PROPOSED PLAN OF RESEARCH WORK

EXPERIMENTAL EVALUATION OF MECHANICAL AND DURABILITY CHARACTERISTICS OF BACTERIA EMBEDDED CONCRETE

By V Srinivasa Reddy

Name & Designation of the suggested Supervisor Dr M V Seshagiri Rao Professor and Head Department of Civil Engineering Jawaharlal Nehru Technological University Hyderabad

RESEARCH This project proposal aims to study the

B!EC"#VE

echanical and Durability properties of concrete

embedded with bacteria and investigate the self! healing behavior of bacterial concrete in ordinary" standard and higher grades by embedding bacteria #Bacillus subtilis) for $ndian conditions% $n this research" $ propose to evaluate the effect of bacteria on the mechanical properties such as compressive strength" tensile strength" fle&ural strength" modulus of elasticity etc and on durability properties such as al'ali and sulfate attac' " shrin'age" deterioration studies$ abrasion resistance" air and water permeability" chloride permeability "salt scaling resistance of concrete and compare those properties between Concrete embedded with and without bacteria in ordinary" standard and higher grades and also investigate the self crac' remediation behavior of bacteria embedded concrete $ also propose to determine the suitability of Bacillus subtilis bacteria and its optimum concentration" its impact on various properties of concrete% No e&isting community research project is similar to my proposed wor'( therefore" this will be a big step in understanding the novel approach in concrete crac' remediation by microbiologically induced calcite precipitation% The objective of this proposal" therefore" is to study the Concrete embedded with bacteria #Bacillus subtilis) could be the one of the novel approaches in concrete crac' remediation% y proposed wor' will serve as a model for studying the effect of bacteria for improvement of concrete mechanical and durability characteristics by microbiologically inducing calcite mineral precipitates( this phenomenon is called as icrobiologically $nduced Calcite )recipitation # $C)*%

+lso the long term thermal effects on the bacteria induced concrete can be studied along with the cost effectiveness of bacterial concrete when compared to other self healing methods of concrete% y wor' will also ascertain the negative effects of bacteria if any and the effect of bacteria in concrete subjected to high temperatures% The rate of crac' healing and development patterns in the free,e -thaw cycles environment also can be studied%

BAC%&R 'ND Concrete" a strong" durable material composed of cement" aggregate and water" is the most widely used construction material" despite of its virility in construction" it is 'nown to have several limitations%

Concrete has an high load bearing capacity under compression" but is wea' in tension has limited ductility and little resistance to crac'ing% That is the reason concrete is reinforced with steel bars for the structures to carry tensile loads% The steel reinforced bar ta'es the load when the concrete fails in tension% +nother feature of concrete is to protect the steel reinforced bars from the environment and prevent corrosion% However" the crac's in the concrete are a major problem which affects the strength and durability of the structures% The ongoing research in the field of concrete technology has led to the development of special concrete considering #i* the speed of construction #ii* the strength of

concrete #iii* the durability of concrete and #iv* the environmental friendliness of industrial material li'e fly ash" blast furnace slag" silica fume" meta'eolin etc% .ecently" microbial mineral precipitation resulting from metabolic activities of favorable microorganisms in concrete improves the overall behaviors of concrete% Use of these /iomineralogy concepts in concrete leads to potential invention of new material called 0Ba(teria) Con(rete1% The ingress of water and chloride ions into the crac's deteriorates the structure by corroding the steel% To increase the strength and durability of the structure either the crac's that are formed should be repaired conventionally using epo&y injection" late& treatment etc or by providing e&tra reinforcement in the structure to ensure that the crac' width stays within a certain limit% Especially with current steel prices on steep rise" providing e&tra steel is not economically viable% ain reason to prevent crac's or

