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Bio-Inspired Solutions for Durable Concrete

V Srinivasa Reddy1, M V Seshagiri Rao2, Ch Sasikala3 1- Associate Professor, Department of Civil Engineering, GRIET, Hyderabad. 2- Professor, Department of Civil Engineering, JNTUH College of Engineering, Hyderabad. 3- Professor, Centre for Environment, JNTU, Hyderabad.

ABSTRACT Unlike man-made materials, natural materials can interact with the surrounding environment, protect themselves from it, and heal autonomously when damage occurs. Inspired by such properties of natural materials, research on the development of innovative and novel bacterial concrete is underway and few research findings on its durability characteristics were presented in this paper. The notion of self-healing concrete that can repair itself without human intervention seems to be the stuff of science fiction till now but research conducted at JNTU Hyderabad on bacteria incorporated concrete realizes the potential of this new field of bio-inspired self repair innovation to extend the lifetime and safety of concrete structures, reduces demand for expensive conventional crack repair methods, and allows self remediation in human inaccessible locations. Bacterial concrete has self-healing component in the form of bacterium Bacillus subtilis JC3 and nutrients that will heal the damage to restore the integrity of the concrete structure. This emerging technology provided a benefit by extending the periods at which maintenance had to be carried out, and reducing the damage it sustained, extending the life of the concrete structure. Developing a bio-based concrete that would work reliably in potentially extreme conditions and remain stable over the life of the material was also a challenge. In the present investigations, artificially cracked cubes are immersed in bacteria mediated nutrient suspension for 7 days to study the mechanism of bio-remediation. The cubes were examined for the variation in their compressive strength and inferred that compressive strength showed increase. Our results showed that the durability properties of bacterial concrete enhanced enormously in terms of permeability, porosity, resistance to acid attack, chloride ion penetration. Key words: bacterial concrete, biomineralization, MICP, Bacillus subtilis JC3, durability. 1. Introduction For crack repair, a variety of techniques are available but traditional repair systems i.e., synthetic polymers have a number of disadvantageous aspects such as different thermal expansion coefficient than concrete and also pose environmental and health hazards. Therefore, bacteriogenic calcium carbonate precipitation has been proposed as an alternative and environmental friendly method to enhance the performance of concrete1. Several of these repairing systems are organic coatings consisting of volatile organic compounds. The air polluting effect of these compounds during manufacturing and coating has lead to the development of new formulations such as inorganic coating materials made up of calciumsilicate compounds, which exhibit a composition similar to cement. Concrete is a composite material with coarse and fine aggregates embedded in a cement paste matrix. As such, the aggregate and the cement paste as well as the interfacial zone between them affect the mechanical behaviour and permeability, thus durability of concrete. The transport of aggressive

gases and/or liquids into concrete depends on its permeation characteristics. As the permeation of concrete decreases, its durability performance, in terms of physicochemical degradation, increases. In concrete, it is generally not the porosity but the pore structure that is essential in establishing the permeability. In addition to that, microcracks in the matrix may contribute significantly to the permeability. Fig. 1 shows the difference between porosity and permeability schematically, and it indicates that the connectivity of the pore system is a prerequisite for permeability (i.e. an open pore system). A material can be porous and still perform tight as long as the pores are not interconnected (i.e. closed pore system).

