MICROBIAL IMPROVEMENT

CEMENT MORTAR

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 CONTENTS
 Introduction

 Microbial Cement Mortar

 Preparation of Mortar

o Materials Used:

 Microorganisms

 Finding right bacteria

 Growth media and bacterial concentration

 Solutions for bacterial suspensions

 Mix proportion

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 Tests on Microbial Induced Cement Mortar
o Water absorption
o Compressive strength
o Scanning electron microscopy (SEM)
o Ultra sonic pulse velocity
o X-Ray Diffraction
 Case Study
 Advantages
 Disadvantages
 Conclusion
 Reference

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INTRODUCTION
 Industrialization and urbanization is facing the issues related to
new construction.

 Repair and rehabilitation is main concern for existing facilities.

 Concrete – a primary construction and repair material.

 A promising sustainable repair method- microbial improvement

cement mortar.

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MICROBIAL CEMENT MORTAR
 High performance construction material.
 Parameters involved in microbial concrete process:
 Calcite deposition
 pH increase
 Urea degrading capacity
 Extra cellular polymeric substances(EPS) production
 Biofilm formation

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PREPARATION OF MORTAR

 MATERIALS USED :
 Microorganisms
o Sporosarcina pastuerii
o Bacillus sphaericus
 Ordinay portland cement
 Standard sand
 Distilled water

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Microorganisms
• Bacteria obtained from :

 Microbial Type Culture Collection and Gene Bank

• First cultured in solid media (agar)

• Transferred to liquid media (nutrient broth)

• Kept in shaking incubator (for 48 hours)

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 Bacterial count :

 Concentration of cells – Haemocytometer

 Optical density – Spectrophotometer

 Gram staining :

 Determination of morphology of bacterial strains

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 Finding right bacteria

• Bacteria used :

 Sample of Endolithic bacteria and Bacteria in sediments were

used

 Genus Bacillus was found to thrive in high alkaline environment

 These bacteria produce spores – that have extremely thick cell

walls

 Bacteria become activated when the concrete starts to crack

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Table 1 : Various microbes used in concrete

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 Growth media and bacterial concentration

 S. Pasteurii –

 Ammonium sulphate and yeast extract medium:

 20g yeast extract

 10g di-ammonium sulphate

 20g agar

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  Nutrient broth medium:

 3g nutrient broth
 20g urea
 10g NH4Cl
 sodium bicarbonate

 E. Coli -
 Lysogeny broth medium

 10g tryptone

 5g yeast extract

 10g NaCl

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 Solutions for bacterial suspensions :

Table 2 :Composition of solutions used for preparing bacterial suspensions

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Mix Proportion
 Microorganisms of different cell concentration :
 104 cells/ml
 105cells/ml
 106 cells/ml
 107 cells/ml
 Cement to sand ratio - 1:3 (by weight)
 Water to cement ratio - 0.42.
 S. pasteurii correspondence to Optical Density(OD) at
600 nm of 0.5, 1.0 and 1.5 OD

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Mechanism
 Microbiologically induced calcite precipitation (MICT
method) is the mechanism.
 MICT method is also known bio mineralization.
 Living organisms produce minerals is referred to as
bio mineralization.
 These minerals include silicates, carbonates, calcium,
phosphates.
 Microbial activity has greater role in carbonate
formation.

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 Microbes are responsible for inducement of calcium
carbonate precipitate.
 Most evident minerals are carbonates
 Four characters are responsible for precipitation of
calcium carbonate.
1. Calcium concentration
2. Concentration of dissolved inorganic carbons
3. pH value
4. Availability of nucleation site.

 The minerals act as pore filling materials.

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 Carbonate precipitate generated microbially is mainly
act pore filling material.
 It restricting the flow of water and decreasing
permeability.
 Hence considerably increase in compressive strength.
 Calcium carbonate in the form of precipitation seal the
cracked surface on concrete as bone fractures.
 Crack is consumed during metabolic biochemical
reaction; as calcium carbonate helps in arresting
corrosion of steel in RCC works.
 Biologically induced cement based material also
exhibits better durability and crack repairing
performance compared to normal concrete material.

