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INTRODUCTION A wide variety of different biological treatment processes are used but the same basic concepts apply to all of them. Emphasis in this paper is on fundamentals of biological treatment - not on the unique features of specific treatment techniques. Biological treatment has been applied primarily to organic wastewater although there are other applications. In biological treatment or organic wastewater, the same transformations that would occur naturally in receiving waters (with associated environmental quality degradation) are caused to occur under controlled conditions in biological treatment facilities. To effectively treat wastes biologically, it is necessary to create conditions that favour growth of desired organisms. To do this, it is necessary to have an understanding of the nature of the organisms involved and their requirements for growth.


NATURE OF_MICROORGANISMS Principal Types Size o Bacteria - about 1 micron Bacteria Blue-green algae Algae Fungi Protozoa

Uniquitous - if conditions favouring their growth are created; they will be in plentiful supply. Temperature Dependance

Roughly, the rate of microbial reactions doubles with a 100C temperature increase. The rate at temperature T is often expressed as RT = R20 (T-2) (1)

Where R20 is the rate at 200C, and is an empirical constant with typical values in the range of 1.02 to 1.08 (METCALF and Eddy, 1979)


Growth Bacteria reproduce by fission Can do so in as short a time as 20 minutes (under extremely favourable conditions in an aerobic environment) Anaerobic organisms growth slower than aerobic Tremendous growth potential limited only by the availability of required nutrients

3.0 3.1


If microorganisms are to function effectively in wastewater treatment plants all ingredients required for their growth must be present. provided. If required nutrients are not found in wastewater being treated, they must be A listing of specific nutrient requirements follows. Energy Source Two types:



Algae, blue green algae, and some bacteria Complex reactions using suns energy such as light H20 + C02 02
+ (CH20)


where (CH20) represents the synthesis new cellular material.

The oxygen produced in Equation 2 may be of use (as an electron acceptor see section 3.3) in biological processes such as oxidation ponds. Note, however, the algae continue to use oxygen during periods of darkness (using the reverse of the reaction in Equation 2).

Chemosynthetic Organisms Bacteria, fungi and protozoana Obtain energy by oxidation of chemical compunds


Two types: Heterotrophs Autotrophs oxidize organic compounds to obtain energy oxidize inorganic substances (for example Fe++, NH4, or H2S) to obtain energy.

While biological wastewater treatment ordinarily involves heterotrophic organisms, opportunities exist for use of autotrophic organisms as well.


Carbon Source Carbon is a major constituent of the microbial cells synthesized in biological wastewater treatment processes. Thus, an abundant source of carbon must be available. This is no problem in treatment of wastewater containing organic compounds the organic contaminants serve as the carbon source for heterotrophic organisms. Autotrophic microorganisms use inorganic carbon (bicarbonate ions or carbon dioxide). In some cases, carbon might have to be added. Nitrogen and phosphorus are significant constituents of cells. (For example, nitrogen is needed for amino acid synthesis and phosphorus is essential for the genetic material, deoxyribonucleic acid). Nitrogen and phosphorus requirements depend on the net amount of synthesis which occurs in biological wastewater treatment processes. Thus, requirements depend on design of the treatment process and the electron acceptor. As a crude guideline, the necessary weight ratio of BOD:N:P is about 100:5:1 in aerobic processes. Many industrial wastewaters are deficient in nitrogen and/or phosphorus, and nutrients must be added to accomplish effective biological wastewater treatment.


Nitrogen and Phosphorus Sources Nitrogen and phosphorus are significant constituents of cells. (For example, nitrogen is needed for amino acid synthesis and phosphorus is essential for the genetic material, deoxyribonucleic acid). Nitrogen and phosphorus requirements depend on the net amount of synthesis which occurs in biological wastewater treatment processes. Thus, requirements depend on design of the treatment process and the electron acceptor. As a crude guideline, the necessary weight ratio of BOD:N:P is about 100:5:1 in aerobic processes.



Many industrial wastewaters are deficient in nitrogen and/or phosphorus, and nutrients must be added to accomplish effective biological wastewater treatment.

