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![Biosafety assessment of leachate after biological
treatments
Cytotoxicity
Genotoxicity
Estrogenicity
MTT Assay
Comet Assay
E-Screen
Assay
(Nwagbara et al.
2007)
(Singh et al.
1988)
(Vanparys et al.
2006)
*Huh 7 cell line is used for evaluating cytotoxicity and genotoxicity as hepatocytes express many
nuclear receptor proteins that regulate the expression of xenobiotic metabolizing enzymes like CYP
1A1.
*An estrogen receptive cell line MCF 7 is used for E-Screen assay.
References:1) Nwagbara O, Darling-Reed SF, Tucker A, Harris C, Abazinge M, Thomas RD and Gragg RD. 2007. Induction of cell death, DNA strand breaks, and cell cycle
arrest in DU145 human prostate carcinoma cell line by benzo[a]pyrene and benzo[a]pyrene-7,8-diol-9,10-epoxide. International Journal of Environmental Research and Public
Health.4: 10–14.
2) Singh NP, McCoy MT, Tice RR and Schneider EL. 1988.A simple technique for quantitation of low levels of DNA damage in individual cells.Experimental Cell
Research.175: 184-191.
3) Vanparys C, Maras M, Lenjou M, Robbens J,Van Bockstaele D and Blust R. 2006.Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7
breast cancer cells. Toxicology in Vitro.20:1238–1248.](https://image.slidesharecdn.com/biotechnologicalapplicationsforenvironmentalwastemanagement-140227060009-phpapp01/75/Biotechnological-applications-for-environmental-waste-management-65-2048.jpg)
Uploaded byUtkarsh Verma
Biotechnological applications for environmental waste management
This document discusses biotechnological applications for environmental waste management. It begins by outlining some key environmental issues like global warming, energy and water contamination problems. It then discusses various waste treatment options like bioremediation, phytoremediation and different microbial bioremediation approaches. Finally, it maps out the field of environmental biotechnology, covering areas like toxicology, bioproducts, biosensors, and systems approaches.
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Biotechnological applications for environmental waste management
- 1. Biotechnological applications forenvironmental waste management What is environment? What are remedial methods? Natural Why environmental problems? Technology development? Xenobiotic compounds Emerging toxicants containing waste Feb. 17, 2014
- 2. Major environmental issue Impactof green house gases Formation of dioxin-like compounds in the environments
- 3. Important environmental problems •1. Global warming- GHG (CO2, CH4, N2O) • 2. Energy problem- Bioethanol, biodiesel, Hydrogen how?-a substitute• 3. Water contaminants/toxicants/eutrophication • 4. Soil degradation/solid waste generation • 5. Air pollutants
- 4. Bioremediation- A potentialapproach for clean and Phytoremediation green environment 4.1 T Caviation methods Ec 5. Toxicology- Toxicogenomics & detoxification Reporter Gene nt me n viro r, En Ai l s oi d r an te Wa l tura ic Na iot b e no o un ds X p com try s mi n & e Chtractioysis ts 1. Ex anal utan p of oll 4.2 Microbial Bioremediation in situ & ex situ Prob lar es bios & ensor 2. nism rga roo robial Mic Mic gy & ol o 4. E ngin Mole eering cu Climate/ Climate meteorology 5.Bioproducts & Bio Chemicals biomaterials Process Engineering Biotechnology 3. Molecular BiologyCatabolic Enzymes & genes Bio Environment & Engineering (Environmental Biotechnology) 6. System 6. System approach approach
- 5. Origin of Earthand Environment The Universe created by colossal explosion that we now refer to as the Big Bang and Planets of the solar system The Earth and Environment
- 6.