limit crac' widths is to enhance the durability of the structures% $f in some way a reliable method could be developed that repairs crac's in concrete automatically #self healing*" this would increase and ensure durability and functionality of structures enormously% +n $nnovative self healing biological approach is first proposed to ma'e use of bacterial concrete to heal crac's in concrete structures% 2o Concrete embedded with Bacillus subtilis could be the one of the novel approaches in concrete crac' remediation% The 0Ba(teria) Con(rete1 can be made by embedding bacteria in the concrete that are able to constantly precipitate calcite% This phenomenon is called microbiologically induced calcite precipitation # $C)*% $t has been shown that under favorable conditions " for instance Bacillus subtilis" a common soil bacterium" can continuously precipitate a new highly impermeable calcite layer over the surface of an already e&isting concrete layer% 3urther more the bacteria should be suspended in a determined optimi,ed concentration in a certain medium before they are mi&ed through the concrete ingredients% 4ptimi,ation of bacteria concentration is evaluated by conducting some e&perimental testing%

Detailed investigations carried out by 5% .ama'rishnan at 2outh Da'ota 2chool of

ines" 2outh

Da'ota" U%2%+%" have shown that bacteria can be used for improving the resistance of concrete to al'ali" sulphate attac'" drying shrin'age etc% which will increase the strength and durability of concrete% However" not much investigation is reported in $ndia for producing bacterial concrete using Bacillus subtilis% 6eeping this in view" the present e&perimental investigations are proposed to evaluate the mechanical and durability characteristics in ordinary grade concrete" standard grade of concrete and high grade concrete with and without addition of bacteria Bacillus subtilis%

BAC"ER#A* C NCRE"E The concept of bacterial concrete is first investigated in 7889 #5%.ama'rishnan et al.*% +n innovative approach in concrete crac' remediation utili,ing microbiologically induced calcite precipitation # $C)* is introduced% The $C) improves the impermeability of the concrete thus

increasing its resistance to al'aline" sulfate attac's and conse:uently increasing the strength and durability of concrete% + common soil bacterium" Bacillus subtilis" was used to induce CaC4; precipitation% The underlying process is that the microbial urease hydroly,es urea to produce ammonia and carbon dio&ide" and the ammonia released into surroundings subse:uently increases pH" leading to accumulation of insoluble CaC4;% .esearchers proposed different bacterial concretes with different types of bacteria cultures% The various bacteria used in the concrete are Bacillus Pasteurii" Bacillue Sphaericus, and E-coli etc% $n the present investigation an attempt is made by using the bacteria Bacillus subtilis% The main advantage of embedding Bacillus subtilis bacteria in the concrete is that it can constantly precipitate calcite% This phenomenon is called microbiologically induced calcite precipitation # $C)*% Calcium carbonate precipitation" a widespread phenomenon among bacteria" has been investigated due to its wide range of scientific and technological implications% Bacillus subtilis is a laboratory soil bacterium" which can produce calcite precipitates in suitable medium #urea buffer and CaCl 7* supplemented with a calcium source% This calcite layer improves the impermeability of the specimens thus increasing its resistance to al'aline" sulfate and free,e!thaw attac's and conse:uently increases the mechanical and durability properties of concrete%

RESEARCH ME"H D * &+

V, Rama-rishnan et al$ has proposed a novel techni:ue in remediation of crac's and fissures in concrete by microbiologically inducing calcite precipitation% icrobiologically induced calcite

precipitation is a techni:ue that comes under a broader category of science called biominerali,ation% Bacillus Pasteruii" a common soil bacterium can induce the precipitates of calcite% +s a microbial sealant" Calcite #CaC4;* e&hibited its positive potential in selectively consolidating simulated fractures and surface fissures in granites and in the consolidation of sand% $C) is highly desirable chemical

reaction because the calcite precipitation induced is a result of microbial activities% The techni:ue can be used to improve the compressive strength and stiffness of crac'ed concrete specimens% + durability study on concrete beams treated with bacteria" e&posed to al'aline" sulfate and free,e!thaw environments was studied by him% The effect of different concentrations of bacteria on the durability of concrete was also studied by him% $t was found that all the beams with bacteria performed better than the control beams #without bacteria*% The durability performance increased with increase in the concentration of bacteria% icrobial calcite precipitation was :uantified by <!ray diffraction #<.D*

analysis and visuali,ed by 2E % The uni:ue imaging and microanalysis capabilities of 2E established the presence of calcite precipitation inside crac's" rod shaped bacterial impressions and a new calcite layer on the surface of concrete% This calcite layer improves the impermeability of the specimen" thus increasing its resistance to al'aline" sulfate and free,e!thaw attac'%

