Microbiology applications in civil engineering

1. Microbial Biotechnology Applications
in Civil and Environmental Engineering
Melvin Joe M
Department of Agricultural Microbiology
College of Agricultural Sciences, SRM Institute
of Science and Technology
melvinjm@srmist.edu.in

2. What is microbial
biotechnology
Importance of
microbial in the context
of civil and
environmental
engineering
Microbial
biotechnology in
constructional
technology
Bio-cement, bio-bricks,
and bio-concrete
Concrete microbial
microbial diversity and
their significance in civil
engineering
Soil microbiology and
their importance in
construction
engineering
Aero-microbiology and
their significance in
indoor air quality and
bio-deterioration of
buildings
Environmental
pollution and role of
microbial
biotechnology in the
remediation
Conclusions and the
future

3. NATURE OF BIOTECHNOLOGY AND
INDUSTRIAL MICROBIOLOGY
“any technological application
that uses biological systems,
living organisms, or derivatives
thereof, to make or modify
products or processes for
specific use.”
Microbial biotechnology may be
defined as the study of the large-
scale and profit-motivated
production of microorganisms or
their products for direct use, or
as inputs in the manufacture of
other goods.

4. CHARACTERISTICS
OF MICROBIAL
BIOTECHNOLOGY
• The discipline of microbial
biotechnology is often divided
into sub-disciplines such as
medical microbiology,
environmental microbiology, food
microbiology and industrial
microbiology.
• In Microbial Biotechnology the
immediate motivation is profit
and the generation of wealth.

5. Multi-disciplinary or Team-work Nature of
Industrial Microbiology
In industrial microbiology the
microorganisms involved or their
products are very valuable.
Scale is large and the organisms
may be cultivated in fermentors as
large as 50,000 liters or larger.
Chemical or production engineers,
biochemists, economists, lawyers,
marketing experts, and other high-
level functionaries.

6. Microbiologist or
biotechnologist -key role
in his organization.
• Selection of the organism to be used in the
processes;
• Choice of the medium of growth of the organism;
• Determination of the environmental conditions for the
organism’s optimum productivity i.e., pH,
temperature, aeration, etc.
• monitor the process for the absence of contaminants,
and participate in quality control to ensure uniformity
of quality in the products;
• Proper custody of the organisms usually in a culture
collection, so that their desirable properties are
retained;
• improvement of the performance of the
microorganisms by genetic manipulation or by
medium reconstitution.

7. Microbiology in Civil
Engineering
• Public health and environmental engineers
are aware of the importance of microbial
activity,
• many civil engineers do not appreciate the
part microbiological process
• biodeterioration of concrete and other
construction materials, alteration of soil
and rock properties, clogging of boreholes,
distribution and irrigation systems, and
biofouling in embankment dams.
• There is a need for greater interaction
between microbiologists and engineers in
this respect.
• Recent advances in applied microbiology
and biochemistry could usefully be
extrapolated to fields of civil engineering

8. Biological process of soil
improvement in civil
engineering
• Bio-mediated soil improvement technique
greater potential in geotechnical
engineering applications
• 109–1012 organisms per kilogram of a soil
mass close to the ground surface.
• calcite precipitate to modify some
engineering properties of the soil-
microbially induced calcite precipitation
(MICP), to precipitate calcium carbonate
into the soil matrix.
• calcium carbonate produced binds the soil
particles together (thereby cementing and
clogging the soils), and hence improves
the strength and reduces the hydraulic
conductivity of the soils.
• MICP can be a practicable alternative for
improving soil-supporting both new and
existing structures

9. Current soil improvement practices and their
alternatives
• A common approach is to inject synthetic
man-made materials, such as micro-
fine cement, epoxy, acrylmide,
phenoplasts, silicates, and
polyurethane into the pore space to
bind soil particles together.
• accomplished a variety of chemical,
jetting, permeation grouting techniques.
• These approaches create
environmental concerns and are
increasingly under the scrutiny of public
policy : all chemical grouts except
sodium silicate are toxic and/or
hazardous
• Bio-augmentation -exogenous bacteria cultured in the
laboratory are added into the soil
• biostimulation approach modifies the local environment
by injecting nutrient media

