Biotechnology for Environment Protection

8/14/2019 Biotechnology for Environment Protection

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BIOTECHNOLOGY FORENVIRONMENT PROTECTION

Irfan D. Prijambada

Fac. of Agriculture,Gadjah Mada University


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THE BEGINNINGS

World War II brings an unprecedentedgrowth in the economy and business Breakthroughs in organic chemistry

War effort and postwar economic boom Huge volumes of wastes generated by this

evolving industry

Lack of knowledge on environmental andhealth ramifications of waste disposal


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LANDFILLS

Majority of waste is deposited inlandfills

First sanitary landfills developed in

1920sSolid wastes are spread out in thin

layers, compacted, and covered dailywith fresh clay or plasticSolved problem of foul smell and reduced

incineration needs

But by 1960s, evident that not capable ofcontaining groundwater contamination


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LANDFILLS

Modern landfills are lined with clay andplastic before being filled with garbage

Bottom is covered with a second

impermeable liner, usually made ofseveral layers of clay, thick plastic, andsand This liner collects leachate, rainwater

contaminated as it percolates through thesolid waste, which is then pumped frombottom of the landfill, stored in tanks, andsent to a sewage treatment plant


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LANDFILLS

Anaerobic conditions are createdwithin landfill wasteSlow stabilization of waste mass occurs,

producing methaneExplosive and toxic over long periods of

time One study found that aerobic degradation of

waste within a landfill can significantly

increase the rate of waste decompositionand settlement, decrease production ofmethane gas, reduce level of toxic organicsin leachate, and decrease amount of

leachate that need treatment


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RESULTS OF MISMANAGEMENTOF WASTE

Rachel Carson publishes SilentSpring in 1962 Provoked widespread public alarm with

her attack on pesticide usage,emphasizing the unintended ecologicalconsequences of pesticide use

Illustrated interconnected web of lifeand how such elixirs of death werestored in humans


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CONTAMINATION INCIDENTS

Accident in Japan CONTAMINATION INCIDENTS IN JAPAN

Accident in USA CONTAMINATION INCIDENTS IN USA


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CONTAMINATION INCIDENTS INJAPAN

Two cases in Japan make worldwideheadlinesHundreds paralyzed due to mercury

poisoning caused by eating shellfishaffected by products of a chemicalplant

Rash of miscarriages blamed on use of

rice-cooking oil contaminated with PolyChlorinated Biphenyls (PCBs)

CONTAMINATION INCIDENTS


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CONTAMINATION INCIDENTS INUSA

Love Canal, August 1977

Black sludges bleed through basement walls in

suburban subdivision of Niagara Falls, NY

Reports of benzene fumes in kitchens,

headaches, skin problems, respiratory

discomfort

Shortly, dioxin detection, miscarriages and birth

defects

Government pays for evacuation, at cost of $30

million


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Evacuation process piecemeal over three years

amidst climate of high tension, misinformation,

broken promises

May 19, 1980 Love Canal activists take two

government representatives hostage overnight

Caused by one hundred thousand drums of

chemical waste dumped into an abandoned canal

by Hooker Chemical and Plastics Corporation

Shows that some kind of regulation is needed

CONTAMINATION INCIDENTS INUSA


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MOST COMMON CONTAMINANTS

Commercial Hydrocarbons

gasoline

diesel and jet fuel naptha: raw material used in industry

domestic heating oil

Chemicals called BTEXcompounds

Benzene, Toulene, Ethylene,Xylene


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MOST COMMON CONTAMINANTS 2

Organo-halogenated compounds(solvents) trichloroethylene, tetrachloroethane, etc

Heavy hydrocarbons crude oil: pipeline, tanker and rail spills

heavy fuels from electric plants

tars

creosotes used in wood treatments


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MOST COMMON CONTAMINANTS 3

Heavy metals

Explosives


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CHEMICALS WHICH AREDIFFICULT TO DECOMPOSE

Trichloroethylene (TCE) - threatenswater supplies

Perchloroethylene (PCE) - a dry-cleaning solvent

PCBs and Dioxin

Arsenic, chromium, and selenium(these have been stabilized bybacteria in the laboratory)

DDT


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POLLUTED SITES

Accidental spills

Service stations

Old Air Force bases Storage tanks and pipelines

Chemical plants and otherindustrial sites

Unauthorized dump sites


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BIOREMEDIATION

Bioremediation is the use of livingmicroorganisms to degradeenvironmental contaminants in the soil

and groundwater into less toxic, ornontoxic materials.

