The document discusses environmental biotechnology and its applications. It provides details about (1) using microorganisms to treat hazardous wastes and pollution, including bioremediation of contaminated soil and water, (2) the treatment process at a common effluent treatment plant (CETP) that cleans waste water from textile industries, and (3) two case studies on bioremediation of oil-contaminated soil and waste water treatment at a CETP.

2. Environmental Biotechnology
• application of processes for the protection and restoration of the quality of the
environment.
• to detect, prevent and remediate the emission of pollutants into the
environment in a number of ways.
• Solid, liquid and gaseous wastes – modified
– Recycling new products
– Purifying less harmful end products
Chemical materials/processes Biological Technologies
Reduced Environmental Damage

3. Environmental biotechnology can make a significant contribution to sustainable
development.
-Fastest growing
-Most practically useful scientific fields.
Research in genetics, biochemistry and physiology of exploitable microorganisms is
rapidly being translated into commercially available technologies for reversing and
preventing further deterioration of the Earth’s environment.

4. Objectives of Environmental Biotechnology
1. To adopt production processes that make optimal use of natural resources, by
recycling biomass, recovering energy and minimizing waste generation.
2. To promote the use of biotechnological techniques with emphasis on
bioremediation of land and water, waste treatment, soil conservation,
reforestation, afforestation and land rehabilitation.
3. To apply biotechnological processes and their products to protect
environmental integrity with a view to long-term ecological security.

5. Applications of Environmental Biotechnology
• Use of living organisms in hazardous waste treatment and
pollution control. Environmental biotechnology includes a
broad range of applications such as:
– bioremediation
– prevention
– detection and monitoring
– genetic engineering
All for sustainable development and better quality of living

6. Research related environmental biotechnology is vital in developing effective
solutions for mitigating, preventing and reversing environmental damage with the
help of these living forms.
Growing concern about public health and the deteriorating quality of the
environment has prompted the development of a range of new, rapid analytical
devices for the detection of hazardous compounds in air, water and land.
Recombinant DNA technology has provided the possibilities for the prevention of
pollution and holds a promise for a further development of bioremediation.

8. Soil and Land Treatment
As the human population grows, its demand for food from crops increases, making
soil conservation crucial.
Deforestation, over-development, and pollution from man-made chemicals are just
a few of the consequences of human activity and carelessness.
The increasing amounts of fertilizers and other agricultural chemicals applied to
soils and industrial and domestic waste-disposal practices, led to the increasing
concern of soil pollution.
Pollution in soil is caused by persistent toxic compounds, chemicals, salts,
radioactive materials, or disease-causing agents, which have adverse effects on
plant growth and animal health.

10. • During biological treatment soil microorganisms convert organic pollutants to
CO2, water and biomass.
– Aerobic or Anaerobic conditions
– In-situ or Ex-situ (Bioreactors)
• Treatment of soil contaminated with mineral oils
• Solid-phase technologies are used for petroleum-contaminated soils
• Biological degradation of oils has proved commercially viable
• In situ soil bioremediation involve the stimulation of indigenous microbial
populations
natural origin cultivated genetically engineered

11. Waste Water and Industrial Effluents
Water pollution is a serious problem in many countries of the world.
Rapid industrialisation and urbanization have generated large quantities of waste
water that resulted in deterioration of surface water resources and ground water
reserves.
Biological, organic and inorganic pollutants contaminate the water bodies.
In sewage treatment plants microorganisms are used to remove the more common
pollutants from waste water before it is discharged into rivers or the sea.
There is a greater need for processes that remove specific pollutants such as
nitrogen and phosphorus compounds, heavy metals and chlorinated compounds.