Trace Minerals In addition to C, H, O, N and P, microbes require a wide variety of inorganic materials for their metabolism and for synthesis of new cells. For example, the catalytic activity enzymes are often associated with Co, K, Mn, Zn, Cu, Fe, Mo, Zn, etc. Required amounts are small but, still, the trace minerals are essential. Orgindarily assumed to be contained in the wastewater carriage water. But could limit treatment effectiveness.


Growth Factors Some organisms can synthesize all of the organic compounds they require, but others (like humans) requires some preformed organic compounds. Examples are some amino acids, vitamins, etc. Presumably, in heterotrophic systems, lysis of organisms results in availability of necessary growth factors for other organisms.


Water Microorganisms use substances in solutions - thus water is essential. No problem of availability in industrial wastewater treatment systems, but the moisture content of soil can limit the rate of microbial transformations in land treatment systems.


RATE OF SUBSTRATE UTILIZATION Substrate (wastewater constituents) removal in biological processes occurs because of enzymatically catalyzed reactions. Thus, questions on the rate of removal of wastewater constituents are answered by considering the rate at which enzymatically catalyzed reactions occur. Conceptually, it is considered that such reactions occur as follows: E+S ES E+P (4)

where E denotes a specific enzyme, S, the substrate, P the product of the reactions, and ES a temporary enzyme-substrate complex. As in the nature of catalysts, the enzyme is released when the product is formed so as to combine again with new substrate. The maximum rate of an enzymatically catalyzed reaction occurs when all enzymes is in use (that is, it is combined as the enzyme-susbtrate complex, ES).


Use of the law of mass action and Equation 4 leads to the following expression for d(S)/dt, the rate of removal of substrate. d (S) dt = Vm (S) Km + (S) (5)

The relationship between substrate concentration and rate of substrate removal as given in Equation 5 is shown in Figure 1.

When the substrate concentration is high, the rate of removal becomes independent of substrate concentration and approaches the maximum possible rate, Vrn (Zero order kinetic). When the substrate concentration is low, the rate of substrate removal becomes directly proportional to substrate concentration. (First order kinetics). Note that this is the assumption used in developing the traditional BOD rate equation.

Experimental data for pure substrates indicate that values of Km and Vm are such that zero order kinetics prevails until low substrate concentrations are reached. Note the zero order curves in Figures 2 (from ECKENFELDER (1980)). Possibly the first order BOD removal kinetics observed with real wastewaters are the results of summation of many zero order curves from the components of the heterogeneous wastewater.


SLUDGE PRODUCTION IN BIOLOGICAL WASTEWATER TREATMENT Microorganisms in biological wastewater treatment systems oxidize substrate in order get energy required to synthesize more organisms. That is, it is their goal to produce excess sludge. A certain amount of energy is required for cell maintenance and this causes a decrease in net synthesis. This reduction in cell mass is called endogenous respiration. Net synthesis may be expressed as dx = Y dt where x dx dt Y = = = = d(S) - KdX dt concentration of microoganisms rate of change of x with time yield coefficient mass of synthesis/ mass of substrate utilized (6)

Kd = Endogenous decay coefficient The magnitude of the yield coefficient, Y, depends on the nature of the substrate and the electron acceptor. A typical value is 0.5mg cells/mg BOD destroyed. For pure substrates, it is possible to estimate Y from


thermodynamics consideration (CHRISTENSEN and McCarty, 1975). The value of Y can be estimated from experimental observations as well. The magnitude of the endogenous decay coefficient typically is in the order of 0.05 to 0.1 day-1. Note from Equation 6 that if a larger population of organisms, X, is used to achieve the same rate of substrate utilization, d(S)/dt, then net synthesis will be reduced.


OXYGEN REQUIREMENTS IN AEROBIC SYSTEMS The amount of oxygen required for aerobic treatment of organic wastewaters cannot be ascertained from the change in oxygen demand between the influent and the effluent because some of the organic matter removed remains in the form of microorganisms. Taking the oxygen equivalent of microbial cells as 1.42 gm 02/gm cells*, then the amount of oxygen required in aerobic biological waste treatment can be calculated as O2 = BODL - 1.42 X where O2 = daily mass of oxygen required total ultimate BOD removed /day mass of excess biological sludge wasted /day (8)

BODL = X =

* If the approximate chemical composition of cells is taken as C5H7N02, then, the amount of oxygen required to oxidize the cells maybe calculated from the equation: C5H7N02 + 5 O2 7.0 5 CO2 + 2 H2O + NH3 (7)