/acterial deposition of a layer of calcite on the surface of the specimens resulted in a decrease of capillary water upta'e and permeability% This bacterial treatment resulted in a limited change of the chromatic aspect of mortar and concrete surfaces% The type of bacterial culture and medium composition had a profound impact on CaC4 ; crystal morphology% The use of pure cultures resulted in a more pronounced decrease in upta'e of water" respectively less pronounced change in the chromatic aspect" compared to the use of mi&ed ureolytic cultures as a paste%

E.PER#MEN"A* AC"#V#"#ES )rior studies have indicated that the bacterial concrete will have the higher life compared to conventional concrete of ordinary and standard grade% $n spite of its recent introduction the present

proposed study is ta'en up to study the mechanical and durability properties of high grade concretes% The proposed wor' focuses on three phases= PHASE / #0 Culture and >rowth of bacteria PHASE / ##0 To study the echanical properties of higher grade concrete such as compressive

strength" split tensile strength" fle&ural strength" modulus of elasticity" stress!strain behavior of concrete etc at normal temperatures and concrete subjected to elevated temperatures% PHASE / ###0 + concrete mi& developed in )hase-$$ is tested for durability aspects such as resistance to chemical attac' "corrosion of steel" shrin'age" strength loss" ?eight loss" +cid Durability 3actor "+cid +ttac' 3actor of concrete$ abrasion resistance" air and water permeability" chloride permeability "salt scaling resistance of concrete and to study the accelerated corrosion of crac'ing in concrete &ro1th of 2a(teria0 The first phase of investigation is carried out to culture of bacteria% The pure culture will be isolated from the soil sample of JNTU and will be maintained constantly on nutrient agar slants% $t forms irregular dry white colonies on nutrient agar% ?henever re:uired a single colony of the culture is inoculated into nutrient broth of 7@ ml in 988 ml conical flas' and the growth condition are maintained at ;A8 C temperature and placed in 97@ rpm orbital sha'er% The medium composition re:uired for growth of culture will be Urea buffer and Calcium chloride% "o study the Me(hani(a) properties of (on(rete The investigation will be carried out to study the echanical properties of concrete% Concrete

cubes of 988 mm & 988 mm & 988 mm will be cast and tested to study the compressive strength under a&ial compression for 7Bdays" C8 days" D8 days" 9B8 days" 7A8 days and ;C@ days% cylinders will be cast and tested to study the split tensile strength% for 7Bdays" C8 days" D8 days and 9B8 days "o study the stress3stain 2ehavior of (on(rete The investigation will be carried out to study the stress!stain behavior of concrete% Cylinders will be cast and tested for 7B days" C8 days" D8 days and 9B8 days% "o study the f)e4ura) 2ehavior of (on(rete The investigation will be carried to study the fle&ural behavior of concrete% simply supported beams of 988mm & 9@8mm & 9788mm consisting of balanced section will be cast and tested% The beams will

