

size 70.6 9 70.6 9 70.6 mm were cast using

different

cell concentrations of Bacillus subtilis JC3 (1 9 10 4 cells/ ml, 1 9 10 5 cells/ml, 1 9 10 6 cells/ml, 1 9 10 7 cells/ml) and tested as per IS 12269:1987. For each cell concen- tration, six cubes were casted. This investigation was carried out primarily to understand the effect of bacterial cell concentration on the quantity of calcium carbonate precipitation. More bacteria with enough nutrients will precipitate more calcite in the laboratory conditions but in the cement-sand environment, bacteria mineral precipi- tating ability depends on its compatibility with cement- sand matrix. Bacteria incorporated into cement-sand medium should not affect the physio-chemical properties of cement-sand. Hence cell concentration of bacteria plays a key role in optimizing the performance of cementitious materials. The appropriate bacterial cell concentration for maximum calcium carbonate precipita- tion can be established by determining the 28 day com- pressive strengths of various cement-mortar specimens induced with different bacterial cell concentrations. The sample whose 28 day compressive strength was highest determines the optimum cell concentration for high amount of crystalline calcite precipitation. The cubes

having 1 9 10 5 cells/ml are found to have maximum strength. Henceforth, Optimum bacterial cell concentra- tion of 1 9 10 5 cells per ml of mixing water is used for further research. This improvement in compressive strength was mainly due to metabolic deposition of CaCO 3 in the voids or pores within cement–sand matrix modifying the pore structure of bacteria induced cement mortar specimens. During bacterial growth, the calcium precipitation process occurs continuously, clogging the internal pores with calcium precipitate. It is observed that for cement mortar specimens incorporated with Bacillus subtilis JC3 of cell concentration more than 10 5 cells/ml, the compressive strength is reduced. The gradual reduction of compressive strength of cement mortar cube specimens induced with bacterial cell concentrations more than 10 5 cells/ml of mixing water is attributed to the disruption of cement- mortar matrix integrity by the presence of organic matter (biomass) above the permissible limits as specified by IS:

3025-1986 and IS: 456-2000 as shown in Table 2. Max- imum strength is observed for 10 5 cells/ml of mixing water so this is taken as optimum cell concentration to be used for further study to investigate the effect of bacteria on properties of concrete.

Table 2 Properties of Bacterial suspension

Parameter

Results

Limits as per IS 456-2000 permissible limits (max)

pH

8.3

6.5–8.5

Chlorides, mg/l

145

2000 (PCC)

 

500 (RCC)

Alkalinity, ml

13

\ 25

Sulphates, mg/l

245

400

Fluorides, mg/l

0.022

1.5

Organic solids, mg/l

31

200

Inorganic solids, mg/l

1380.4

3000

Suspended matter, mg/l

1359

2000

Compressive, Split Tensile and Flexural Strength

To study the effect of biomineralization, due to Bacillus subtilis JC3 on the compressive, split tensile and flexural strength of concrete treated with 1 9 10 5 cells/ml bacteria spore suspension. For each grade, six cubes of size

  • 100 9 100 9 100 mm are cast and tested for compressive

strength as per IS 516:1959, six cylinders of size

  • 150 9 300 mm are cast and tested for split tensile strength

as per IS 5816:1999 and six prisms of size 100 9 500 mm are cast and tested for flexural strength as per IS 516:1959.

Stress–Strain Behaviour

In this study for each grade, six cylinder specimens of size

  • 150 9 300 mm are cast and are tested under strain control

under uni-axial compression as per IS: 516-1999 to get the stress–strain characteristics. From which peak stress, strain at peak stress and modulus of elasticity of bacterial con- crete specimens are determined.

Ultrasonic Pulse Velocity Measurements

In this study for each grade, bacteria treated concrete specimens strength and quality is assessed using non-de- structive testing by Ultrasonic Pulse Velocity on six cube specimens of size 150 9 150 9 150 mm as per IS: 13311 (Part 1): 1992.

Durability Studies

The understand the durability performance of bacterial concrete, the study of key parameters such as water absorption capacity, porosity, sorptivity, water permeabil- ity and chloride-ion penetration resistance studies are important. Brief summaries of these investigations are presented.

123

Author's personal copy

J. Inst. Eng. India Ser. A

Water Absorption Capacity (WAC)

In this study for each grade, the maximum quantity of water absorbed (WAC) by bacteria treated concrete spec- imens (six cube size 100 9 100 9 100) at room tempera- ture and pressure under conditions of saturation expressed as a percentage of the dry mass of the sample is evaluated as per ASTM C642.