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TESTS ON MICROBIAL INDUCED
CEMENT MORTAR
 Water absorption :

 weighing the saturated specimens (W1) and dried specimens

in oven at 80 °C for 24 h (W2) at curing times of 3, 7, 14

and 28 days.

 Water absorption(%) = [(W2-W1)/W1]×100.

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 Compressive strength :

 Compressive strength measurements are carrying out using five

tones German Bruf Pressing Machine with a loading rate of 100

kg/min

 Scanning electron microscopy (SEM) :

 The scanning electron micrographs of freshly fractured specimens

were taken with Inspects (FEI Company, Holland) equipped with

an energy dispersive X-ray analyzer (EDAX) at the accelerating
voltage of 200 V to 30 kV

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 Ultra sonic pulse velocity :

 The time taken for the pulse to pass through the concrete is

measured by electronic Measuring circuits.

 X-Ray Diffraction:

 Reveals information about the crystal structure, chemical

composition and physical properties of bacterial cement

composites.

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 CASE STUDY
 Materials

 Microorganisms

 Bacteria species - Sporosarcina pasteurii (previously known

as B. pasteurii)

 obtained from National Collection of Industrial and

Marine Bacteria-England by national research center.

 The aerobic organisms is a moderately alkalophilic

(growth optimum pH 9.25).

 S. pasteurii. - ensures a high cell surface to volume ratio

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 Culture media
 Aerobic batch conditions in –

 10 g/l yeast extract

 5 g/l NaCl

 25 mM CaCl2

 20 g/l urea

 Medium pH adjusted before sterilization to 6.5 by 1 Normal HCl.

 Urea/ CaCl2 was added post autoclaving by 0.22 filter sterilization to

prevent chemical decomposition under autoclave condition

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  Cement and sand

 Ordinary portland cement (OPC) which provided by suez

cement company, el-suis governorate, egypt and sand
obtained from elwasta area, beni-suif governorate, egypt.

 Preparation of microbial cement mortar

 Sand: cement ratio - 3:1 by weight

 W/C ratio of 0.46

 1 inch cubic moulds were prepared.

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 Sand and cement were thoroughly mixed, adding along with grown
culture

 S. pasteurii correspondence to OD at 600 nm of 0.5, 1.0 and 1.5 OD;

bacterial cells concentration was measured by spectrophotometer at
wave length 600 nm.

 The fresh mortar pastes were cast into the mould and compacted on a
vibration machine then cured in humidity chamber with relative
humidity 100% for 24 h.

 After de-moulding the control specimens were cured under tap water
and the specimens with bacteria were cured under solution of 20 g/l
urea and 25 mM CaCl2 at room temperature until the times of testing
at the intervals of 3, 7, 14 and 28 days.
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 Results and discussion
 Water absorption:

 Water absorption of cement mortar (CM) specimens cured

under tap water and CM mixed with bacterial cell

concentrations of optical densities (0.5, 1 and 1.5 OD) cured
under growth media, at 3, 7, 14 and 28 days

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Fig 2 : Water absorption of cement mortar with and without bacterial cells up to 28 days.

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 Result :
 Water absorption of cement mortar with or without bacterial cells
decreases with time of curing up to 28 days - due to the
continuous hydration and accumulation of hydrated products
which fill the open pores of the specimens.
 The water absorption values of CM specimens mixed with
bacterial cells at all OD concentrations (0.5, 1, and 1.5) are lower
than those of control specimens.
 The water absorption values of cement mortar decrease with OD
of bacterial cells
 With increasing bacterial cells concentrations, the biomass and
precipitated calcite content increases which fill some of the open
pores and therefore, decreases the extent of water absorption.
 The water absorption is linearly proportional to the total porosity
of the cement mortar.
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 Compressive strength
 The compressive strength of CM specimens cured under tap
water and CM specimens mixed with bacterial cells and
cured under growth media.