10. Urease-aided
calcium
carbonate
mineralization
• The process of urease-aided calcium carbonate
mineralization is triggered by the catalytic action of urease
in the hydrolysis of urea
• The products of the reaction are carbonic acid and
ammonia:
H2N-CO-NH2+2H2O→ureaseH2CO3+2NH3
• The products equilibrate in water to give bicarbonate,
ammonium and hydroxide ions, respectively:
H2CO3⇄HCO3-+H+(5)NH3+H2O⇄NH4
++OH-
• The production of hydroxide ions from reaction brings
about an increase in pH, which in turn leads to the
formation of carbonate ions
HCO3-+OH-⇄CO3
2-+H2O
• In the presence of dissolved Ca2+, the ions combine and
calcium carbonate precipitates:
Ca2++CO3
2-→CaCO3(s)
The overall process can thus be presented:
H2N-CO-NH2+2H2O+Ca2+→urease2NH4
+ + CaCO3(s)

11. Urease-producing
microorganisms
• Urease enzyme producing bacteria and are
used in bio-mediated soil improvement
technique
• Urease-producing microorganisms can be
divided into two different classes based on their
response to high presence of ammonium.
• The first group includes the bacteria whose
urease activity is not repressed due to high
ammonium concentration (Table 1).
• While the second group includes Bacillus
megaterium, Alcaligenes eutrophus, Klebsiella
aerogenes and Pseudomonas aeruginosa ,
whose urease activity is repressed by high
ammonium concentrations.
Fig. 1. Microbial calcite precipitation process by urea
hydrolysis (DeJong et al., 2010).

12. Fig. 2. Comparison of soil particles sizes, geometric
limitations and microorganisms DeJong et al. (2010).
Soil particles sizes, geometric limitations and microorganisms
Microbes are viable to move freely within
the voids of the soil aggregates, their
movements are restricted by the narrow
pore sizes formed by fine-grained soils.
Bacteria sizes range between 0.5 μm and 3
μm, as such they are not likely to pass
through pore spaces smaller than 0.4 μm.
Likewise, fungi and protozoa require pore
sizes greater than 6 μm to pass

13. • Biotechnology for the production of
construction materials– low cost due to use
of mining or organic wastes as raw
materials;
• lower cost in comparison with the products
of chemical industry
• Simpler and less energy consuming
technology
• Lower toxicity of biomaterials than
chemical materials
• Sustainability of the biotechnological
production.
• Geotechnical engineering for bio-
aggregation, bio-clogging, and bio-
cementation of porous soil or fractured
rocks in situ
• Low viscosity of bio-grouting and bio-
cementing solutions and deep penetration
into porous soil or fractured rocks;
• Control rate of biochemical reactions in situ
by the concentration or activity of biomass
or enzyme
• Ability for self-multiplication (proliferation) of
microbial cells in situ
• Better public acceptance of biotreatment
than chemical treatment

14. • Biocement stimulates native soil bacteria to
connect soil particles through a process known
as microbially induced calcite precipitation
• Strong and renewable building material with
minimal impact on the environment.
• Compared to the production process of
traditional cement, biocement uses less energy
and generates less CO2 emissions.
• Combination of biomass, aggregate,
renewable nutrients and minerals that are
placed into moulds and then treated with a type
of bacteria (Sporosarcina pasteurii) that is fed
with calcium ions and water.
• This results in the production of a calcium
carbonate shell that can be used to create a
'natural' bio-cement brick.
• The process takes less than three days and
is said to simulate the actions used by corals.
Bio-cement

15. Nanotechnology in
Cementitious Materials
• FIGURE: (A) TEM image of clusters of C-S-H, the
inset is a TEM image of tobermorite (B) the molecular
model of C-S-H: the blue, white spheres are oxygen
and hydrogen atoms of water molecules, respectively
• Biological synthesis of NPs also eliminates high energy-
consuming processes
• Variety of nanoparticles (e.g., SiO2, ZrO2, TiO2, CuO), the
setting time and diffusivity of concrete are reduced, while the
strength and high-temperature stability are increased
• Among multiple phases of hydration product that are formed
during Portland cement hydration, calcium silicate hydrate (C-
S-H) is the main phase in hydrated Portland cements.
• Additionally, the presence of the NPs at the interface between
aggregates and cement matrix promotes localized nucleation
of hydration products- ITZ is usually the weakest zone in
concrete.

