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- 1. Rishav Prakash (1MV15BT026) VIIIth Semester Guides RManjunath Ishwar Chandra Sir M Visvesvaraya Institute of Technology, Bangalore
- 2. Trace amounts ofheavy metals are required by living organism including copper, iron, zinc but however excessive levels of these metals can be toxic to the organism due to their toxicity and accumulation behavior(Franke S, Grass G, Rensing C, Nies DH (2003) )
- 3. Sources of HeavyMetals Pacyna JM, Pacyna EG (2001) An assessment of global and regional emissions of trace metals to the atmosphere from anthropogenic sources worldwide. Environmental Reviews 9: 269- 298. Czekalski N, Gascón Díez E, Bürgmann H (2014) Waste-water as a point source of antibiotic-resistance genes in the sediment of a freshwater lake. ISME J 8: 1381-1390. 8. Duruibe J, Ogwuegbu M, Egwurugwu J (2007) Heavy metal pollution and human biotoxic effects. International Journal of Physical Sciences 2: 112-118. Sources of heavy metal Mining Agriculture and Farming Solid Waste Automobile spear parts Emission of fossil fuels gases Batteries and E-waste
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- 5. Basic concepts and terminology Biosorptionpassive sequestration by non-metabolizing non-living biomass Bioaccumulation Metabolically mediated transport and deposition of chemical species in living cells Adsorption Involves the interface accumulation or concentration of substances at a surface or interface Sorption + Absorption Molecules or atoms of one phase interpenetrate among other of another phase to form a «solution» Davis TA, Volesky B, Mucci A. Water Research. 2003
- 6. Properties of heavymetals • They occur near the bottom of the periodic table • Have high densities • Toxic in nature • Nondegradable Note: Arsenic is not actually a metal but is a semimetal i.e. its properties are intermediate between those of metals and nonmetals.
- 7. • It isparticularly the cell wall structure of certain algae, fungi and bacteria which was found responsible for this phenomenon (Volesky, 1986) • Bioaccumulation is nothing but a biosorption but it happens in a living cells Biosorption is a property of certain types of inactive, dead, microbial biomass to bind and concentrate heavy metals.
- 8. Mechanism Involved inBiosorption
- 10. Non-living biomass Adsorption amines thioethers carboxylates hydroxyls thiols phosphates amides Livingbiomass Biological Processes Reduction, oxidation, methylation… + Adsorption Transport through the membrane Cellular membrane binding Biosorption processes
- 11. Biosorption of zincion: a deep comprehension ;Vishal Mishra (2013)
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- 13. Transport across cellmembrane transport across microbial cell membranes may be mediated by the same mechanism used to convey metabolically important ions such as potassium, magnesium and sodium. Basically biosorption by living organisms comprises of two steps. 1. Metabolism independent binding where the metals are bound to the cell walls and 2. Metabolism dependent intracellular uptake, whereby metal ions are transported across the cell membrane. ( Costa, et.al., 1990, Gadd et.al., 1988, Ghourdon et.al., 1990, Huang et.al., 1990., Nourbaksh et.al., 1994)
- 15. Physical adsorption • Inthis category, physical adsorption takes place with the help of van der Waals' forces. – Kuyucak and Volesky 1988, hypothesized that uranium, cadmium, zinc, copper and cobalt biosorption by dead biomasses of algae, fungi and yeasts takes place through electrostatic interactions between the metal ions in solutions and cell walls of microbial cells. • Electrostatic interactions have been demonstrated to be responsible for copper biosorption by bacterium Zoogloea ramigera and alga Chiarella vulgaris (Aksu et al. 1992), for chromium biosorption by fungi Ganoderma lucidum and Aspergillus niger .
- 16. Ion Exchange Cell wallsof microorganisms contain polysaccharides and bivalent metal ions exchange with the counter ions of the polysaccharides. o Kuyucak and Volesky 1988: the alginates of marine algae occur as salts of K+, Na+, Ca2+, and Mg2+. These ions can exchange with counter ions such as CO2+, Cu2+, Cd2+ and Zn2+ resulting in the biosorptive uptake of heavy metals o Muraleedharan and Venkobachr, 1990: The biosorption of copper by fungi Ganoderma lucidium and Aspergillus niger was also up taken by ion exchange mechanism.
- 17. Complexation removal cantake place by complex formation on the cell surface after the interaction between the metal and the active groups. Metal +Active group---- complex Aksu et al. 1992 hypothesized that biosorption of copper by C. vulgaris and Z. ramigera takes place through both adsorption and formation of coordination bonds between metals and amino and carboxyl groups of cell wall polysaccharides. Microorganisms + metals -----chelate metallo-organic (citric, oxalic, gluonic, fumaric, lactic and malic acids) Metals may be biosorbed or complexed by carboxyl groups found in microbial polysaccharides and other polymers.