Fig.1. Schematic diagram to distinguish porosity and permeability (EuroLightCon, 1998) Several bacteria have the ability to precipitate calcium carbonate. These bacteria can be found in soil, sand, natural minerals etc. Jonkers et al. used Bacillus cohnii bacteria to precipitate CaCO3. Bacillus pasteurii have been used by Santhosh et al.2, Day et al.2, Bang et al.2 and Ramakrishnan et al2. , while Dick et al.3 used Bacillus lentus and Bacillus sphaericus. In the present study the enhanced performance potential of bacteria B.subtilis is reported. De Muynck et al. found that the influence of the calcium source is limited to the morphology of the crystals. By means of SEM observations, they proved that the presence of chloride ions resulted in rhombohedral crystals, while the presence of acetate ions resulted in spherical crystals4. Bacteria-based self-healing sustainable concrete which can extend its service life is our focus of research. Calcite precipitation due to microbial chemical process by specific alkali resistant microorganisms can act as a self healing agent when induced into concrete. This mechanism is of great interest for repair in concrete structures without human intervention5. The research findings shows that a new type of alkaliphilic aerobic microorganism belonging to Bacillus species, which when added to concrete enhances the strength and durability characteristics of concrete structures significantly due to growth of filler material called calcite(CaCO3) within the pores of the cementsand matrix leading to pore refinement and enhanced concrete microstructure6. This paper presents the research findings to suggest the potential use of the microbial calcite precipitation process in remediation of the surface cracks and subsurface of porous media. 2. Chemistry of biogenic activity by Bacillus Subtilis JC3 Calcium chloride was used for precipitation of calcium carbonate, while culture medium consisting of Peptone: 5 g/lit., NaCl: 5 g/lit., Yeast extract: 3 g/lit. and beef extract was used for cultivation of microorganisms. B.subtilis cell can attract Ca ions (Ca2+), which react with carbonate ions CO32- originating from peptone during oxidative deamination of amino acids. Simultaneously, ammonia ions NH4+ increase pH value in surrounding medium which improves calcite precipitation efficiency. Precipitation of calcium carbonate crystals occurs by

heterogeneous nucleation on the bacterial cell wall once super-saturation is achieved . The microbial precipitation of CaCO3 is determined by several factors including: the concentration of dissolved inorganic carbon, the pH, the concentration of calcium ions and the presence of nucleation sites7. The first three factors are provided by the metabolism of the bacteria while the cell wall of the bacteria will act as a nucleation site8. This B.subtilis strain was chosen because of its greatest potential for precipitation of calcium carbonate, even under extreme conditions, as well as its lack of pathogenicity and a very negative zeta-potential9. The chemical equations involved in microbial activity are: Ca2+ + B.subtilis Cell B.subtilis Cell- Ca2+ CH3CH(NH2)COOH (Peptone) + O2 ---------> C2H2 + H2CO3 + NH3 H2CO3 ----------> H+ + HCO3NH3 + H2O --------> NH4+ + OHB.subtilis Cell- Ca2+ + CO32- B.subtilis Cell- CaCO3 3. Experimental Investigations 3.1 Compressive Strength To study the effect of biomineralization, due to Bacillus subtilis JC3 on the compressive strength of concrete treated with 1x 105 cells/ml bacteria spore suspension, Cubes of size 100 x 100 x 100 mm are cast and tested for compressive strength as per IS 516:1959. 3.2 Water Absorption The water absorption test was conducted as per ASTM C 642 (82) in order to determine the increase in resistance towards water penetration in concrete. The cube specimens of 100x100mm were prepared both with and without bacteria. After28 days curing, the specimens were oven dried at 105 C in oven, establishing a mass equilibrium of less than 0.5% between two measurements at 24 h intervals. Then the specimens were immersed in water at approximately 21 C for 48 h and saturated mass after immersion was calculated. Then the specimens were placed in suitable receptacle, covered with tap water and were boiled for 5 h, further the saturated mass after boiling was calculated. The specimens were suspended by a wire and the apparent mass in water was calculated. The ratio of the difference between the mass of saturated surface dry specimen and the mass of the oven dried specimen to the volume of the specimen (1000 ml) gives the permeable voids in percentage. Volume of permeable voids %= (C-A)/(C-D) x 100 where A is the mass of the oven dried sample in air (gm) C is the mass of sample after immersion and boiling (gm) and D is the apparent mass of sample in water after immersion and boiling ( gm). The oven dried cubes after attaining constant weight, were then immersed in water and the weight gain was measured at regular intervals until a constant weight was reached. The absorption at 30 min (initial surface absorption) and final absorption was determined. The final absorption in all cases was determined at 96 hrs. The absorption characteristics indirectly represent the volume of pores and their connectivity. 3.3 Sorptivity Test as per ASTM C1585 Sorptivity is a measure of the capillary forces exerted by the pore structure causing fluids to be drawn in to the body of the material. It provides a relative measure that combines pore size diameter and number of pores. The samples (100 x 50 mm size cylindrical specimens) are preconditioned to a certain moisture condition, either by drying the sample for 7 days in a 50C