be cast treating with and without bacteria% 3rom the developed stress!strain curves" an analytical model will be predicted to develop the theoretical moment!curvatures% These theoretical values of moment!curvatures will be compared with the observed e&perimental values for validation purpose% "o study the strength )oss$ 1eight )oss$ A(id Dura2i)ity 5a(tor and A(id Atta(- 5a(tor of (on(rete Cubes of si,e 988mm & 988mm & 988mm will be cast and cured% +fter 7B days of curing in water cubes will be immersed in @E concentrated H 724F and another sets of cubes are immersed in @E concentrated HCG% The percentage weight loss" percentage compressive strength loss will be ta'en for a set of cubes at 7B days" ;8 days" F@ days" C8 days" A@ days" D8 days and 98@ days% +fter 7B days of casting" each cube will be tested for weight and lengths% +n accelerated e&perimental test program is conducted on ordinary )ortland cement concrete% 2pecimens will be subjected to @E solutions of both sulfuric acid and hydrochloric acids individually% Cubes will be continuously immersed in solution% The specimens will be arranged in the plastic tubs in such a way that the clearance around and above the specimen is not less than ;8 mm% The solution will be changed for an interval of every 9@ days after ta'ing the measurements% The response of the specimens to the solutions is evaluated through change in appearance" weight" compressive strength" thic'ness and solid diagonals% Two specimens from each group will be used for testing after every 9@ days of immersion% /efore testing" each specimen will be removed from the baths" and brushed with a soft nylon brush and rinsed in tap water% This process removes loose surface material from the specimens% 3or determining the resistance of concrete specimens to aggressive environment such as acid attac'" the durability factors are proposed by the author" with the philosophy of +2T 9DDA" as the basis% +2T C CCC -

C CCC - 9DDA considers the standard test method for resistance of concrete

to rapid free,ing and thawing and the durability factors will be defined in terms of relative dynamic modulus of elasticity% $n the present investigation" $ will derive the 0+cid Durability 3actors1 directly in terms of relative strengths% The relative strengths are always with respect to the 7B days value # i%e% at the start of the test *% The e&tent of deterioration at each corner of the struc' face and the opposite face is measured in terms of the solid diagonals #in mm* for each of two cubes and the 0+cid +ttac' 3actor1 #++3* per face will be calculated as follows% ++3 H #Goss in mm on eight corners of each of 7 cubes* I F

The weight loss and strength loss as a percentage of the ;8" F@" C8" A@" D8 and 98@ days strength will be been calculated and tabulated% "o study the a((e)erated (orrosion of (ra(-ing in (on(rete 4ne of the most significant causes of deterioration of reinforced concrete structure is associated with the corrosion of reinforcement% Corrosion of reinforcing steel in concrete and resultant crac'ing and spalling of concrete is an important technological problem% 3or the reinforcing bars to get corroded to appreciable and measurable e&tent" it could ta'e a long time if the specimens are simply e&posed in marine environment% However there are several methods to study the phenomenon in the laboratory under accelerated corrosion conditions% /asically there are two methods for testing the accelerated corrosion of steel in concrete% 4ne involves passing a constant current through reinforcement and other involves 'eeping the voltage constant throughout the test% This study presents the results of modified method of constant voltage techni:ue suitable for accelerated crac'ing studies in the beams having the reinforcement of 98 mm diameter HJ2D bar at different effective covers ranging from 98 mm to F8 mm at all four corners of each beam% /eams of 9@8 & 9@8 & @88 mm will be cast and cured for 7B days in fresh water%

S'MMAR+ Thus $ hypothesi,e that the addition of Bacillus subtilis bacteria improves the compressive strength of cement mortar and at a particular microbial cell concentration i%e% 98 @Iml compressive strength of cement mortar is ma&imum%

The addition of Bacillus subtilis bacteria improves the hydrated structure of cement mortar% The compressive strength of cement mortar is ma&imum with the addition of Bacillus subtilis bacteria for a cell concentration of 9&98@ cells per ml of mi&ing water%

$n ordinary grade concrete the compressive strength is increased up to 9;%D;E at 7B days by addition of Bacillus subtilis bacteria when compared to conventional concrete% $n standard grade concrete the compressive strength is increased up to 9F%D7E at 7B days by addition of Bacillus subtilis bacteria when compared to conventional concrete%