12. Five Key Stages in Wastewater Treatment
1. Preliminary treatment – grit, heavy metals and floating debris are removed.
2. Primary treatment – suspended matters are removed.
3. Secondary treatment – bio-oxidize organic materials by activities of aerobic and
anaerobic microorganisms.
4. Tertiary treatment – specific pollutants are removed (ammonia and phosphate).
5. Sludge treatment – solids are removed (final stage).
• Aerobic Biological Treatment
– Trickling filters, rotating biological contactors or contact beds, usually consist of an inert material
(rocks/ ash/ wood/ metal) on which the microorganisms grow in the form of a complex biofilm.
• Activated Sludge Process
– In this process wastewater containing organic matter is aerated in an aeration basin in which micro-
organisms metabolize the suspended and soluble organic matter. Part of organic matter is synthesized
into new cells and part is oxidized to CO2 and water to derive energy.

15. Soil and Land Treatment (Case Study)

16. • It mainly involved biostimulation where organic or inorganic components were
introduced to enhance indigenous microbial growth that directly degrades the
contaminants.
• Biostimulation
– addition of substrates, vitamins, oxygen and other compounds to stimulate microorganism
activity so that they can degrade the petroleum hydrocarbons faster.
– addition of nutrients brought large quantities of carbon sources which tend to result in a
rapid depletion of the available pools of major inorganic nutrients such as N and P.
• Organic waste utilization
– Inadequate mineral nutrient, especially N and P, often limits the growth of hydrocarbon
utilizing bacteria in water and soil.
– Addition of a carbon source as a nutrient in contaminated soil enhances the rate of pollutant
degradation by stimulating the growth of microorganisms.

17. • Organic waste collection
– sewage sludge (SS) and cow dung (CD) were selected as the organic components to be
added individually into the 10% (w/w) used lubricant oil-contaminated soil.
• Microcosm set-up
– 1.5 kg of soil (sieved with 2mm mesh size)
• polluted with 10% (w/w) used lubricating oil (150 g) and left undisturbed for two days.
– 10% of each organic waste namely CD and SS were individually supplemented
– The moisture content 60%; Temp.- 30 ± 2°C; the content was tilled for aeration 3X/week

18. • Determination of total petroleum hydrocarbon
– Hydrocarbon content of the soil samples was determined by Spectrophotometry.
– The total petroleum hydrocarbon (TPH) in soil was estimated using the standard curve
derived from fresh used engine oil diluted with toluene.
• Enumeration and Identification of soil bacteria
– Soil samples from each oil polluted soil were taken every 14 days for the enumeration of
total Aerobic Heterotrophic Bacteria (AHB).
– Hydrocarbon utilizing bacteria (HUB) in the soil samples were enumerated using oil agar.
• incubated at 30°C for 5 days before the colonies were counted.
– The bacterial colonies were randomly picked, and pure culture was obtained.
– The bacterial isolates were characterized using microscopic techniques (Gram staining) and
biochemical tests.

19. Results

20. Fig. 1. Percentage biodegradation of petroleum hydrocarbon in soil
contaminated with used lubricating oil and amended with organic
wastes.

23. Conclusion
• Biodegradation of used lubricating oil was positively enhanced by the
amendment of organic wastes namely CD and SS. Hydrocarbon utilizing bacteria
(HUB) counts were 10% higher in all organic wastes amended soil, compared to
un-amended control soil throughout the period of study.
• Percentage of biodegradation of used lubricating oil in the soil recorded 28%
(CD) and 16% (SS) higher biodegradation compared to control soil without
organic waste amendments.
• Bioremediation can be a viable and effective response to soil contamination with
petroleum hydrocarbons.

24. Waste Water and Industrial Effluents (Case Study)

25. • CETP in Maharashtra at Solapur (capacity- 3 MLD municipal sewage)
– result in to waste water.
• To become water self-sufficient and to meet increasing process water
requirements, the CETP plant realizes the importance of reuse of waste water for
agricultural and industrial uses.