16. Bioconcrete as a promising
sustainable technology
Surface images of specimens after different
healing time. (A) G Specimens without self-
healing agent; (B) specimens with self-healing
agent
o Concrete is also responsible for about 8% of
global carbon emissions.
o If concrete were a country, it would rank
third in emissions behind China and the
United States.
o Bioconcrete is a promising sustainable
technology which reduces negative
environmental impact caused by
CO2 emissions from the construction sector,
o As well as in terms of economic benefits by
way of promoting a self-healing process of
concrete structures.
o Microbiological and molecular components
are essential to improving the process and
performance of bioconcrete

17. Bacteria based self
healing concrete
• Bio-mineralization techniques
promising results in sealing the
micro-cracks in concrete.
• The freshly composed micro-
cracks can be sealed up by
perpetual hydration process in
concrete.
• The ureolytic bacteria which
include Bacillus Pasteurii,
Bacillus Subtilis which can
engender urea are integrated
along with the calcium source
to seal the freshly composed
micro cracks by CaCO3
precipitation.
• Amelioration of pore structure
in concrete, the bacterial
concentrations were optimized
for better results.

18. Mechanism of
applying the
healing agents in
concrete

19. Compression strength improvement in concrete with
various bacterial strains

20. Concrete microbial community
composition.
• Concrete bacterial community relative
abundance.
• Overall, 50% of sequences were
classified at the phylum level as
Proteobacteria, 19% as Firmicutes,
14% as Actinobacteria, Of the
Proteobacteria,
Gammaproteobacteria were the
most abundant
• Gram-positive taxa account for
∼33% of all reads. Almost 19% of
sequences were classified as
Firmicutes, which can form spores,
potentially allowing them to survive
in dormant states in the harsh
concrete environment.

21. Air-borne
transmission
Transmission of microbial aerosols to a
suitable port of entry, usually the
respiratory tract
Microbial aerosols -suspensions of dust or
droplet nuclei made up wholly or in part
by microorganisms -- may be suspended
and infective for long periods of time
Examples of air-borne diseases include
tuberculosis, influenza, histoplasmosis,
and legionellosis

22. Indoor microbiology:
Environmental factors and
subsequent effects
• a Sunlight (both UV light and visible
light survival of microorganisms living in
the built environment.
• b Biofilms sinks and showers in
bathrooms
• c Moisture from daily activities,
biofilms while and high relative humidity
increases the rate of aerosolized
microbial cells and spores.
• d Frequented household items, such
as chairs, plentiful supply of nutrients
• e Microenvironments within carpet
can create pockets of high relative
humidity that can aid in the growth,
prolonged survival, and transfer of
microorganisms from fomite to
individual.
• f Ventilation through windows
provides air exchange that aids in the
reduction of potentially contaminated air.
• g Humans, insects, pets, and other
occupants exchange microorganisms

23. Indoor
microbiology:
Microorganis
ms in indoor
environment
• Results displayed in Figure demonstrate that the PM10 and PM2.5 fractions of resuspended
floor dust are enriched with bacteria, compared to indoor air, ventilation duct supply air, and
outdoor air.
• Samples show heavy representation from the dominant bacteria previously found to be
associated with human skin, hair, and nostrils — Proprionibacterineae, Staphylococcus,
Streptococcus, Enterobacteriaceae, and Corynebacterineae — comprise 17%, 20%, and
17.5% of all bacteria in samples of indoor air, floor dust, and ventilation duct supply air,
respectively