- 18. Precipitation Precipitation maybe either dependent cellular metabolism or independent of it. In the former case, the metal removal from solution is often associated with active defense system of the microorganisms. They react in the presence of a toxic metal producing compounds, which favour the precipitation process. In the case of precipitation not dependent on the cellular metabolism, it may be a consequence of the chemical interaction between the metal and the cell surface.
- 19. it affects thesolution chemistry of the metals, the activity of the functional groups in the biomass and the competition of metallic ions (Friis and Myers-Keith, 1986, Galun Temperature seems not to influence the biosorption performances in the range of 20- 35 0C (Aksu et al. 1992) Biomass concentration in solution seems to influence the specific uptake: for lower values of biomass concentrations there is an increase in the specific uptake (Fourest and Roux, 1992; Gadd et al. 1988). Gadd et al. 1988 suggested that an increase in biomass concentration leads to interference between the binding sites. Fourest and Roux, 1992 invalidated this hypothesis attributing the responsibility of the specific uptake decrease to metal concentration shortage in solution.
- 20. Bacteria as Biosorbents •Bacterial biosorption is mainly used for the removal of metals ions and dyes. Leitão AL (2009) Potential of Penicillium species in the bioremediation field. Int J Environ Res Public Health 6: 1393-1417. . Li X, Li A, Long M, Tian X (2015) Equilibrium and kinetic studies of copper biosorption by dead Ceriporia lacerata biomass isolated from the litter of an invasive plant in China. J Environ Health Sci Eng 13: 37. Cho DH, Kim EY, Hung YT (2010) Heavy metal removal by microbial biosorbents Environmental Bioengineering Springer 11: 375-402. Lone MI, He ZL, Stoffella PJ, Yang XE (2008) Phytoremediation of heavy metal polluted soils and water: progresses and perspectives. J Zhejiang Univ Sci B 9: 210-220. . Green-Ruiz C, Rodriguez-Tirado V, Gomez-Gil B (2008) Cadmium and zinc removal from aqueous solutions by Bacillus jeotgali: pH, salinity and temperature effects. Bioresour Technol 99: 3864-3870.
- 21. Algae as Biosorbents •Algae are efficient and cheap biosorbents – algae absorb about 15.3% - 84.6% which is higher as compared to other microbial biosorbents. • Brown algae – known to have high absorption capacity. Biosorption of metal ions occurs on the cell surface by means of ion exchange method. – absorbed metals like Cd, Ni, Pb through chemical groups on their surface such as carboxyl, Sulfonate, amino, as well as sulfhydryl.
- 23. Fungi as Biosorbentsof Heavy Metals • The use of fungi as bio sorbents material has been shown to be efficient material is also one of the cost-effective and eco-friendly methods with serves as an alternative to chemically bound treatment process. • The capability of the many type of fungi to produce extracellular enzymes for the assimilation of complex carbohydrates for former hydrolysis makes capable the degradation of various degrees of pollutants. • They also have the benefit of being relatively uncomplicated to grow in fermenters, therefore being appropriate for large scale production. • Another benefit is the easy separation of fungal biomass by filtration because of its filamentous structure. In comparison to yeasts, filamentous fungi are less sensitive to variations in nutrients, aeration, pH, temperature and have a lower nucleic content in the biomass
- 25. Biosorption of heavymetals by obligate halophilic fungi Highlights First report of using obligate halophilic fungi for biosorption of heavy metals. Cadmium, copper, ferrous, manganese, lead and zinc were effectively removed from their medium by obligate halophilic fungi. A. flavus and S. halophilus showed best performance for the biosorption of heavy metals. Over all, Fe and Zn were most removed by obligate halophilic fungi. This study provides a cost effective solution of removing heavy metals Javaid Hussain , Chemosphere 199 (2018) 218-222
- 26. Abstract The presence ofheavy metals in the environment poses a serious threat to human health. Remediation of this problem using microorganisms has been widely researched to find a sustainable solution. Obligate halophilic fungi comprising Aspergillus flavus, Aspergillus gracilis, Aspergillus penicillioides (sp. 1), Aspergillus penicillioides (sp. 2), Aspergillus restrictus and Sterigmatomyces halophilus were used for the biosorption of cadmium, copper, ferrous, manganese, lead and zinc. The metals were supplemented as salts in potato dextrose broth for the growth of obligate halophilic fungi and incubated for 14 days. The supernatant and biomass were obtained by the acid digestion method. The biosorption was screened by atomic absorption spectroscopy. All tested fungi showed moderate to high adsorption of heavy metals, amongst which A. flavus and S. halophilus showed the best average adsorption of all heavy metals studied, with an average of 86 and 83%, respectively. On average, Fe and Zn are best removed from the liquid media of obligate halophilic fungi, with an average of 85 and 84%, respectively.
