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Problem 9: Micro-organisms require cellular building blocks, such as (carbon) C, (hydrogen) H, (oxygen) O, (nitrogen) N, (phosphorus) P, and minerals for

growth. These can be obtained through consuming organic substances containing these elements, or from inorganic materials, such as carbon dioxide, water, nitrate and phosphate. Micro-organisms also require energy. They obtain this through respiration. In this process organic carbon is oxidised to release its energy. Oxygen or other hydrogen acceptors is needed for the respiration process. Algae and photosynthetic bacteria can also utilise energy from sunlight, while certain types of bacteria can utilise energy from chemical reactions not involving respiration. The building blocks and energy are used to synthesise more cells for growth and also for reproduction. In the treatment of wastewater three types of overall processes are distinguished to represent the conversion of organic wastes by micro-organisms. The classification is based on whether the environment where the process takes place is aerobic, anaerobic or photosynthetic. Under aerobic conditions (in the presence of oxygen), micro-organisms utilise oxygen to oxidise organic substances to obtain energy for maintenance, mobility and the synthesis of cellular material. Under anaerobic conditions (in the absence of oxygen) the micro-organisms utilise nitrates, sulphates and other hydrogen acceptors to obtain energy for the synthesis of cellular material from organic substances. Photosynthetic organisms use carbon dioxide as a carbon source, inorganic nutrients as sources of phosphate and nitrogen and utilise light energy to drive the conversion process. Micro-organisms also produce waste products, some of which are desirable and some undesirable. Gases such as carbon dioxide and nitrogen are desirable, since they can be easily separated and do not produce pollution. Gases such as hydrogen sulphide and mercaptans, although easily separated require treatment for odour. Micro-organisms' cellular materials are organic in nature and can also cause pollution. It would be desirable if the cellular materials have undergone self oxidation (endogeneous

respiration utilising own body cells) to produce non-biodegradable materials that are relatively stable. Self-oxidation is achieved when there is no substrate/food available.

A. Municipal Wastewater Treatment

Municipal Wastewater treatment processes use microbes for treatment. Microbes involved in nitrifying processes, anaerobic ammonia oxidation processes and methane fermenting processes are studies carefully for this purpose. The secondary treatment for wastewater involves the use of biological methods that approximate natural degradation processes. It sometimes includes chlorination to accomplish chemical oxidation and disinfection. In biological purification, the microbes bring about the following changes: Coagulation and flocculation of colloids and pseudocolloids Oxidation of carbonaceous matter to CO2

Nitrification i.e. oxidation of NH3 (derived from breakdown of nitrogenous organic matter to nitrite and then to nitrate) The two main processes involved are trickling/percolation filters: these are manufactured beds of crushed stone used to percolate the waste water. They consist of circular or rectangular beds, about 1.8m deep of well-graded media. (clinker,stone,gravel,coke or even coal) usually about 40mm in size but increasing to 100-150mm for bottom enclosed in walls of brick or concrete. The filters bring the wastewater into contact with air and with the biological slime on the filter. The slime contains bacteria mainly zoogloea, fungi, protozoa, especially ciliates and other living organisms. The effective zone for aerobic bacterial activity is 2-3mm below the surface of the slime and their decomposition by the bacteria and fungi in the slime. Absorption and adsorption of solid and suspended matter takes place from the wastewater into and onto the slime. The slimes develop and proliferate on the surfaces of filtering medium over which the water trickles. This is followed by decomposition and synthesis of both the soluble substances and sludge forming solids. Near the top of the trickling filter, aerobic growth is important but further down, where less atmospheric oxygen diffuses, anaerobic action becomes predominant. When the slime layers become very thick, they slough off and can be removed. A large number of scouring animals like worms, flies and larvae are present in the filters which fed upon the biological film of slime preventing its overgrowth. The capacity of a filter can be increased by recirculation. This usually involves the biological filtration of settled sewage, which has been diluted, with a proportion of settled purified effluent from the filters. OXIDATION PONDS: Also known as stabilization ponds, or lagoons, these are used for the simple secondary treatment of sewage effluents in rural communities and some industries. Such ponds are inexpensive to build and operate but require large areas of land. Generally, a 2 stage operation ensues. The 1st stage is analogous to primary treatment; the sewage pond is deep enough so that conditions are anaerobic. Sludge settles

down in this stage. In the 2nd stage, which roughly corresponds to secondary treatment; effluent is pumped into an adjoining pond/s shallow enough to be aerated by wave action. Heterotrophic bacteria (organisms requiring organic compounds for growth and reproduction, the organic compounds serve as sources of carbon and energy) degrade sewage organic matter within the ponds, producing cellular material and mineral products that support the growth of algae. The proliferation of algal population in these lagoons produces oxygen that replenishes the oxygen depleted by the heterotrophic bacteria, permitting continued organic matter decomposition. Because oxygenation is usually achieved by diffusion and by the photosynthetic activity of algae, such ponds need to be shallow. Typically, these are less than 10 feet deep which maximizes the euphoric zone for algal growth. The degradation of organic matter in these ponds is relatively slow and residence time for the treatment of domestic sewage maybe as long as a week. If properly operated the ponds are reasonably free from bad smells, possibly due to deodorizing effect of the chlorophyll in the algae. Treatment of sewage in such ponds usually gives a good reduction in BOD. Algae from these ponds are rich in proteins and fat may be used as cheap animal feed.