MEAN CELL RESIDENCE TIME AS A MEASURE OF PROCESS PERFORMANCE By writing mass balance equations on biological reactors operating at steady state, the relationships between wastewater characteristics, process design and process performance can be explored. The reader is referred to LAWRENCE and McCarty (1970) for details. The principal design and operational variable that emerges from the analysis is the mean cell residence time, c The mean cell residence time is the average time a cell remains in the treatment system. The mean cell residence time may be written as: c Where V X = = = VX QuXu + QeXe Volume of the system Concentration of cells in the system (9)


Qu Xu Qe Xe -

= = = =

Rate of sludge wastage Concentration of cells in waste sludge Effluent flow rate Concentration of cells in the effluent

The relationship between the mean cell residence time c and the effluent substrate concentration is: S = Km (1 + Kdc) c (Vm Y-Kd) 1 (10)

The form of Equation 10 is illustrated in Figure 3. No substrate removal occurs at c values below that value at which organisms that can use the substrate are washed out of the system faster than they can reproduce.


REFERENCE CHRISTENSEN, D.R., and McCarty, P.L. (1975), Multi Process Biological Treatment Model", Journal Water Pollution Control Federation, 47, 2652. ECKENFELDER, W.W. Jr. (1980), Principles And Practice of Biological Wastewater Treatment, Proceedings of Summer Institute on Biological Waste Treatment, Manhattan College, New York, N.Y. LAWRENCE, A.W. and McCarty, P.L. (1970), Unified Basis for Biological Treatment Design and Operation, Journal Sanitary Engineering Division, American Society of Civil Engineers, 96, 757. METCALF and Eddy, Incorporated, (1979) Wastewater Engineering Treatment, Disposal, Reuse, McGraw-Hill Book Company, Singapore, 960 pp.


Substrate Concentration, (S) Figure 1: Rate of Substrate Removal in Biological Wastewater Treatment.


Figure 2: Illustrations of Zero Order (Linear) Substrate Removal from Eckenfelder, (1980)


Effluent BOD Concentration

Mean Cell Residence Time, c

Figure 3: Relationship between mean Cell Residence Time and Biological Wastewater Treatment Plant Performance




Wastewater Flows Through All Three Chambers


Wastewater Flows Through Sedimentation Chambers Only


Figure 1: Early anaerobic treatment systems which combined sedimentation and digestion in a single unit





Figure 4: Conventional and high rate separate digestion systems, commonly used for sewage sludge digestion





UPFLOW ANAEROBIC SLUDGE BUCKET Figure 5: Suspended-growth digesters designed to maintain high bacterial populations, allowing digestion at shortened hydraulic detention times




External Control Feed Rate a) Volume b) Solids

Internal Control 2.1 Temperature: a) Mesophilic b) Thermophilic 2.2 (29 370C) (40 600C)

Volatile Acids/Alkalinity Ratio (0.35) An increase in the ratio is the first warning that trouble is starting. Because of the alkalinity in the digester, the pH changes very slowly. In fact, the digester may be completely upset before the pH changes. Frequent monitoring of volatile acid and alkalinity is necessary.











Figure 1: The anaerobic degradation of organic material can be divided into three steps



1. Relationship Of Volatile Acids To Alkalinity


This graph shows a digester operating with a good buffering capacity (the low volatile acids 200 mg/l compared to an alkalinity of 2000 mg/l. At point A, something has happened to cause the volatile acids to increase followed by a decrease in alkalinity at point D. At point G, the digester has become sour.

2. Volatile Acids/ Alkalinity Ratio This graph continues the same digester performance by showing the volatile acids/ alkalinity ratio. Notice that at points CD, the increase in volatile acids produces an increase in the ratio from 0.1 to 0.3.

3. Relationship Of The Change In Ratio Of CO2 To Methane (CH4) As A Result Of 1 By comparing this graph with Graph 2, methane production begins to drop with a corresponding increase in CO2 when the ratio in Graph 2 reaches about 0.5

4. Relationship Of pH Change To Change In 1 pH does not change in this graph until the digester is becoming sour at Point G

Figure 4.9: Graph of change sequence in a digester