- 7. Climatic changes onthe Earth Movement of air during rotation of the Earth and formatio n of cells
- 8. PHYSICAL FACTORS ANDBIOTIC & ABIOTIC MATERIALS IN ENVIRONMENT
- 9. Origin of lifeon the Earth
- 10. Classification of livingorganisms Biodiversity
- 11. Classification of microorganisms Classificationof plants and animals
- 12. Biomolecules in organisms • Itis organic compound composed of carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus and sometimes some other elements. • • • • • • Different types of biomolecules are: A. Small molecules mainly include molecules like:Lipids such as phospholipids, glycolipids, sterols, and glycerolipids: Carbohydrates- provide energy and act as energy storage molecules. Vitamins-survival and health of organisms. Hormones, neurotransmitters and metabolites: - metabolic processes and functions. • • B. Monomers include:Amino acids: - building blocks of proteins function as genetic code and as biomolecules, that assist in other processes such as lipid transport. Nucleotides: - Chemical energy (ATP,GTP), assist in cellular signaling, and enzymatic reactions (coenzyme A, flavin adenine dinucleotide, flavin mononucleotide, nicotinamide adenine dinucleotide phosphate etc ). Monosaccharide: - provides energy and are the building blocks of polysaccharides. • •
- 13. Emergence of manand social environment
- 14. • Understanding HumanBehavior and the Social Environment • Natural Resources: Air, water, soil, minerals etc. • Industrial Revolution-1760-1850 onwards
- 15. Environmental degradation • Theten threats identified in 2004 by the High Level Threat Panel of the United Nations are these: • • • • • • • • Poverty Infectious disease Environmental degradation Inter-state war Civil war Genocide Other Atrocities (e.g., trade in women and children for sexual slavery, or kidnapping for body parts) Weapons of mass destruction (nuclear proliferation, chemical weapon proliferation, biological weapon proliferation) • Terrorism • Transnational organized crime
- 16. Major environmental issuesbefore us
- 17. Contd. Pulp and papermill effluent Molasses from sugar cane mill for distillation Petroleum waste 17 million gallon oil spill under the Greenpoint section of Brooklyn Waste dumping grounds in Delhi
- 18. Emerging industrial pollutants Industrialsources Pops in pulp & paper effluent Pops in distillery effluent • Pulp and paper industry lignosulphonic acid, chlorinated resin acid, chlorinated phenols dioxins, dibenzofuran, bipheny chlorinated hydrocarbon Distillery industry melanoidins Pops in tannery effluent Tannery industry Chlorinated phenolics, PCPs, chromium Municipal Plastic, dioxins, antibiotic etc Pops in municipal sludge Transport Metals, organics Incineration and plastics etc.
- 19. Fate of OrganicCompounds in the Environment ENVIRONMENTAL POLLUTANTS AIR Water Soil
- 20. Major conferences andmeetings • United Nations Conference on the Human EnvironmentSweden in 1972: Declaration containing 26 principles concerning the environment and development 6. Pollution must not exceed the environment’s capacity to clean itself 19. Environmental education is essential 20. Environmental research must be promoted, particularly in developing countries The United Nations Conference on Environment and development (UNCED)- Rio Summit and- Earth Summit United Nations Framework Convention on Climate Change-Kyoto Protocol-reduce emissions of greenhouse gases In Doha, Qatar, on 8 December 2012, the "Doha Amendment to the Kyoto Protocol
- 22. Global warming gasesin the environment
- 23. The potential mechanismsthat regulate the responses of GHGs (CO2, CH4 and N2O) Production and consumption to elevated N (ANPP, aboveground net primary productivity; BNPP, belowground net primary productivity; SOC, soil organic carbon; DOC, dissolved 23 organic carbon; DIN, dissolved inorganic nitrogen; DON, dissolved organic nitrogen).
- 24. Climate change andBiodiversity Role of organisms- autotrophic & chemoautotrophic in CO2 mitigation Carbonic anhydrase Biosurfactants Bioscrubbers for CO2 sequestration
- 25. Solid waste generationfrom different sources 1. Garbage- putrescible, heating value 2. Rubbish- Non putrescible, heating value 3. Pathological 4. Industrial Municipal waste 5. Agriculture waste 6. Medical waste 7. Electronic waste Biodegradable Natural waste Non biodegradable waste Xenobiotic Hazardous - ignitable (i.e. flammable), oxidizing, corrosivity, toxic Radioactive, eco-toxic, explosive •Non-hazardous waste
- 27.