Porosity

In this study for each grade, percentage of voids present in the bacterial specimens of six size 100 9 100 mm cubes, cast as per DIN 1048, were evaluated to understand the phenomena of water transport within pore structure and estimate the interconnected pore space. Porosity; g ¼ V v =q w V ¼ W sat W dry =q w V ¼ W w =q w V

where, V v = volume of voids in cc = W sat - W dry; V = total volume of specimen; q w the unit mass of water; W sat = weight of saturated specimen; W dry = weight of oven dried specimen (105 C).

Sorptivity

Sorptivity measures the rate of penetration of water into the pores in concrete by capillary suction. As per ASTM C1585, for each grade, the test for sorptivity was conducted on six numbers of 100 9 100 9 100 mm cubes by immersing them to a depth of 5–10 mm in the water and then measuring the gain in mass at regular intervals of 30 min for 72 h until there is no gain in weight. The cumulative volume of water that has pen- etrated per unit surface area of exposure ‘Q/A’ is plotted against the square root of time of exposure ‘ Ht’. The resulting graph could be approximated by a straight line passing through the origin. The slope of this straight line gives sorptivity coefficient ‘k’ which is considered as a measure of rate of movement of water through the capillary pores.

Water Permeability

For each grade, the tests for permeability were carried out on three 100 mm 9 100 mm cylinders using Concrete permeability apparatus as per IS: 3085-1965. The total quantity of water percolated through the cylinder speci- mens was collected in a glass bottle for a period of 100 h using which coefficient of permeability was calculated.

Chloride-Ion Penetration Resistance

To establish the resistance of bacterial concrete to chloride ion penetration, AASHTO T277 (ASTM C 1202) ‘Standard Test Method for Electrical Indication of Concrete’s Ability to Resist Chloride Ion Penetration’ was conducted and experimental investigations are presented. In this test, six concrete specimens of water-saturated, 50-mm thick, 100-mm diameter is subjected to a 60 V applied DC voltage for 6 h for each grade.

SEM Investigations

Microbial calcite precipitation was visualized by Scanning electron micrographs

Test Results and Discussions

The test results of Strength, Durability characteristics of bacterial concrete are listed in Tables 3, 4 and 5. The results on strength and durability characteristics are sum- marized as follows:

  • 1. For cell concentration of 10 5 cells/ml of mixing water the compressive strength of bacteria incorporated cement mortar specimens is maximum (increased by about 17%) at age 28 days as shown in Table 1 and Fig. 2. Chemical analysis on 1 9 10 4 cells/ml, 1 9 10 5 cells/ml, 1 9 10 6 cells/ml, 1 9 10 7 cells/ ml bacteria suspended nutrient based solutions are done to test to determine the presence of impurities. Suitability as mixing water in concrete depends on

Table 3 Effect of Bacillus subtilis JC3 JC3 cell concentration on compressive strength (MPa)

Cell concentration/ml of mixing water

Effect of optimum cell concentration on compressive strength, MPa

7 days

% Increase

14 days

% Increase

28 days

% Increase

Nil (control)

10

4

10

5

10

6

10

7

37.32

41.68

45.02

43.09

40.11

11.68

20.63

15.46

7.48

44.1

45.23

49.21

47.69

45.97

2.56

11.59

8.14

4.24

51.81

58.02

61.79

57.21

54.66

11.99

16.15

10.42

5.51

Author's personal copy

J. Inst. Eng. India Ser. A

Table 4 Strength characteristics of controlled and Bacterial concrete mixes at various ages

Strength studies

Grade ?

Controlled concrete

Bacterial concrete

M20

M40

M60

M80

M20

M40

M60

M80

Age

Compressive strength, MPa

 

28 days

28.18

52.01

72.61

93.8

32.74

61.06

94.21

119.2

60 days

32.44

56.47

79.26

98.35

37.97

64.92

102.9

125.6

90 days

33.27

57.96

83.59

107.57

40.4

66.83

108.6

138.14

Split tensile strength, MPa

 

28 days

3.26

4.51

4.63

4.88

3.73

5.13

5.63

5.76

60 days

3.34

4.63

4.71

4.93

3.89

5.41

5.82

6.02

90 days

3.49

4.89

4.79

5.07

4.04

5.50

5.93

6.12

Flexural strength, MPa

 