These microorganisms can be indigenous,commercial bacterial mixtures (bag ofbugs or bug n a bag) or may begenetically engineered.


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BIOREMEDIATION 2

Bacteria feed on organic waste andderive nutrition for growth andreproduction. This is familiar to all as

the decay of dead animals andvegetable matter.

Municipal wastewater treatment plants

have been using this technology fordecades. Bioremediation is anapplication of the same principles in a

different setting.


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BIOREMEDIATION 3

Over time, Mother Nature usuallyheals herself. Adding large amountsof certain enzymes and bacteria

hastens the decay. Utilizingbioremediation speeds up theprocess by increasing the rate of

bacterial metabolism and growth.


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USES OF BIOREMEDIATION

Bioremediation can be used todecompose or degrade:

Crude oil spills

Sewage effluent

Chlorinated and non-chlorinatedsolvents in the industrial areas


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USES OF BIOREMEDIATION 2

Coal Products: phenols andcyanide

BTEX compoundsAgricultural chemicals and

pesticides in groundwater and

rivers


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USES OF BIOREMEDIATION 3

Gasoline and fuel oilcontamination

Creosote contaminants(wood preservatives)

Ethylene glycol (antifreeze),

methanol, methylethylketone(MEK), ethers


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REASONS TO USEBIOREMEDIATION

Bioremediation can be cost effectivebecause:

Contamination can often be treatedin place, minimizing sitedisturbance.

Natural microbial processes can beused at some sites.


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EFFECTIVENESS

Biodegradation is not veryeffective at sites with highconcentrations of the following

materials which are toxic tomicroorganisms

Metals-solidification/stabilization

is the usual treatment processHighly chlorinated organics

Inorganic salts


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DISPOSING OF HEAVY METALS

Heavy metals are notbiodegradable, but bacteria canbe used to concentrate them into

a more easily disposable form.Uranium: iron-eating bacteria

can remove low levels of

radioactive waste from water.Mercury: experiments with

bacteria are on-going.


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MICROORGANISM TYPES

There are large numbers ofmicroorganisms that can use

many of the toxic chemicals as asource of nutrients and energy.Some examples include:

BacteriaYeast

Fungi


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SOME MICROORGANISMS USEDIN BIOREMEDIATION

Microorganism Characteristics Significance

Yeast aerobic/

micro-aerophilic

Degrades complex

compounds

Cyanobacteria aerobic/micro-aerophilic/

anaerobic

Self-sustaining,light is primary

energy source

Oligotrophs aerobic RemovesTRACE

concentrations oforganic substances


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TYPICAL BACTERIA SPECIES INCLUDE:IN DESCENDING ORDER OF OCCURRENCE)

PseudomasArthobacter

Alcaligenes

Corynbacterium Flavobacterium

Achrombacter

Acinetobacter Micrococcus

Nocardia

Mycobacterium


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EXAMPLES OF MICROBES USED FORSPECIFIC CHEMICALS

Compound Name Microorganisms Conditions

Aliphatics

(non-halogenated)

Ex. Acrylonitrile

Mixed culture and

activated sludge

Aerobic

Aliphatics(halogenated)

Ex.Trichloroethane

Marine bacteria,sewage sludge,

soil bacteria,methanogens

Aerobic +Anaerobic

Aromatic

compoundsEx. BTEX,

creosol, phenol

Pseudomonas spp.,

Bacillus spp.,Rhodococcus spp.,

Mycobacterium

spp.

Aerobic +

Anaerobic


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BENEFICIAL CHARACTERISTICS

Beneficial characteristics ofbacteria for bioremediation mustinclude the following:Consume organic waste

Grow and reproduce rapidly in selectedenvironment

Digest the waste quickly andcompletely


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BENEFICIAL CHARACTERISTICS 2

Work without causing odors orpoisonous compounds

Non-pathogenic - (Does not causedisease in humans or animals)


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CLASSES OF BIOREMEDIATION2

Anaerobic (without oxygen) -Microorganisms break down

chemical compounds to release theenergy required to function. Aselectron acceptors, they utilize:

nitratessulfates

carbon dioxide

ferrous metals (such as iron)


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HOW BIOREMEDIATIONWORKS...