$n ordinary grade concrete the split tensile strength is increased up to 97%C8E at 7B days by addition of Bacillus subtilis bacteria when compared to conventional concrete%$n standard grade concrete the split tensile strength is increased up to 97%8DE at 7B days by addition of Bacillus subtilis bacteria when compared to conventional concrete% 2tress!strain behaviour of Conventional K /acterial concrete is observed% The /acterial concrete mi&es have shown improved stress values for the same strain levels compared to that of Conventional concrete mi&es% +verage values of strain at pea' stress for Conventional K /acterial concrete are very close to the value of strain at pea' stress for conventional concrete in a&ial

compression which is 8%887 as per $2 F@C!7888% Deflections for conventional and bacterial concrete beams at service loads are less than the ma&imum permissible deflection of F%B mm i.e 2pan I 7@8 specified by $2 F@C!7888% Thus the use of conventional and bacterial concrete did not violate the serviceability norms of the codes of practice% Goad deflection behavior of conventional and bacterial concrete beams is observed to be similar e&cept the increased values of loads at ultimate and at first crac'% Theoretical moment!curvature relationships for conventional and bacterial concrete beams followed similar pattern as that of e&perimental values% 3rom the durability studies" in ordinary grade and standard concrete the percentage weight loss and percentage strength loss of cubes immersed in @E H 724F and @E HCG indicates that /acterial concrete has less weight and strength losses than the conventional concrete% Durability studies carried out in the investigation in ordinary grade and standard concrete through acid attac' test with cubes immersed in @E H 724F and @E HCG indicates that bacterial concrete is more durable in terms of 0+cid Durability 3actors1 than conventional concrete and also bacterial concrete is less attac'ed in terms of 0+cid +ttac' 3actors1 than conventional concrete% The 0Charge Deterioration 3actors1 in ordinary grade and standard concrete for a given effective cover" at a given time for bacterial concrete are lowest than that of conventional concrete% $t is concluded that that bacterial concrete will have the higher life compared to conventional concrete% +ll the above investigations will be carried out on higher grades along with the investigations on crac' healing patterns in free,e -thaw cycles environment and the effect of bacteria in concrete subjected to high temperatures will also be investigated along with cost effectiveness of bacterial concrete when compared to other self healing methods of concrete%

RE5ERENCES 9% /ang" 2%2%" >alinat" J%6% and .ama'rishnan"5% 0Calcite precipitation induced by polyurethaneimmobili,ed /acillus pasteurii1 Enzyme and Microbial Technology 5ol%7B #7889* pp=F8F-F8D 7% Chiara /arabesi" +lessandro >ali,,i" >iorgio /runella )erito 0/acillus subtilis >ene astromei" Cluster ila .ossi" Elena" Tamburini and in Calcium Carbonate

$nvolved

/iominerali,ation1 Journal o Bacteriology" Jan%788A" pp%77B! 7;@ ;% .ami'rishnan" 5%" )anchalan" .%6% and /ang" 2%2% 0$mprovement of concrete durability by bacterial mineral precipitation1 Proceedings !"# 99" Torino" $taly" 788@% F% 5%.ama'rishnan" 6%)%.amesh and 2%2%/ang" 0/acterial Concrete1 Proceedings o SP!E 5ol%F7;F p%9CB!9AC" 2mart @% ?illem De aterials%

uync'" 6athelijn Co&" Nele De /elie" ?illy 5erstraete%" /acterial carbonate

precipitation as an alternative surface treatment for concrete% Elsevier Journal" Construction and /uilding C% /ang" 2 2 and .ama'rishnan 5 0 icrobiologically!Enhanced Crac' .emediation# EC.*1 )roceedings of the $nternational symposium on $ndustrial +pplication of Taegu" 6orea" 7889% A% /achmeier 6" ?illiams + E" ?arminton J and /ang" 2%2% 0Urease activity in - induced calcite precipitation1 Journal o Biotechnology, D;= 9A9!9B9" 7887 B% /ang" 2%2% and .ama'rishnan" 5%" L icrobiologicallyMEnhanced Crac' .emediation # EC.*"L )roceedings of the ocrobiological 2ociety of 6orea" 2eoul" 6orea" 7889% icrobiologically icrobial >enomes" aterials 77 #788B* pp%BA@!BB@%