28 days

4.68

6.11

8.64

9.40

6.11

7.73

10.44

11.72

60 days

4.93

6.46

8.66

9.48

6.32

8.06

10.91

11.88

90 days

5.12

6.83

9.08

9.56

6.61

8.56

11.23

11.96

Ultrasonic pulse velocity, km/s

 

28 days

4.26

4.49

4.89

5.13

4.77

4.93

5.22

5.94

60 days

4.36

4.53

4.92

5.19

4.83

4.99

5.36

6.02

90 days

4.41

4.61

4.99

5.33

4.91

5.02

5.41

6.05

Modulus of elasticity, GPa

 

28 days

22.4

32.2

38.7

43.6

27.2

36.7

44.1

50.9

Peak stress, MPa

28 days

24.25

42.46

72.61

98.50

28.31

48.12

94.21

113.00

Strain at peak stress

 

28 days

0.0020

0.0021

0.0023

0.0020

0.0021

0.0021

0.0023

0.0024

the presence of impurities within permissible limits as per IS: 3025-1986 and IS: 456-2000 as shown in Table 3. An increase in bacterial cell concentrations above 1 9 10 5 cells/ml of mixing water reduces the strength of cement mortar cube specimens due to disruption of cement mortar matrix integrity with the presence of organic matter in the form of biomass, above the permissible limits specified as per IS:

3025-1986 and IS: 456-2000 in mixing water.

  • 2. It was observed that with the addition of bacteria, the Compressive, Split Tensile and Flexural Strength of concrete showed significant increase by nearly 25, 20 and 25% respectively in all grades of concrete proposed for all ages considered as shown in Table 4. Higher the grade more increase was noted because the voids and pores are sealed up by mineral precipitation due to bacteria. Reduction in pores due to such material growth has obviously increased the strength characteristics of concrete. Figure 3 shows variation of compressive strength of controlled and bacterial concretes of various grades.

  • 3. The relationship between stress and strain is impor- tant in understanding the basic elastic behaviour of

concrete in hardened state. From the values of stresses and strains, average stress–strain curves for different grades of bacterial concretes are plotted. The observation made from stress–strain curves is all grades of bacterial concrete have shown improved stress values for the same strain levels compared to that of conventional concrete mixes. Ultimate stress, strain at ultimate stress and modulus of elasticity (E) are determined from the stress–strain plots as shown in Fig. 4 and are listed in Table 4. The increase in E value is nearly 20–30% for low to high grade bacterial concrete.

  • 4. Ultrasonic Pulse Velocity test conducted on bacterial concrete specimens revealed that ultrasonic pulse velocity values increased with bacteria induction into concrete due to enhanced pore structure and microstructure of hardened bacterial concrete of all grades. The ultrasonic pulse velocity values are listed in Table 4. The pulse velocity values obtained for bacteria induced concrete were greater than 4.5 km/s which classifies as excellent concretes in terms of strength and durability point of view. From the results it is observed that the reduction in pores in

123

Author's personal copy

J. Inst. Eng. India Ser. A

Table 5 Durability characteristics of controlled and Bacterial concrete mixes at various ages

Durability studies

 

Controlled concrete

 

Bacterial concrete

Grade ?

M20

M40

M60

M80

M20

M40

M60

M80

 

Water absorption capacity, %

 

28 days

5.6

2.8

1.9

0.8

2.8

1.2

0.8

0.4

60 days

5.1

2.6

1.6

0.7

2.5

1.0

0.7

0.3

90 days

4.7

2.1

1.2

0.6

2.1

0.9

0.6

0.2

Porosity, %

28 days

3.65

3.22

2.35

2.22

1.09

1.02

0.93

0.87

60 days

3.44

3.03

2.11

1.98

1.01

0.92

0.88

0.82

90 days

3.41

2.97

1.96

1.72

0.95

0.88

0.8

0.77

Sorptivity, mm/min 0.5

 

28 days

0.124

0.092

0.070

0.055

0.091

0.071

0.052

0.041

60 days

0.109

0.083

0.053

0.041

0.074

0.062

0.049

0.032

90 days

0.097

0.074

0.044

0.036

0.055

0.054

0.041

0.022

Water permeability (depth of penetration, mm)

 

28 days

23 17 955421

 

60 days

23

17

9

5

5

4

NIL

NIL

90 days

20

15

7

3

3

3

NIL

NIL

Rapid chloride penetration (charge passed, coulombs)

 

28 days

2419

2008

1022

986

367

238

173

141

60 days

2213

1991

997

933

351

222

159

127

90 days

2100

1817

943

890

327

202

96

85

Author's personal copy J. Inst. Eng. India Ser. A Table 5 Durability characteristics of controlled and

Fig. 2 Percentage increase of compressive strengths at various Bacteria cell concentrations

bacterial concrete improves the surface integrity of concrete, improves its homogeneity good bonding and reduces the probability of cracks. Similar obser- vation is noted in all the grades of bacteria treated concrete specimens. So the ultrasonic pulse velocity measurements are obtained to assess particle conti- nuity inside the concrete, density and modulus of elasticity can be understood.