- 28. Waste water treatmentoptions Primary treatment Screening Grit removal Equalization Storage Grinders Flocculation Sedimetation Floatation Coagulation Secondary treatment Aerobic Tertiary treatment Anaerobic Activated sludge process Tricking filter Fixed film reactor Rotating reactor Stabilization pond Chemical oxidation Filtration Carbon adsorption Osmosis Electrolysis Cavitations Photodegradation Upflow anaerobic sludge blanket reactor An. Fludized bed reactor Anaerobic lagoons An. Contact reactor An baffled reactors
- 29. Origin of differenttypes of chemical compounds in the environment and their fate
- 30. Significance of lignocellulosics •Total forest cover 3870 million hectares or 30% of the earth’s land area. • 50% 0f all biomass with an estimated annual production of 50 billion tons. • Half of the residues remain unused while some are used as material and energy-green manure and feed for low producing ruminants. • Major substrate for food, feed, energy, and other commercial items. • Degrading enzymes have potency for fuel, chemicals, food, brewery and wine, animal feed, textile and laundry, pulp and paper, agriculture and pharmaceuticals. • Unused biomass is major source of “waste”- pose an environmental pollution problem.
- 31. Lignocellulosic ecosystem :cellulolytic, hemicellulolytic and liglinolytic strains
- 32. Structure of lignocellulose •Cellulose : Made up of linear chains of β-1,4-linked D-glucose residues. • Hemicellulose : Made up of branched heteroglycans with a backbone of β-1,4-linked Dxylopyranosyl residues with branches of α-1,3linked L-arabinofuranosyl and α-1,2-linked 4-Omethyl-glucoronic acid residues. • Lignins is heterogeneous, three dimensional polymer composed of oxyphenyl propanoid units connected by c-c and c-o-c linkages. It is formed by random coupling of coniferyl alcohol, sinapyl alcohol and p-coumaryl alcohol.
- 33. Lignocellulosic components andits importance as biomaterials Lignocellulose Cellulose Pulp Glucose Cellulose Hemicellulose Furfurals Xylose pulp derivatives Fuel Feed and commercial items Single cell proteins Xylitol Lignin Vanillin Gallic acid Phamaceuticals Herbicides Antifoming agents House hold products
- 35. Degradation of celluloseby enzyme cellulase Pre hydrolysis: Acid, Alkali, ammonia Enzymes: Thermolhilic, alkalophilic, multiplicity Products-fuel. feed, food, commercial products Biofuels Applications: Pulp, industries, food, feed, fuel etc.
- 36. Generalized mechanism ofenzymatic cellulose hydrolysis Problems: 1. End product inhibitions Biotechnology 1. Mutants 2.Protoplast fusion 3. Genetic engineering 4. More enzyme 5. Protein engineering 6. Cellulosomemulticomponents enzyme system
- 37. Hemicellulose and degradation-Enzyme xylanase •HC is homo and heteropolymer •AnhydroB-(1,4)D-xylopyrannose, mannopyranose, glucopyranose, galactopyranose •Monomer is D-Xylose Applications 1. Energy 2.Food & feed industries 3. Pulp and paper- Biopulping & biobleaching 4. Waste management 5. Saccharifications of agrowaste 6. Nutritional quality 7. Enhancing texture
- 38. Lignin structure anddegradation Fig. 3: The 1. 2. 3. 4. 5. 6. three common monolignols Prior 1970- no information for degradation 14 C-labelled synthetic lignin Electron microscopy Lignin degrading fungi-White rot, soft rot, Brown rot, other Enzymes Physiological parameters-oxygen, nitrogen, carbon, temp. pH, nutrients
- 39. Involvement of enzymesin degradation of lignin 1. Lignin peroxidase (LiP) Extracellular, H2O2 dependent, glycosylated hemprotein, MW 41-42 kDa, 2. Manganese peroxidase (MnP) Extracellular, H2O2 dependent, MnII-dependent, neutral carbohydrate, MW 41-45 kDa 3. Laccase Extracellular, non-heme, copper containing 4. Other phenol-oxydizing enzymes 5. Glyoxal oxidase Support oxidative turn over of LiP and MnP reduction of O2 to H2O2 with oxidation of substrate Applications 1. Industrial, 2.Commercial, 3. house holds, 4. waste management
- 40. Biodegradation and bioconversionof lignocellulosic waste in the environment Fermentation Bioethanol 1.Cellulases 2.Xylanases 3.Laccase4.Lignin peroxidase & 5.Manganese peroxidase Schematic diagram- ethanol production from sugarcane bagasse
- 44. Biotechnological innovations: biomaterials-biorefinery • Screening for organisms with novel enzymes: enzyme evolution-random mutagenesisrecombination-selection-screening • Strain improvement of existing industrial organisms and enzyme engineering • Production and operation related factors-Process optimization – Substrate – Culture conditions – Recycling of enzymes – Redesigning of processes – Process optimization models and soft wares