24. Health Effects and Indoor Air Quality
Health Issues
■ Nasal congestion and runny nose
■ Watery, burning eyes
■ Sore throat and hoarseness
■ Dry, irritant-type cough
■ Tight chest, burning sensation, wheezing,
shortness of breath
■ Nosebleeds, coughing blood (rare)
■ Skin and mucous membrane irritation, rashes
■ Exhaustion, severe fatigue
■ Memory and cognitive problems
■ Gastrointestinal problems such as nausea,
vomiting
■ Joint and muscle pain
■ Fever
■ Headaches
Precautions
■ Maintain relative humidity below 60 percent within
buildings;
■ Use an air conditioner or a dehumidifier during humid
months and maintain it properly
■ Provide adequate ventilation in buildings, including
exhaust fans in kitchens and bathrooms
■ Keep bathroom and kitchen surfaces clean
and regularly treat them with disinfecting products
■ Do not place carpeting in bathrooms, basements, or
other areas where humidity is high; and
■ Remove or replace carpets and upholstery if they
cannot be dried out immediately after becoming wet

25. Microbial deterioration and
sustainable conservation of
stone monuments and
buildings
• Geomicrobially induced deterioration of
stone monuments and buildings
contributes to a considerable loss of
world cultural heritage
• The active biodeterioration processes
typically involve biochemical activities
and cooperation among functional
microorganisms in epilithic biofilms,
which assimilate mineral nutrients and
metabolize anthropogenic pollutants
through biogeochemical cycles.
a, b Growth of the fungus Cladosporium sp. on a modern wall painting
White Carrara marble -black discolorations by fungi and lichens

26. Influence of Environment on Microbial Colonization of Historic Stone Buildings
Trentepohlia algae forming deep red discoloration Black cyanobacterial crusts on the walls of the Our Lady
of the Immaculate Conception cathedral

27. • increase of awareness due to community
involvement in enhancing moisture control
• improvement of cleaning processes and the use
of air conditioning systems,
• regular inspection and maintenance regimes for
buildings and
• cleaning of heating and air conditioning units and
associated replacements of filters.
• More emphasis must be focused on simple
prevention measures such as the cleaning of
dust layers and frequent observation of objects.
• Biocide treatments must be applied with extreme
caution
• More effort is necessary in the development of
alternative decontamination methods, e.g., the
gamma radiation modification of light and micro-
climates

28. Biosurfactants
• Chemically, BS is a complex molecules consisting of
lipopeptides,
glycolipids, polysaccharide protein complex, fatty acids
and phospholipids
• Bio surfactants (BS) reduce surface (ST) and interfacial
tensions
between individual molecules
• Biodegradability, low toxicity, and renewable nature
• Ability to withstand high temperature and tolerate high
salt concentration

29. Types of biosurfactants produced by various
Microorganisms
Biosurfactant class Microorganisms Reference
Glycolipids
Rhamnolipids Pseudomonas aeruginosa Lang and Wullbrandt (1999)
Trehalose lipids Rhodococcus erythropolis, Lang and Philip (1998)
Sophorolipids Torulopsis bombicola, Gobbert et al. (1984)
Lipopeptides
Surfactin/Iturin/ Bacillus subtilis Arima et al. (1968)
Lichenysin Bacillus licheniformis Yakimov et al. (1995)
Fatty acids, neutral lipids and phospholipids
Fatty acids Corynebaterium lepus Cooper et al. (1978)
Neutral lipids Nocardia erythropolis MacDonald et al. (1981)
Phospholipids Thiobacillus thiooxidans Beeba and Umbreit (1971)
Polymeric Biosurfactants
Emulsan/Biodispersan Acinetobacter calcoaceticus Rosenberg et al. (1988)
Alasan Acinetobacter radioresistens Navon-Venezia et al. (1995)
Liposan Candida lipolytica Cirigliano and Carman (1984)