  • 5. Table 5 shows the WAC of controlled and bacteria incorporated concrete specimens of different grades

at various ages considered. Water Absorption Capac- ity (WAC) of bacterial concrete specimens is reduced by nearly 50–80% for low to high grade concretes as compared with WAC in controlled concrete speci- mens due to pore plugging with bacteria produced minerals thereby modifying the pore structure of the cement –sand matrix. The absorption characteristics indirectly represent the volume of pores and their connectivity. Figure 5 generalizes the amount of water absorption with time for different grades of controlled and bacterial specimens.

  • 6. Table 5 lists the porosity of controlled and bacteria incorporated concrete specimens of different grades at various ages considered. Porosity of concrete specimens is reduced by nearly 70% with induction of bacteria into concrete. The possible reason for this is calcite mineral precipitation in the pores reduced the average pore radius of concrete.

  • 7. Table 5 lists the sorptivity values of controlled and bacteria incorporated concrete specimens of different grades at various ages. The high sorptivity values are obtained for controlled concrete than bacterial con- crete. Sorptivity values for bacterial induced con- cretes were in the range of 0.091 to 0.041 mm/min 0.5 for ordinary to high grades and for control concretes

123

Author's personal copy

J. Inst. Eng. India Ser. A

Fig. 3 Strength development of a normal concrete and Bacterial concrete

Author's personal copy J. Inst. Eng. India Ser. A Fig. 3 Strength development of a normal

Fig. 4 Stress–Strain curves of controlled and Bacterial concretes of different grades

Author's personal copy J. Inst. Eng. India Ser. A Fig. 3 Strength development of a normal

the value is in the range of 0.124 to 0.055 mm/min 0.5 for ordinary to high grades at 28 days.

  • 8. Very negligible percolation of water has been found for higher grades of bacterial concrete specimens whereas for the ordinary and standard grades there is some percolation. Cylinders were removed from the permeability mould and were split to measure the water penetration depth. This confirmed that use of bacteria had resulted in almost impermeable con- crete. Table 5 lists the water penetration depths of controlled and bacteria incorporated concrete speci- mens of different grades at various ages and the

values are checked against the specifications sug- gested by MORT&H 4 th Revision Clause 1716.5. Results show that the presence of bacteria resulted in lower coefficient of permeability of range 0.23–0.27 9 10 - 9 m/sec in comparison to controlled concrete which has coefficient of permeability of 0.95–2.31 9 10 - 9 m/sec for various grades of con- cretes. Water permeability reduces in bacterial con- crete by nearly 70–90% from low to high grades suggesting that reduction in the permeability is due to reduction in the porosity of the concrete resulting from pore filling by calcite crystals.

123

Author's personal copy

J. Inst. Eng. India Ser. A

Fig. 5 Plot showing amount of water absorption with time

Author's personal copy J. Inst. Eng. India Ser. A Fig. 5 Plot showing amount of water
Author's personal copy J. Inst. Eng. India Ser. A Fig. 5 Plot showing amount of water

Fig. 6 a Cell Concentration–Nil (Control Specimen). b Cell Concentration—10 5 /ml (Optimum)

  • 9. The test results in Table 5 show that the addition of bacteria to concrete exhibit better chloride ion penetration resistance in all the grades for various ages considered. The electrical conductance of the test specimen, expressed as the total electrical charge passed through the specimen, in coulomb is observed to be very low in Bacterial Concrete. In case of high grade concretes, with bacteria induced in it, chloride ion permeability is reduced to negligible levels. Results indicates that bacteria induced concrete has shown between 85 and 90% significant improved resistance against the chloride movements in Bacte- rial Concrete as compared to the chloride movements

in normal concrete due to the good packing density between the particles. 10. Figure 4 shows Scanning Electron Micrographs (SEM) of cement mortar specimens showing dense calcite precipitation as calcite crystals with rod- shaped impressions housed by Bacillus subtilis. Bacteria promote calcium carbonate precipitation in the form of calcite crystals due to its metabolic reactions. The formation of calcite (CaCO 3 ) by process of bio-mineralization can be analyzed using various characterization techniques/methods. The micro-structural observations could improve the understanding of the mechanism of self-healing phenomenon by calcifying bacteria. Microbial calcite

123

Author's personal copy

J. Inst. Eng. India Ser. A

precipitation was visualized by SEM. Improvement in pore structure of cement-sand mortar samples treated with Bacillus subtilis JC3 of 10 5 cell concentration per ml can be observed in a magnified view (2500x) of SEM micrograph as shown in Fig. 6a, b.