oven. The sides of the concrete sample are sealed, typically with electricians tape or by sealant while the suction face and the face opposite it were left unsealed. The initial mass of the sample is taken and at time 0, is immersed to a depth of 5-10 mm in the water. The procedure was repeated, consecutively, at various times such as 15 min, 30 min, 1 hr, 2 hr, 4 hr, 6 hr, 24 hr, 48 hr and 72 hr until the last reading. The gain in mass per unit area over the density of water (I) is plotted versus the square root of the elapsed time (t). The slope of the line of best fit of these points (ignoring the origin) is reported as the sorptivity (k). For one dimensional flow, it can be stated that : I = k x t Where k is sorptivity coefficient and I = W/(A x d) W = the amount of water absorbed in kg ; A= the cross-section of the specimen that was in contact with water (m2) ; d= density of the medium in which the specimen was dipped (1000 kg/m3 in case medium is water). The rate of water absorption, sorptivity (k), is the slope of I- t graph (m / min1/2 or kg/ m2 / min). Because of small initial surface tension and buoyancy effects, the relationship between cumulative water absorption (kg/m2) and square root of exposure time (t 0.5) shows deviation from linearity during first few minutes. Thus, for the calculation of sorptivity coefficient, only the section of the curves for exposure period from 15 min to 72 hrs, where the curves were consistently linear, was used for the calculation of Sorptivity. While volume of permeable voids (VPV) is a measure of voids, Sorptivity gives a measure of nominal pore radius and number of pores. 3.4 Brenauer-Emmett-Tellers (BET) Nitrogen (N2) Nitrogen Adsorption Method BET nitrogen adsorption method is used to determine the volume, size and surface area of pores. Porosity characteristics of Concrete including total volume of pores in concrete matrix, volume of micropores, pore size distribution and specific surface area. This test confirms the modification in pore size distribution due to the addition of microorganisms. The porosity and pore size distribution as measured by BET Nitrogen Adsorption Method are reported graphically as shown in Fig 7. 3.5 Thermogravimetric Analysis A sample of the repair material was removed from the crack by means of a needle. This material was used for thermogravimetric analysis (TGA). About 30 mg of the repair material was brought in a sample cup. Subsequently the cup was placed in the TGA apparatus. During the analysis, the crack repair material was exposed to temperatures ranging from 20 C to 900 C at a rate of 10 C/min in an inert argon atmosphere. Through performance of the TGA analysis, the presence of CaCO3 in the repair material is determined. When CaCO3 crystals are present in the repair material, they will decompose into CaO and CO2 upon heating. As CaCO3 decomposes between the temperature ranges of 500-800 C a decrease in weight, caused by the release of CO2, is expected around that temperature interval for the samples treated with bacteria. As it can be seen in Fig 7, at about 100 -200 C the water in the specimens evaporates, leading to a decrease in weight. Between 500 and 800 C another decrease in weight is observed due to the decomposition of CaCO3. For the without bacteria specimens the weight loss is rather small while for with bacteria specimens a strong decrease in weight is observed. These results provide evidence that particularly in the case of concrete specimens with bacteria, CaCO3 crystals are formed. In other graph, the percentage change in weight / C is shown versus the temperature to indicate the points at which the weight loss is most apparent. For concrete specimens without bacteria only a small amount of CaCO3 is decomposed at the temperature of 591 C. This decrease in weight may possibly be attributed to chemical precipitation of CaCO3. For concrete