- 27. Materials and methods Aspergillusrestrictus Aspergillus pencillodes Aspergillus gracilis A.flavus (5%Nacl) Potato broth Agar Or Sterigmatomyces halophilus
- 28. Innoculated in aseries of Erlenmeyer flask 50 ml of PDB Initial concentration of heavy metal -1000 ppm Heavy metal used - 𝐶𝑑𝐶𝑙2. 𝐻2 𝑂, 𝐶𝑢𝐶𝑙2. 2𝐻2 𝑂 𝐹𝑒(𝑁𝐻4)2 𝑆𝑂4. 6𝐻2 𝑂, 𝑀𝑛𝐶𝑙2, 𝑃𝑏(𝑁𝑂3)2, 𝑍𝑛(𝑁𝑂3)2 Flask was incubated for 14 days at lab Control medium having no fungal presence Fungal were ontained from media, using Whatman Filter Paper Dried at oven 80© for 12 hrs(jack etal,2005)
- 29. Acid Digestion Protocol 0.15mg of dried sample placed in 100 ml Erlenmeyer flask 20 ml of 65% HN𝑂3 + 3 ml Deionized water 24 hrs, heated on hot plate @95-105© Further heating adding 35% HCl The sample digested by refluxes After cooling filtered through ash less filter paper.
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- 36. Result Heavy Metal Speciesshowing maximum or absorption Cadmium Aspergillus flavus, Sterigmatomyces halophilus Copper A.flavus , A.restrictus ,S.halophilus Iron A.restrictus (least) Manganese A.flavus , S.halophilus Lead A.restrictus, S.halophilus Zinc A.grailis, S.halophilus(90%)
- 37. Inference It wasobserved that out of six species three species shows excellent absorption property for heavy metal such as zinc lead , cadmium Species such as Aspergillus flavus , A.restrictus, Sterigmatomyces halophilus In that also Sterigmatomyces halophilus showed absorbance in every heavy metal In case of Zinc Sterigmatomyces halophilus absorbed 90% of heavy metal.
- 38. Mechanism of Pb2+removal from aqueous solution using a nonliving moss biomass Highlights The moss plant Barbula lambarenensis (RBL) will be useful for metal adsorption from aqueous solution. " The maximum adsorption capacity (Qo) for Pb adsorption is 62.50 mg/g at 298 K and 90.91 mg/g at 323 K. " The pH for optimum adsorption is 5.0 while maximum adsorption was attained in 30 min. " The adsorption data obeyed the pseudo-second-order model. " The free energy changes (DGo) are positive and the reaction is exothermic. Bamidele I. Olu-Owolabi, Paul N. Diagboya, William C. Ebaddan ,Chemical Engineering Journal (2012), 270–275
- 39. Protocol • Sampling, pre-treatmentand characterization of biosorbent material The B. Lambarenensis was obtained in August 2011 in Agbor(611060 0E; 615090 0N), Delta State, Nigeria; washed severally withtap water and deionized water to remove impurities, then air-dried,ground, sieved through a 0.5 mm mesh size sieve, and the sieved particles were used for the study. This was called the raw BL
- 40. • 1000 mg/Lof standardized Pb2+ stock solution was prepared from the analytical grade chloride salt. Working solutions of required concentrations were prepared from this stock as required. • Replicate batch experiments were used to determine metal sorption capacity of the biomass in 60 mL polyethylene bottles by contacting approximately 0.1 g of the RBL biomass with 20 mL of metal solution, except where otherwise stated, for determining effect of pH, time, temperature and sorbate concentration. • The sorbent–sorbate mixtures were shaken on a mechanical shaker duringthe course of the adsorption experiment and concentrations of metal in the filtrate solutions were determined using the Buck Scientific 205 Atomic Absorption Spectrometer (AAS) with air- acetyleneflame on absorbance mode.