B. Hazardous Waste Treatment

Metals are common contaminants worldwide. Long-term deposition of metals in soils can lead to accumulation, transport and biotoxicity/zootoxicity caused by mobility and bioavailability of significant fraction of the metals. Contaminant bioavailability is increasingly being used as a key indicator of potential risk that contaminants pose to both environmental and human health. However, the definition of bioavailability and the concepts on which it is based are still unclear, the methods adopted for its measurement vary and as such there is no single standard technique for the assessment of either plant availability of contaminants or their ecotoxicological impacts on soil biota. Moreover,

bioavailability is often assumed to be static in nature where most decisions on risk and remediation are based on laboratory estimations of the bioavailable fraction, which may vary with time, nature of species as well as with temporal variation in environmental factors. Because of their immutable nature, strict natural attenuation processes alone may not be sufficient in mitigating the risks from metals. However, accelerating these processes with human interference (i.e., assisted natural remediation) that effectively immobilizes metals might be a viable option. Application to soils of certain amendments that enhance key biogeochemical processes in soils that effectively immobilize metals have already been demonstrated in Europe and North America on a field scale. Case studies using lime, phosphate and biosolid amendments have demonstrated, under field conditions, enhanced natural remediation resulting in substantially improved vegetation growth, invigorated microbial population and diversity, and reduced offsite metal transport. Depending on soil/hydrogeochemical properties, source term and metal form/species, and land use, the immobilization efficacy induced by such assisted natural remediation may be enduring. The use of green plants as a remediation tool in environmental cleanup has also offered some potential. Plants can uptake and bioaccumulate (phytoextraction) as well as immobilize (phytoimmobilization) certain trace elements, in conjunction with their rhizospheric processes. While long-term stability of certain metal complexes, such as metal pyromorphites has been shown in model systems, the influence of plant roots and its microbial and mycorrhizal association on such stability is unknown. A suite of chemical and biological tests are available to monitor the efficacy of assisted natural remediation.

C: Soil Remediation Natural soil remediation is biodegradation where trillions of microbes already in the soil eat billions of tons of dead and decaying organic matter every year and convert it into non-organic soluble or gaseous compounds. These compounds in turn take part in further soil processes to form humus that provides nutrients to plant life. Collectively these processes to form humus are referred to as mineralization.

Importantly, some of these soil microbes (approximately 20%) also consume the hydrocarbons commonly found in oil and petroleum products. When a spill occurs, these hydrocarbon-eating microbes begin to emit enzymes to break down the contaminants, releasing only water and carbon dioxide as waste. They literally have a feast and are able to quickly increase in number.

However, because of the sudden increase of populations, critical nutrients for growth in the soil like carbon (in the form of smaller carbon chains), nitrogen and phosphorus quickly become depleted. These elements along with hydrogen and oxygen are the main elements required for reproduction, and without these nutrients, the growth of the microbial population is impeded. The microbes become unhealthy and begin to die off, leaving the contaminating hydrocarbons in the soil virtually untouched. D. Groundwater Remediation

Bioremediation generally requires a mechanism for stimulating and maintaining the activity of these microorganisms. This mechanism is usually a delivery system for providing one or more of the following: An electron acceptor (oxygen, nitrate); nutrients (nitrogen, phosphorus); and an energy source (carbon). Generally, electron acceptors and nutrients are the two most critical components of any delivery system.

In a typical in-situ bioremediation system, groundwater is extracted using one or more wells and, if necessary, treated to remove residual dissolved constituents. The treated groundwater is then mixed with an electron acceptor and nutrients, and other constituents if required, and re-injected upgradient of or within the contaminant source. Infiltration galleries or injection wells may be used to re-inject treated water. In an ideal configuration, a "closed-loop" system would be established. All water extracted would be reinjected without treatment and all remediation would occur in situ. This ideal system would continually recirculate the water until cleanup levels had been achieved. If your state does not allow re-injection of extracted groundwater, it may be feasible to mix the electron acceptor and nutrients with fresh water instead. Extracted water that is not re-injected must be discharged, typically to surface water or to publicly owned treatment works (POTW).