Conclusion

Based on the results of investigation reported in this paper, the following conclusions are made-

The Compressive Strength, Split tensile Strength and Flexural Strength of concrete, incorporated with opti- mum cell concentration of bacteria, increases pro- foundly due to pore plugging by calcite mineral precipitation during microbial metabolic process of Bacillus subtilis JC3. The Bacterial concrete mixes have shown improved stress values for the same strain levels compared to that of controlled concrete mixes in low, medium and high strength grades resulting in the increase of elastic modulus. Bacteria treated concrete samples gave the lower water absorption, sorptivity and porosity values compared to control concrete. This means that the time taken for the water to rise by capillary action in bacterial concrete are longer and thus proved that these concrete are less porous compared to the control concrete. The possible reason for this is calcite mineral precipitation in the pores reduced the average pore radius of concrete by blocking the large voids (pore discontinuity) in the hydrated cement paste. Since interconnected pores are significant for permeability, the water permeability and chloride ion penetration are reduced relatively in bacteria treated specimens.

References

  • 1. H.M. Jonkers, A. Thijssen, Bacteria mediated remediation of concrete structures, in Proceedings of the Second International Symposium On Service Life Design for Infrastructures (Delft, The Netherlands, 2010)

  • 2. J.Y. Wang, K. Van Tittelboom, N. De Belie, W. Verstraete, Potential of applying bacteria to heal cracks in concrete, in Proceedings of the Second International Conference on Sus- tainable Construction Materials and Technologies, (Ancona, Italy 2010)

  • 3. W. De Muynck, D. Debrouwer, N. DeBelie, W. Verstraete, Bacterial carbonate precipitation improves the durability of cementitious materials. Cem. Concr. Res. 38, 1005–1014p (2008)

  • 4. H.M. Jonkers, Development of self-healing concrete: towards full scale application, in 3rd Internatinal Conference on Self-Healing Materials, (Bath, UK, 2011), p. 27

  • 5. S.K. Ramachandran, V. Ramakrishnan, S.S. Bang, Remediation of concrete using micro-organisms. ACI Mater. J. 98, 3–9 (2001)

  • 6. P. Ghosh, S. Mandal, B.D. Chattopadhyay, S. Pal, Use of microorganism to improve the strength of cement mortar. Cem. Concr. Res. 35(10), 1980–1983 (2005)

  • 7. J. Shen, X. Cheng, Laboratory investigation on restoration of Chinese ancient masonry buildings using microbial carbonate precipitation, in 1st BioGeoCivil Engineering Conference (Delft, 2008)

  • 8. Q. Chunxiang, W. Jianyun, W. Ruixing, C. Liang, Corrosion protection of cement-based building materials by surface depo- sition of CaCO 3 by Bacillus pasteurii. Mater. Sci. Eng.: C 29(4):1273-1280 (2009)

  • 9. C. Qian, R. Wang, J. Wang, The method of producing CaCO 3 by bacterial biomineralization. ZL2005100947744.5 (2007)

  • 10. J.P. Adolphe, J.F. Loubie` re, J. Paradas, F. Soleilhavoup, Proce´ de´ de traitement biologique d’une surface artificielle. European patent 90400G97.0. (French patent 8903517, 1989) (1990)

  • 11. M. Shirakawa, M. Cincotto, C. Dias, D. Atencio, M.J. Vanderley, Influence of carbonation in accelerated chamber previous to biocalcification on fiber cement surface, in 1st BioGeoCivil Engineering Conference (Delft, 2008)

  • 12. M. Saroj, A process for preparing modified bioconcrete. C04B 28/02 (Indian patent) (2006)

  • 13. S.S. Bang, J.K. Galinat, V. Ramakrishnan, Calcite precipitation induced by polyurethane-immobilized Bacillus pasteurii. Enzyme Microb. Technol. 28(4–5):404–409 (2001)

123