- 45. Strain improvement ofexisting industrial organisms and enzyme engineering • Hyper producer organisms • Robust organisms – Culture conditions: isolation of 1% strains-Great culture plate enigma – Biomining through: • Genomics-complete blue print of the organism • Metagenomics-genomics with functional aspects at community level – Necessity of discovering unique gene, cloning, quantitative analysis, and expression
- 46. Process optimization Bioreactors laboratory scale Pilotscale Industrial scale Liquid state Fermentation -Homogeneous -Heterogeneous Stirred tank reactor Air-lift or bubble-column reactor Batch Continuous Fed-batch Solid state Fermentation Flask Tray Packed bed Tunnel Paddle Rotating drum Tower
- 47. • • • Biofuel Production andintegrated pollution control using microalgae Microalgal Farming and CO Mitigation 2 Microalgal Farming using Wastewater Microalgal Farming using Marine Microalgae Possible routes to energy products Basic overview of the pathway of carbon capture and lipid biosynthesis
- 48. Anthropogenic chemical compoundsin environemnt
- 49. Persistent organic pollutantsin environment • Wide distribution- POPs detected from soil, water, food items, commercial products • Sources- Mostly chlorinated organic compounds formed unintentionally- industries, commercial, agriculture, military, other human activities, and natural sources • Insufficient data- No reliable data for their persistence in Indian environment- No management practices • Problems in detection methods- Methods for detection and degradation not up to the mark. • Highly toxic and recalcitrant- ultimate formation oftetrachlorodibenzo-p-dioxin and furan-like compounds-complete physiological impairment. • Tremendous scope for medical diagnostics and therapy and products. • Therefore, methods & technology for detection,
- 50. Degradation of aromaticcompounds
- 51. Key component: POPs Emergingenvironmental contaminant in present scenario
- 52. Classification of POPs •Dirty Dozen - UNEP Stockholm Convention on Persistent Organic Pollutants - 2001 aldrin dieldrin toxaphene chlordane endrin mirex polychlorinated biphenyls heptachlor DDT polychlorinated dibenzo-p-dioxins polychlorinated dibenzofurans hexachlorobenzene • UNEP has added nine new chemicals (all are poly haloginated compounds) to the "dirty dozen" list of restricted or banned toxic chemicals in 2009. • Some other organic pollutants that may be persistent or lead to formation of dioxin like compounds in the environment include: Poly Aromatic Hydrocarbons Aromatic amines Pyrethroids Volatile Organic Compounds Metabolites of VOCs Phthalates
- 53. Biomagnification >Biomagnification, also knownas bioamplification, or biological magnification is the increase in concentration of a substance, such as the pesticide DDT, that occurs in a food chain as a consequence of: Food chain energetics >Low (or nonexistent) rate of excretion/degradation of the substance.
- 54. Persistence and detectionof dioxin-like POPs • Dioxin detected from food items, human exposure, milk and its products, environmental sources from US, Japan and EU countries. • No reliable data from developing countries including India. • Detection methods. • Instrument development. • Thermokinetic modelling, equilibrium modelling, statistical determinations and others. • Field validation. • Laboratory
- 55. Biodegradation strategies forremoval of organic compounds in environment • Possible use of biodegradation processes -Indigenous microorganisms -Genetically modified microorganism -Continuous enrichment of microorganism #Culture dependent and culture independent microorganisms-Metagenomic approach
- 56. Fate of organiccompounds in the uptake into the cells and degradation, assimilation and mineralization
- 57. Degradation of methane Catabolicgene in degradation of alkanes
- 58. Remediation of POPsin waste sites
- 59. What is Bioremediation? •Bio = living • Remediate = to bring the sites and affairs • into the original states • Bioremediation can be defined as any process that uses microorganisms, green plants or their enzymes to return the environment altered by contaminants to its original condition. • Bioremediation technology using microorganisms was reportedly invented by George M. Robinson. • Use of biological sciences and technology for metals and organic compounds remediation.