Many naturally occurringmicroorganisms can digest

organic materials such as fuelsor solvents and convert them to: carbon dioxide

water

smaller, less toxic organic compounds


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Basic Metabolism Processof Bacteria

Growth

and

Reproduction

Catalyzed by Enzymes

CELL

ENERGY

SOURCENUTRIENTS

CARBONSOURCE

NEWCELL

MASS

H2O

CO2


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Schematic Diagram ofBiodegradation

Oil

MicrobeCO2+H2O

CO2+H2O

CO2+H2O

Microorganisms eat

oil and other organic

contaminants.

Microorganisms

digest oil and

convert it to CO2

and H 0

Microorganisms

releaseCO2 and

H20

1. 2. 3.


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OPTIMIZATION

To optimize and accelerate thebioremediation of contaminantsfound in water and soil,

selectively adapted microbesare combined with: Food - organic waste containing

water (moisture content between30-80%)

added nutrients (nitrogen,phosphorous, sulfur)


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OPTIMIZATION 2

Oxygen if required (aerobictypes) 3-5 pounds of oxygen perpound of hydrocarbon to be

convertedModerate pH - between 6-9,

neither too acidic nor too alkaline

Moderate Temperatures - 50oto 100o F


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OXYGEN DEMAND VALUES

Oxygen demand values are usedto measure biological treatmentprocesses.

Biological Oxygen Demand(BOD) measures the amount of

oxygen necessary for microbesto remove waste in wastewaterin 5 days at 20oC.


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OXYGEN DEMAND VALUES 2

Chemical Oxygen Demand(COD) measures a chemicals

ability to oxidize toxic chemicalsin 3 hours.

The difference between the two

gives the operating efficiency ofa biological process.


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THE EFFECT OF pH ON THE GROWTH OFSPECIFIC MICROORGANISMS

Microorganism Optimum pH

BACTERIA:

Pseudomonas aeruginosa 6.6 7.0

Bacillus alcolophilus 10.6

Nitrosomas spp. 8.0 8.8

Thiobacillus thiooxidans 2.0 3.5

ALGAE:Cynidium caldarium 2.0

FUNGI:

Physarum polycephalum 5.0


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OPTIMIZATION 3

Enzymes, chemical catalysts tobreak waste materials into smallerpieces

Surfactants (detergents, forexample)

TECHNOLOGY SELECTION


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TECHNOLOGY SELECTIONCRITERIA

The bioremediation technologyfor a site is determined by:

Microorganisms present Site Condition

Quantity and Toxicity of

Contaminants

CATEGORIES OF


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CATEGORIES OFBIOREMEDIATION

Bioremediation treatment applicationsfall into 2 categories:

in situ- soil or groundwater is treated in

the location where found. This is usually themost cost effective method, but can also beslower and hard to manage.

ex situ - requires the excavation of soil orpumping of groundwaterbefore treatment.

EXAMPLES OF IN SITU


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EXAMPLES OF IN SITUBIOREMEDIATION

Bio-venting: air and nutrients arepumped into the soil throughinjection wells to flush out

contaminants. Air Sparging: air or oxygen is

pumped into the groundwater to

flush out contaminants - the airincreases the oxygen concentrationand enhances biological degradation.


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AIR SPARGING


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TYPICAL i it Bi di ti


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TYPICAL in situ BioremediationSystem

Contaminated Zone

OldWaterTable

NewWaterTable

WaterTreatmen

t

Nutrient/

OxygenAddition

Recovery Well

InjectionWell

EXAMPLES OF EX SITU


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EXAMPLES OF EX SITUBIOREMEDIATION

Slurry Phase: a large tank, orbio-reactor contains the soil,water, and added nutrients oroxygen to keep themicroorganisms in the optimum

environment to degradecontaminants.