- 41. FT-IR test spectraof unloaded RBL biomass showed several absorption peaks between the scanning frequency range of4500–500 cm1
- 42. The water contentin the RBL biomass and the numerous free hydroxyl groups in the polysaccharide structure of this moss plant wall may explain the presence of the band. The sharp absorption just below 3000 𝑐𝑚−1 is indicative of CAH stretch likely from alkanes while the band at around 2350 𝑐𝑚−1 is suggestive of cumulative double bonds stretch of The band around 1700 𝑐𝑚−1 is characteristic of the stretch of carbonyl double bond either from free or esterified carboxyl groups. However, closer band frequencies at 1655 𝑐𝑚−1 have been attributed to amide-I of protein secondary structures. The absorption peak at about 1424 𝑐𝑚−1 suggests aromatic methyl group/methyl ketone and carboxylate vibrations or likely CAH deformations of alkanes. The strong band around 1090 cm1 is due likely to the bond and stretching, which are characteristic for polysaccharides. Peaks below 1000 𝑐𝑚−1 have been attributed to such groups as aromatic bending vibrations (874 𝑐𝑚−1), thioesters (672 𝑐𝑚−1), but majorly plane deformations. The broad band positioned around 3400 𝑐𝑚−1 is indicative of the stretching band of the carbonyl double bond from some carboxylic acids and their salts, a stretching vibration of free hydroxyl functional groups of aromatic and aliphatic origins, and possibly NH stretch of amides
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- 44. The uptakeof Pb2+ ions is pH dependent. At the low pH value of 3.0, RBL adsorbed lower amount (56.4 mg/g or 57.8%) of Pb2+ onto its surface active adsorption sites. With increase in solution pH, the quantity adsorbed increased steadily until pH 5.0 (70.6 mg/g or 71.6%) and 6.0 (72.4 mg/g or 73.3%) where the quantities of Pb2+ adsorbed made a plateau on the graph before precipitation of Pb2+ ions set in. In summary, steady increment in adsorption was observed from pH 3.0 to 5.0, indicating that more Pb2+ ions were adsorbed on RBL surfaces as pH increased. Then adsorption reached a plateau between pH 5.0 and 6.0. The optimum pH for adsorption of Pb2+ was recorded at pH 5.0. Above this pH, there was an apparent increase in adsorption of Pb2+. This has been attributed mainly to solvation and hydrolysis of Pb2+ ion products leading to precipitation of Pb2+ from solution. In aqueous solutions of pH less than 5, Pb2+ ions exist as either Pb2+ or Pb(OH)+ or both. However, the formation of Pb2+ hydrolysis products begins to occur at pH values between 5 and 6, and this brings about precipitation. Due to this reason all the experiments were carried out at pH 5.
- 45. Effect of timeon Pb2+ adsorption
- 46. Effect of varyingbiosorbent dose on Pb2+ adsorption.
- 47. Effect of initialadsorbate concentration on Pb2+ adsorption.
- 49. The abilityof B. lambarenensis biosorbent to remove Pb2+ from aqueous solution was investigated in equilibrium, kinetics and thermodynamics studies. The results obtained show that the biomass of B. lambarenensis has an optimum pH for Pb2+ adsorption at 5.0, nearly attains maximum adsorption within 1 h at 298 K, and obeys the pseudo-second order kinetics with an exothermic reaction. Pb2+ adsorption increased with increase in temperature. The Langmuir isotherm described the equilibrium data better than other isotherms indicating adsorption is monolayer with monolayer adsorption capacity of 62.50 mg/g at 298 K and 90.91 mg/gat 323 K. The FT-IR analysis showed that possible functional groups responsible for metal adsorption are carboxyl, carbonyl, amides, hydroxyl and possibly other smaller groups that cannot be identified using only FT- IR spectra.
- 50. The satisfaction andeuphoria that accompany the successful completion of any seminar should be, but incomplete without the mention of the people who made it possible. We sincere thank to our guide Mr. R Manjunath and Mr.Ishwar Chandra (Assistant Professor, Sir M. Visvesvaraya Institute of Technology, Bangalore) for his constant guidance, support and encouragement in conceptualizing, developing and executing the presentation. I also thank Dr. Mrinalini menon for her constant support and encouragement. We express our gratitude to Dr. H. G. Nagendra (HOD, Sir M. Visvesvaraya Institute of Technology, Bangalore) for giving us the opportunity to present such an enriching topic. Our heartfelt thanks to the entire faculty of Sir M. Visvesvaraya Institute of Technology for enabling us with the necessary knowledge, skills and capabilities to present this topic successfully. ACKNOWLEDGEMENT
- 52. Extra Slides • Potatodextrose broth and agar were prepared using 250 g of potato boiled in 100 ml of distilled water for 30 min and the filtrate is mixed with 2 g of dextrose and for agar plates 1.5 g of agar was added with this mixture. • Extreme or obligate halophiles require high osmotic pressure (up to 30% salt) Facultative halophiles tolerate high osmotic pressure.