26. Effluent Characteristics from Textile Industry
Process Effluent Composition Nature
Sizing
Starch, waxes, carboxymethyl cellulose (CMC),
polyvinyl alcohol (PVA), wetting agents.
High in BOD, COD
De-sizing Starch, CMC, PVA, fats, waxes, pectins High in BOD, COD, SS, dissolved solids (DS)
Bleaching
Sodium hypochlorite, Cl2, NaOH, H2O2, acids,
surfactants, NaSiO3, sodium phosphate, short cotton fibre
High alkalinity, high SS
Mercerizi
ng
Sodium hydroxide, cotton wax High pH, low BOD, high DS
Dyeing
Dyestuffs urea, reducing agents, oxidizing agents,
acetic acid, detergents, wetting agents.
Strongly coloured, high BOD, DS, low SS, heavy metals
Printing Pastes, urea, starches, gums, oils, binders, acids, Thickeners, cross-linkers, reducing agents, alkali
Highly coloured, high BOD, oily appearance, SS slightly
alkaline, low BOD

27. Flow Diagram of CETP Plant Solapur Maharashtra

28. Technical specification of CETP Unit, Solapur
Units Size Quantity
Screen Chamber 2.0 m x 0.5 m x 0.4 m (LD) 1
Grit Chamber 4.8 m x1.2 m x 1.0 m (LD) 1
Collection/ Equalization Tank 25 m Dia. x 3.0 m (LD) 1
Flash Mixer 2.0 m x1.5 m x 1.2 m (LD) 1
Primary Clarifier 13 m Dia. x 3.0 m (SWD) + 0.3 m (FB) 1
Sludge Holding Tank 4.0 m x 4.0 m x3.0 m (LD) 1
Aeration Tank 25 m x 12 m x4.5 m (LD)+0.5 m (FB) 2
Secondary Clarifier 16 m Dia. x 3.0 m (SWD) + 0.3 m (FB) 1
Chemical Oxidation Tank 7.0 m x 7.0 m x 3.0 m (LD) 1
Pressure Sand Filter 3.2 m Dia. x 1.5 m 1
Activated Carbon Filter 3.2 m Dia. x 1.5 m 1
Treated Effluent Storage Tank 12 m x 12 m x3.0 m 1
LD-Liquid Depth MOC-Made of Concrete Free Board
SWD-Side Water Depth RCC-Reinforced Cement Concrete MS-Mild Steel

29. Effluent Treatment System
Removes coarse solids and other large materials.
Screens/ grates for removal of large materials
Comminutors for grinding of coarse solids
Pre-aeration for odour control
Physical separation of suspended solids from the wastewater
using primary clarifiers.
Reduction of TSS (25-50%) and BOD (50-70%)
Sedimentation chambers, fine screening, flocculation and
floatation used.
Decomposition of suspended and
dissolved organic matter using microbes.
Activated sludge process/ Biological
filtration methods.
Improves wastewater quality before it is reused, recycled or discharged
to the environment.
Used for effluent polishing (BOD, TSS), nutrient removal (N, P), toxin
removal (pesticides, VOCs, metals) etc.

30. Sludge Dewatering System
• Sludge from primary and secondary clarifiers is collected in primary sludge sump.
• From primary sludge sump, sludge is transferred to sludge thickener.
• Thickened sludge is sent to sludge drying beds for removal of water from sludge.
• Overflow from thickener is taken into primary clarifier.
• Thickener collected from sludge dewatering system is collected in Decanter
collection tank.
• That Decanter is then taken into waste water collection tank for further treatment.
• Dried sludge from sludge drying beds is removed, packed and disposed to the
Transport, Storage and Disposal Facility site for secured land filling.

31. Conclusion
• The study indicates that there is efficient reduction in parameter from treatment
units of CETP.
• Up to 20% COD reduction is obtained at biological treatment.
• Removal of oil and grease is also in desirable range.
• pH variations are there in outlet but outlet pH values are in required range.
• Chlorides reduction is not obtained anywhere in the treatment provided at CETP.
• There is need to provide treatment for chloride removal.
Parameters Inlet Outlet
pH 7.9 7.3
COD 900 mg/l 180 mg/l
BOD 600 mg/l 25 mg/l
Oil and Grease 20±5 mg/l 6±3 mg/l
TSS 245 mg/l 80 mg/l
TDS 3300 mg/l 2500 mg/l