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Contaminatedliquid

Contaminatedsoil

BIOREACTOR

Liquid outlet

Soil todrying

Temperature

control

AgitatorVapor out

Air inlet

Nutrient

EXAMPLES OF EX SITU


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EXAMPLES OF EX SITUBIOREMEDIATION 2

Solid phase: soil remains on thesite, but is placed in above-groundtreatment areas where moisture,heat, and nutrients or oxygen areadded.


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SOLID PHASE EX SITUBIOREMEDIATION 3

Soil Biopile:

The contaminated soil is piled inlarge heaps and air is pulledthrough with vacuum pumps.


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BIOPILES

Nutrient/moistureGravel

layer

Leachatecollection

Impermeablelayer

Contaminated soil


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SOLID PHASE EX SITUBIOREMEDIATION 3

Composting:

Biodegradable waste is mixed with a

bulking agent such as straw, hay, orcorn cobs, which facilitates the deliveryof water and nutrients.

The three types of composting are:

* Static pile* Mechanically agitated in-vessel

* Windrow composting


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DIAGRAM OF BIOREMEDIATION

Adding OxygenBioventingBiosparging

Adding Oxygenand Nutrients

Biostimulation

AddingOxygen,Nutrients

and Bacteria

Bioaugmentation

Engineered Intrinisic

in situ

Landfarming Bioreactor

ex situ

Bioremediation


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REMEDIATION TIME

in situ bioremediation timedepends on the extent, depth, andconcentration of the

contamination. It varies from 1 - 6years.

ex situ remediation for easilybiodegradable contaminants orwhen bioreactors are used cantake as little as 1-7 months.

CASE STUDIES IN


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CASE STUDIES INBIOREMEDIATION (1)

Van Nuys airport in Southern California

High concentrations of petroleum

hydrocarbons

Bioremediation and bioventing were used

Pollutant levels dropped 80% in 90 days

Site no longer considered a risk

Passive remediation continues until microbes

exhaust their food supply

CASE STUDIES IN


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Minnesota Department of Transportation Biomounds used to clean petroleum wastes

Mounds utilize indigenous bacteria

Petroleum contaminants provide the energy Manure provides the nutrients

Wood chips allow the entry of oxygen

Plastic sheeting provides warmer

temperatures for bacteria growth

CASE STUDIES IN


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Alaska

Bioventing used to clean diesel fuels at

Shemya Air Force station

Landfarming succesfully demonstrated

at Fairbanks to treat soil at the airport,

and is now being used at other sites in

the state

CASE STUDIES IN


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Hilo, Hawaii

Biosparging and land treatment

used to degrade hydrocarbon

contaminated soils and

groundwater.

CASE STUDIES IN


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Fort Polk, Louisiana

Landfarming used continuously for over

10 years to treat petroleum spills.

Plants sensitive to petroleum wastes

are used to give indication of

completeness of the process.

Cleaned soil is removed and more

contaminated soil is added.


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ISSUES FOR DISCUSSION (2)

Not all bacteria or microbes aregood.

Some remediation studies have

shown that there is spontaneousmutation in some bacterialpopulations after remediation

efforts. Could this be bad forhumans, animals, and plants?


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ISSUES FOR DISCUSSION (3)

Some companies are usinggenetically alteredmicroorganisms. This leads to a

discussion of genetics, naturalselection, and ethics.

Is it OK to bring in exogenous

microbes? What if they dont dieafter the contaminat is degraded?


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METHODS FOR ANALYSIS OF DNA

Size and structure

of individual DNA

molecules

Degree of relatedness

between molecules by

hybridization procedures

Plasmid DNA

Chromosomal DNA

Drawbacks:i. Lack of stability in some strains

ii. Strains not containing plasmids

iii. plasmid transfer among strains

PCR-based techniques

Gene sequencing Profiling from electrophoresed

PCR-products

DNA FINGERPRINTING OF ORGANISM


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DNA FINGERPRINTING OF ORGANISM

PCR-based methods

Principle:Based on the specificity of inserted DNA

in transgenic organismsThe inserted DNA are amplified by PCRand then electrophoresed to obteinpatterns which can be mathematicallyanalysed to establish clusters.

Level of resolution at species level,is valuable and reliable forphylogenetical and ecologicalt di

Amplified Ribosomal DNA Restriction Analysis (ARDRA)

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Biotechnology for Environment Protection