- 60. Bioremediation Potential alternative forconservation and management of environment Bio = living Remediate = to bring the sites and affairs into the original states Bioremediation can be defined as any process that uses microorganisms, fungi, green plants or their enzymes to return the environment altered by contaminants to its original condition. BIOAUGUMENTATION BIOSTIMULATION Enzymatic methods Ex situ Bioremediation In situ Bioremediation
- 61. Soil–plant–microbial interactions inremediation of pollutants in environment Importance of soil–plant–microbial interactions in bioremediation
- 62. Biocolloid formation inmetal bioremediation Colloidal aggregation–flocculation or attachment to inorganic and organic particles in water can lead to settling and removal of metals from the water column to the bottom sediment
- 63. Technologies in Bioremediation Exsitu bioremediation • • • • • • • • • Electro kinetically enhanced remediation Soil Washing Soil mound Bioxidation ProcessDispersing by Chemical Reaction Biocolloid formation Bioreactors Land Treatment Composting Lagoons (aerobic/ anaerobic) Partial peroxidation In situ bioremediation • • • Bioventing Bioslurping Biopiling
- 64. Limitation of in-situremoved by Enhancement of Bioremediation Use of microorganisms to degrade contaminants in saturated soils and groundwater obtaining harmless chemicals as end products
- 65. Biosafety assessment ofleachate after biological treatments Cytotoxicity Genotoxicity Estrogenicity MTT Assay Comet Assay E-Screen Assay (Nwagbara et al. 2007) (Singh et al. 1988) (Vanparys et al. 2006) *Huh 7 cell line is used for evaluating cytotoxicity and genotoxicity as hepatocytes express many nuclear receptor proteins that regulate the expression of xenobiotic metabolizing enzymes like CYP 1A1. *An estrogen receptive cell line MCF 7 is used for E-Screen assay. References:1) Nwagbara O, Darling-Reed SF, Tucker A, Harris C, Abazinge M, Thomas RD and Gragg RD. 2007. Induction of cell death, DNA strand breaks, and cell cycle arrest in DU145 human prostate carcinoma cell line by benzo[a]pyrene and benzo[a]pyrene-7,8-diol-9,10-epoxide. International Journal of Environmental Research and Public Health.4: 10–14. 2) Singh NP, McCoy MT, Tice RR and Schneider EL. 1988.A simple technique for quantitation of low levels of DNA damage in individual cells.Experimental Cell Research.175: 184-191. 3) Vanparys C, Maras M, Lenjou M, Robbens J,Van Bockstaele D and Blust R. 2006.Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7 breast cancer cells. Toxicology in Vitro.20:1238–1248.
- 66. Molecular Probes fortracking
- 67. Miniaturized ecogenomic sensorsto measure microbial activity-carbon sequestration • The sensors could be installed into advanced ocean observatories to monitor DNA and RNA from diverse microbial communities. • Subsystems for monitoring, data management and communication, and data modelling would be incorporated for data contextualization. • The sensors would report to a worldwide network of laboratories in real time by satellite telemetry. • Culturable and nonculturable (metagenomics) bacteria for degradation of organic compounds & carbon concentrating mechanisms and value added products.
- 68. System biology approaches Thefour-step paradigm for metabolic systems biology
- 69. Conclusion • POP/ DFand its congeners are difficult to detect in the environment. • Degradation of POP/DF in several steps by formation of intermediary metabolites. • Degrading genes are present in various locations. • Bioremediation difficult. • Bioassay methods are useful which may be optimized and developed. • System approach is recent days methods.
- 70.