specimens with bacteria, the observed peaks are more distinct. XRD analysis was performed to find out which type of material was formed besides CaCO3. It was shown that the crystalline materials present in the mortar matrix were calcite, vaterite and aragonite. All these are crystalline forms of CaCO3 from which calcite is the most stable form. Aragonite and vaterite undergo transitions to calcite at 455 C and between 350 and 400 C respectively. Consequently about 600 C the reactant CaCO3 would normally be the calcite. 4. Test Results and Discussion Test results of various investigations carried out on bacteria incorporated concrete are presented below: Table 1: Effect of Bacillus subtilis JC3 cell concentration on Compressive Strength (MPa) Average Compressive Strength (MPa) S.D % % Cell concentration/ml % Increase Increase Increase of mixing water 7 days relative to 14 days 28 days relative to relative to control control control Nil (control) 37.32 0.42 44.10.66 51.810.10 104 41.680.52 11.68 45.230.85 2.56 58.020.72 11.99 105 45.020.22 20.63 49.210.91 11.59 61.790.68 16.15 106 43.090.36 15.46 47.690.32 8.14 57.210.49 10.42 107 40.110.58 7.48 45.970.44 4.24 54.660.89 5.51 An alkaliphilic aerobic soil microorganism Bacillus subtilis JC3 is incorporated at different cell concentrations with the mixing water. Different cell concentrations were derived from the bacterial growth culture by serial dilution method. The study showed that a 16 % increase in 28 day compressive strength of cement mortar was achieved with the addition of about 105 cell/ml of mixing water as shown in Table 1. The strength improvement is due to growth of filler material within the pores of the cementsand matrix as shown in Fig 2. The modification in pore size distribution and total pore volume of cementsand mortar due to such growth is also noted.

Fig 2: Strength development of a Normal concrete and Bacterial concrete

Fig 3: Schematic drawing of Sorptivity Test setup Table 2: (a) Water Absorption studies on Specimens of age 28 days Water Absorption % at 30 min Grade of Concrete Ordinary grade concrete (M20) Standard grade concrete (M40) High Strength grade Concrete (M60) Controlled Concrete (without microorganism) 3.17 2.64 1.94 Bacterial Concrete (105 cells/ml of microorganisms) 1.76 1.14 0.78

(b) Acceptance Criteria for absorption Absorption (%) at 30 minutes < 3.0 3.0 to 5.0 > 5.0 Absorption rating low average high Concrete quality good average poor

(c) Water Absorption Capacity (WAC) (%) and Volume of permeable voids (VPV) (%) Controlled Concrete Bacterial Concrete M20 M40 M60 M80 M20 M40 M60 M80 Water Absorption Capacity (WAC) (%) 5.62 2.79 1.93 1.52 2.79 1.19 0.77 0.38 12 6 4 3 6 3 2 1 Volume of permeable voids (VPV) (%) It was observed that with the inclusion of bacteria, water absorption capacity of concrete decreased. Maximum reduction in water absorption was observed with 105 cells/ml for bacterial concrete specimens as shown in Table 2 a-c. The deposition of a layer of calcium carbonate on the surface and inside pores of the concrete specimens resulted in a decrease of water absorption and permeability as shown in Fig 4 and 5. Once the pores are sealed, reduction in water ingress is observed. This bacterial action deposition can seal the pores, voids and microcracks, where other sealants are unable to work through. Concrete specimens treated with bacteria and nutrients

showed significantly less water absorption compared to untreated specimens indicating that about 65% of this decrease was attributable to the presence of biological matter, which physically hindered the movement of water as shown in Fig 6. This improvement is probably due to deposition on the microorganism cell surfaces and within the pores of cementsand matrix, which plug the pores within the mortar. As a consequence, the carbonation rate decreased by about 2530%.

Fig 4: Water Absorption Capacity (WAC) (%)

Fig 5: Volume of permeable voids (VPV) (%)

0.009 M20 Controlled 0.008 M40 Controlled M60 Controlled M80 Controlled 0.007 M20 Bacterial M40 Bacterial M60 Bacterial 0.006 M80 Bacterial

I x 10 -3 m






0 0.00



30.00 t0.5 min





Fig 6: Plot between I- t to calculate sorptivity coefficient (k) A significant decrease in porosity and average pore diameter was observed in bacterial concrete by the addition of bacteria as compared to the control concrete as shown in Fig 7. Efficiency of this bacteriogenic mineral plugging was also evaluated by means of ultrasonic transmission measurements and visual examination. The presence of CaCO3 in the concrete is determined TGA analysis. The CaCO3 crystals present will decompose into CaO and CO2 upon heating (CaCO3 CaO + CO2). As CaCO3 decomposes between the temperature range of 500700C a significant decrease in weight, caused by the release of CO2, is expected around that temperature interval for the samples treated with bacteria. A clear difference in weight loss can be observed between the samples treated with and without bacteria as shown in Fig 8.