30. Biochemistry of
Rhamnolipid
biosynthesis
• Parallel linked RhlA and
PhaG metabolic pathways
via fatty acid de novo
biosynthesis leading to
synthesis of rhamnolipids
and PHA (Choi et al. 2013)

31. Genetics of
rhamnolipid
synthesis
Model for rhamnolipid production and regulation in P. aeruginosa (Lang and Wallbrant, 1999)

32. Enhanced production of biosurfactant / production by mutant
and recombinant strain
Microorganism Methods of strain improvement Status of improved strain References
Bacillus subtilis ATTC
21332
UV mutation 3 times more than parent
strain
Mulligan et al. (1989)
Recombinant
B. subtilis M113
Cloning of plasmid PC112
containing lpa-14 gene.
8 times enhanced
production
Ohno et al. 1995
Pseudomonas aeuroginosa
PTCC 1637
Random mutagenesis with
nitrosoguanidine
10 times more than parent
strain
Abbas Tahzibi et al. (2003)
Recombinant
B. subtilis ATCC 21
Genetic engineered peptide
synthetase
Surfactin produced with
less toxicity
Symmank et al. (2003)
Recombinant E.coli strain Expression of only rhlAB gene in E.
coli.
Ability to produce RL Wang et al. (2007)
Recombinant P.putida strain Cloning of rhlAB genes and rhlRI
quorum sensing system
Higher production of RL Cha et al. (2008)
Recombinant E.coli strain Cloning of genes sfpO, srfA of B.
subtilis SK320
Two fold increased
production
Sekhon et al. (2011)

33. Application of different types of biosurfactants in
environmental remediation
Type of Biosurfuctant M.O’s Involved Usefulness as a Biocontrol agent References
Rhamnolipid Commercial
Biosurfactant
micellar-enhanced filteration of heavy metals Cu,
Zn, Ni, Pb, Cd,
El-zeftawy and Mulligan
(2011)
Rhamnolipid Commercial
Biosurfactant
Addition of 1 -0.5% rhamnolipid for copper
removal
Dahrazama and Mulligan
(2007)
Crude
biosurfactant
Candida lipolytica
UCPO98
Crude biosurfactant removed 96% Zn and Cu and
reduced concentration of Pb, Cd, and Fe from
test specimen
Rufino et al. (2011)
Biosurfactant Fusarium sp. BS8 For microbial enhanced oil recovery (EOR) Quazi et al. (2013)
Biosurfactant Bacillus spp.
SH2O/SH26
oil contaminated vessels/ enhanced
biodegradation of oil sludge
Diab and Din (2013)
Surfactin Bacillus subtilis Sediments contaminated with Zn, Cu, Cd, oil and
grease
Mulligan et al. (1999)
Rhamnolipid Consortium of
bacteria
Positive role in hydrocarbon degradation Chen et al. (2013)

34. Phytomediation and bioaugmentation on
the rhamnolipid mediated
biodegradation of diesel oil
contaminated soils
Control, T1 (Plant alone), T2
(Plant + 2% Rhamnolipid
biosurfactant (BS), T3 (Plant +
bioaugmentation (BA)), T4 (Plant
+ BS + BA), T5 (Plant+ 2%SDS),
T6 (Plant+ 2%SDS + BA), T7
(2%SDS), T8 (2% BS), T9 (BA),
T10 (BS + BA), T11(SDS + BA).

35. Conclusions
• Microbiology of Construction Biotechnology process requires
understanding and strict performance of the biosafety rules aiming to
prevent outbreaks of the infectious diseases during the production of
construction biomaterials or application of microorganisms in
construction process.
• Understanding of microbiology is essential in the development of
biotechnological construction material or biotechnological construction
process but the commercial biomaterial or bioprocess must be made.
• Geotechnical or environmental applications of microbial processes in
the field may require to complexity of the factors or either partnership
with microbiologist or understanding of the basic principles of
microbiology.
• The characterization of the physicochemical interactionsbetween
substrates and microorganisms and the adhesive properties of the
microorganisms themselves to be studied for better preservation of civil
structures