D% /ang" 2%2% and .ama'rishnan" 5%" L/acteria for .epair of Crac's in Concrete"L the Transportation .esearch /oard" ?ashington" DC" 7888

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Structural Engineering, &ol. !, 'o. (, pp. )*-*+, ,ctober (--.. 97% H%2% )atil" .aijiwala D%/%" Hingwe )rashant and /habhor 5ijay" 0/acterial Concrete ! + 2elf Healing Concrete1" $nternational Journal of +pplied Engineering .esearch" 5olume ;" Number 97 #788B*" pp% 9A9D-9A7@% 9;% .amachandran" 2%6%" .ama'rishnan" 5%" and /ang" 2%2%" L.emediation of Concrete Using icroorganisms"L +merican Concrete $nstitute aterials Journal" DB=;!D" 7889 ulu'utla 2%" )erformance of /acterial icroscopy" 2an

9F% .ama'rishnan 5" /ang" 2 2" )anchalan . 6"

Concrete% )roceedings of the 7Fth $nternational Conference on Cement Diege" C+" 7887

9@% .ami'rishnan" 5%" )anchalan" .%6% and /ang" 2%2% 0$mprovement of concrete durability by bacterial mineral precipitation1 )roceedings $C3 99" Torino" $taly" 788@% 9C% .ama'rishnan" 5%" /ang" 2%2%" and )anchalan" .%6%" L/acterial Concrete - + 2elf!.epairing /iomaterial"L )roceedings of the 98th $nternational Congress on )olymers in Concrete and $C)$I$C.$ $nternational Concrete .epair ?or'shop" Honolulu" H$" 7889% 9A% .ama'rishnan" 5%" .amesh" 6%)%" and /ang" 2%2%" L/acterial Concrete"L )roceedings of the 2)$E 7888 2ymposium on 2mart 9@ 7888 9B% 2anthosh 6 ." .ama'rishnan 5" Du'e E 3 and 2 2 /ang 02E $nvestigation of icrobial Calcite )recipitation in Cement1 )roceedings of the 77 nd icroscopy" pp%7D; - ;8@" ontreal" Canada" 7888% aterials and E 2" elbourne" +ustralia" December 9;!

$nternational Conference on Cement

9D% 2toc's!3ischer" 2%" >alinat" J% 6%" and /ang" 2%2%" 0 icrobiological precipitation of CaC4;1" Soil Biology and Biochemistry" 5ol% ;9" 9DDD" pp% 9@C;!9@A9 78% 5%.ama'rishnan" 6%)%.amesh and 2%2%/ang" 0/acterial Concrete1 )roceedings of 2)$E 5ol%F7;F p%9CB!9AC" 2mart aterials%

79% ?illem De

uync'" Dieter Debrouwer" Nele De /elie" ?illy 5erstraete 0/acterial carbonate

precipitation improves the durability of cementitious materials1 Cement and Concrete .esearch" 5ol%;B #788B*" pp%988@-989F% 77% 5%2warnalatha" 6%)%.amesh" 5%.ama'rishnan and 2%2%/ang%" E&perimental $nvestigation on the plastic shrin'age crac' reduction potential of /acteria in concrete% Proceedings o !nternational con erence on ad%ances in concrete and construction, !"/""-0ec.-(--1 " Hyderabad" pp%97@!9;F% 7;% Nhong" G%" +nd $slam" %.%" 0+ New icrobial )rocess and its $mpact on 3racture

.emediation1" A8th +nnual Technical Conference and E&hibition of the 2ociety of )etroleum Engineers" Dallas" Te&as" 4ct 77!7@" 9DD@%