Cumulative Pore Volume (cc/g)

M20 Controlled


M40 Controlled M60 Controlled M80 Controlled


4.00E-03 2.00E-03

0.00E+00 0 5 10 15 20 25 30 35 Pore Diameter (nm)

Fig 7: Pore volume versus Pore Diameter

105 without bacteria with bacteria 100

Weight %




75 0 100 200 300 400 500 Temperature C

expulsion of evaporable water hydratation of the decomposition of hydrate calcium Ca(OH) 2 silicate decomposition of hydrate calcium silicate






CaCO3 Disintegrates

0.4 without bacteria with bacteria 0.3

Change in Weight (%/ oC)



0 0 100 200 300 400 500







Temperature C

Fig 8: TGA Results showing weight loss and Change in weight loss per oC

5. Conclusions In bacterial concrete, induction of microorganisms inside the concrete has enormous effect on the porosity within the cement matrix paste, on the particle size distribution of the crystalline phases and on the presence of in-homogeneities within the hydrated paste due to mineral precipitation. Calcite mineral precipitation results in less capillary porosity in the hardened paste and hence a greater strength. This reduced capillary porosity also favours the formation of finetextured hydration products with optimized particle size distribution of the cementitious materials in order to increase the potential packing density. So bacteria incorporated concrete has increased packing density and reduced capillary porosity. The calcite crystals formed will glue together the hydrated particles which reduce the interstitial porosity between them. Furthermore the effect of mineral precipitation homogenously in bacterial concrete leads to a reduction in inhomogeneities within the paste and hence improved paste strength. The strength of the paste will be limited by the flaws that form the weakest link, be the inhomogeneities or capillary pores. In order to improve the strength of the paste as a whole, all such flaws must be minimized. Therefore bacterial concrete is a new approach to enhance the strength and durability of the concrete economically. From the investigation, it has been revealed that bacterial concrete has better resistance against strength deterioration for all curing conditions and curing ages. From the above, it is clear that the presence of a layer of carbonate crystals on the surface has the potential to improve the resistance of cementitious materials towards degradation processes. The use of bacteria in concrete mix also needs further research efforts. Several issues still need to be addressed in this field: (a) Which calcite producing bacteria are more efficient in highly alkaline environment? (b) Which is the most eco-efficient encapsulation method? (c) Will biologically deposited calcite endure the test of time? (d) Can biomineralization be made cost-efficient? (e) What are the environmental implications related to the use of corn steep liquor as a nutrient source? (f) Are there any health implications involved in the use of bacteria? (g) What is the life cycle analysis of biotech concrete? 6. References 1. S.K. Ramakrishnan, R.K. Panchalan, S.S. Bang, Improvement of concrete durability by bacterial mineral precipitation, Proceedings of 11th International Conference on Fracture, Turin, Italy, 2001. 2. S.K. Ramachandran, V. Ramakrishnan, S.S. Bang, Remediation of concrete using microorganisms, ACI Materials journal 98 (2001) 39. 3. P. Ghosh, S. Mandal, B.D. Chattopadhyay, S. Pal, Use of microorganism to improve the strength of cement mortar, Cement and Concrete Research 35 (10) (2005) 19801983. 4. J. Dick, W. De Windt, B. De Graef, H. Saveyn, P. Van der Meeren, N. De Belie, W. Verstraete, Bio-deposition of a calcium carbonate layer on degraded limestone by Bacillus species, Biodegradation V17 (4) (2006) 357367. 5. V.S. Whiffin, Microbial CaCO3 precipitation for the production of biocement, School of Biological Sciences and Biotechnology, Murdoch University, Perth, 2004, p. 155. 6.J. Morse (Ed.), The Kinetics of Calcium Carbonate Dissolution and Precipitation, Mineralogical Society of America, 1983.


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