Fig 3 :Compressive strength of cement mortar with and without

bacterial cells up to 28 days.
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 The strength values of CM specimens mixed with bacterial
cells are higher than those of control CM.
 During the initial curing period, microbial cells obtained good
nourishment, because the cement mortar was still porous
 The increase in compressive strength is mainly due to
consolidation of the CM matrix with a consequent filling of the
pores inside the CM cubes with microbiologically induced
calcium carbonate precipitation
 The compressive strength of CM increases with concentration
of bacterial cells up to 1 OD

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Fig 4 : Testing of cement mortar specimen:cube under compression test

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CONCLUSION
 The physico-mechanical properties, compressive
strength and water absorption, of the bacteria modified
mortar were improved due to the deposition of the new
calcite material by the bacterial activity.

 The water absorption of cement mortar decreases with

concentration of bacterial cells

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 The compressive strength increases with
concentration of bacterial cells up to 1 OD

 The optimum bacterial cells concentration which leads

to the highest improvement in mortar, such as higher
compressive strength and lower water absorption is 1
OD.

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Table 4 : Various construction Materials using MICP
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 Table 5 : Application of bacteria
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Fig 5 : Application of bacteria in construction area

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 ADVANTAGES
 Enhancement in compressive strength and stiffness values

 Remediation/ sealing of cracks in concrete

 Due to bacterial chemical process better resistant towards

freeze-thaw attack reduction
 Due to surface treatments reduction in gas permeability

 Arrest in corrosion due to consumption of oxygen ingress

through substances

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 By sealing the paths of ingress reduction in corrosion of
reinforced concrete

 Better life of concrete structures/ surfaces

 Reduction in water and chloride ion permeability

 Resistance to acid at pH greater than 1.5

 Eco friendly, antifungal, natural water purifier

 Restoration material for stone building/ monuments

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DISADVANTAGES
 Cost of bacterial concrete doubled in comparison to
conventional concrete

 Poor bacterial growth in any atmosphere and media

 No prescribed IS code in terms of doses of bacteria

for optimum performance

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 Costly investigation as calcite precipitation layers are
complex, requiring skilled staff

 Completion of bacterial activity a great concern

 concern Limited research in the field of MICP

 Lacks in substantial commercial applications

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CONCLUSION
 Conventional technology because of its being eco-friendly
and self healing.
 Improves the physical properties :
 Increases durability of various building materials

 Improves compressive strength

 Reduces permeability & water absorption

 Arrest the corrosion in steel

 Very convenient for usage

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 The physico- mechanical properties, compressive strength
and water absorption, of the bacteria modified mortar
were improved due to the deposition of the new calcite
material by the bacterial activity

 The strength and water absorption improvement are due to

the growth of calcite crystals within the pores of the
cement–sand matrix

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REFERENCE
1. H.Abbas, Q. S. Hadi, Saleh Alsayed Sheikh, Yousef Al-Sallom,
(2017). “Effect of some biotic factors on microbially-induced calcite
precipitation in cement mortar” Saudi Journal of Biological
Sciences(2017) 24, 286-294
2. Kenny Tiong Ping Chiet, Khairul Anuar Kassim, Murtala umar.
(2016) “Biological improvement of soil in civil engineering: A
review” Journal of Rock Mechanics and geotechnical engineering 8
(2016) 767-774.
3. Pratyush (2015): “Microbial concrete and Influence of Microbes on
Properties of Concrete” International Journal of Science and
Research (IJSR) 2319-7064 Index Copernicus Value (2015): 78.96.
4. A.H. Ali , Fatma N. Talkhan , H.A. Abdel-Gawwad, S.A. Abo-El-
Enein , (2013) “Application of microbial biocementation to improve
the physico-mechanical properties of cement mortar” HBRC Journal
(2013) 9, 36-40.

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 THANK YOU

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