1- Microbiology

Bacteria vary widely in their environmental requirements and preferences. For example,
some bacteria can only grow in the presence of oxygen - the 'obligate aerobes'; some
can only grow in its absence - the 'obligate anaerobes'; and some can grow in both its
presence and in its absence, although growth is better in its presence - the 'facultative
anaerobes' (or simply 'facultative bacteria').

Most bacteria cannot use carbon dioxide as a source of cell carbon, but some can (and
prefer to do so); those that cannot are termed 'heterotrophs' and those that can are
'autotrophs'.

Some bacteria can photosynthesize, and these are either 'photoheterotrophs' or 'photo-
autotrophs'; however, most bacteria cannot photosynthesize and they are therefore either
'chemoheterotrophs' or 'chemoautotrophs'.

Temperature is a very important environmental parameter. Most bacteria grow well in the
temperature range 15 - 40 C and are termed 'mesophils'; some grow best at lower
temperatures - the 'psychrophils'; and some require much higher temperatures (some
even close to the boiling point of water) - the 'thermophils'. In wastewaters in tropical
and subtropical regions most bacteria are, as would be expected, mesophilic.

The pH of the environment in which bacteria grow is another important environmental

parameter. Most bacteria prefer near neutral or slightly alkaline conditions, around pH
6.5 - 8.5; some can tolerate pH > 9 (e.g. Vibrio cholerae, the causative agent of cholera);
and some generate very acid conditions (e.g. Thiobacillus thioparus which produces
sulphuric acid at pH < 2 and so causes rapid sewer crown corrosion in warm climates).

Salt is an environmental parameter of importance, for wastewater treatment engineers,

only in that freshwater and faecal bacteria cannot grow in very saline waters (the sea, for
example; this is relevant when treated wastewaters are discharged into coastal waters).
Marine bacteria, in contrast, are 'halophils'.

Domestic wastewater fortunately contains roughly the right balance of nutrients for
bacterial growth - a BOD:N:P ratio of ~100:5:1. The presence of industrial effluents can
alter this ratio and the wastewater may need nitrogen and/or phosphorus supplements.

Anabolism and catabolism

Bacteria oxidize wastes to provide themselves with sufficient energy to enable them to
synthesize the complex molecules such as proteins and polysaccharides which are
needed to build new cells. Thus bacterial metabolism has two component parts:
catabolism ('breaking down') for energy and anabolism ('building up') for synthesis. The
verbal 'equation'
is instructive but oversimplified in that the anabolic and catabolic reactions are not
distinguished; nor is there mention of autolysis, which is an important form of
catabolism. The following three equations describe these processes separately:
 Catabolism

 Anabolism

 Autolisis

As a general guide ~1/3 of the available BOD is used in catabolic reactions and ~2/3 in
anabolic reactions (Figure below). The equation for autolysis does not proceed to
completion since approximately 20-25 per cent of the cell mass is resistant to bacterial
degradation.

Note: In a real (finite time) continuous microbiological reactor some of the organic
matter (i.e. BOD) in the influent escapes oxidation; in batch culture at infinite time
the unmetabolized fraction is zero.

Figure The Catabolic, Anabolic and Autolytic Reactions of Aerobic Microbiological

Oxidation
 2- Kinetics of Biological Growth

2-1) Logarithmic Growth:

dX
rg  X  (1)
dt

where rg is rate of cell growth (mass/unit volume . time), X is concentration of micro-

organisms (mass / unit volume) and  is specific growth rate (time-1).

2-2) Substrate Limited Growth:

S
  m (2)
Ks  S

where S is concentration of substrate (mass / unit volume) and  m is maximum value of

specific growth rate (time-1). At high concentrations of substrate ( S  K s ) specific
growth rate is maximum (   m ) and specific growth rate is equal half of its maximum
value (   m 2 ) when S  K s .

/m 1

m/2

0
Ks S

If the value of  from Eq. 2 is substituted in Eq. 1, the resulting expression for the rate of
growth is
  S 
rg    m X (3)
 Ks  S 

2-3) Cell Growth and Substrate Utilization:

It has been observed that for a given substrate the quantity of new cells produced is
reproducible. Maximum yield coefficient, the ratio of the mass of cells formed to the
mass of substrate consumed (mass/mass) over any finite period of logarithmic growth,
can be used to quantify the relation between rate of cell growth and rate of substrate
utilization as

rg   Yrsu (4)

where rsu is substrate utilization rate (mass/unit volume . time). Solving Eq. 4 for rsu and
substitution of rg from Eq. 3 leads to

m  S   S 
rsu     X  k  X (5)
Y  Ks  S   Ks  S 

where k  m Y is defined as maximum rate of substrate utilization per unit mass of

micro-organisms.

1-4) Effect of Endogenous Metabolism:

Rate of cell death due to endogenous respiration, rd , is an essential factor that should be
included in any rate equation developed to describe the rate of cell growth.

rd  k d X (6)

where k d is endogenous decay coefficient (time-1).

1-5) Net Rate of Biological Growth:

Eqs. 3 and 6 present the rates of new cell production due to substrate utilization and cell
decay due to endogenous respiration regime. A rate equation that includes these two rate
equations can be considered as the net rate of biological growth, rg

rg  rg  rd   Yrsu  k d X (7a)

 S   S 
rg   m  X  k d X   m  k d  X   X (7b)
 Ks  S   Ks  S 

where  is the net specific growth rate (time-1). The relation between net rate of
biological growth and rate of substrate utilization can be expressed as
 rg
Yobs   (8)
rsu
where Yobs is defined as observed yield and has the same units as maximum yield
coefficient, Y .

3- Application of Kinetics to Biological Treatment

The application of biological growth kinetics for developing operation equations for a
CSTR is discussed here. The operation equations are derived from micro-organisms and
substrate balances and can be used to (a) predict the concentration of micro-organisms
and substrate in CSTR effluent, (b) develop process design factors and (c) examine the
effect of kinetics on process design factors and performance.

Q, S0, X0 Q, S, X

Reactor

3-1) Micro-organism Mass Balance:

Accumulation  inf low  outflow  net growth (9)

V
dX
dt
 QX0  QX  V rg   (10)

where V is reactor volume. Substitution of rg from Eq. 7b leads to

dX  S 
V  QX0  QX  V  m  kd  X (11)
dt  Ks  S 

Steady state solution of Eq. 11 under the reasonable assumption of almost zero inlet
concentration of micro-organisms, X0  0 , is required.
   S 
Q  V   m  kd  X  0 (12)
  Ks  S  

The fact that the concentration of micro-organisms in effluent cannot be zero, X  0 ,

leads to the first operating equation (put the coefficient of X equal to zero and
rearrange).

Q 1 S
  m  kd (13)
V  Ks  S

where  denotes hydraulic retention time. Eq. 13 can be rearranged as

S 1 1 
   kd  (14)
K s  S m   

3-2) Substrate Mass Balance:

Substrate mass balance is as follows

dS
V  QS0  QS  V  rsu  (15)
dt

Substitution of rsu from Eq. 5 leads to

dS  S 
V  QS0  QS  V  k X (16)
dt  K s  S 

The second operating equation can be derived by rearranging the steady state solution of
Eq. 16, dS dt  0 as

S
S0  S  k X0 (17)
Ks  S

3-3) Predictions of Effluent Micro-organisms and Substrate

Concentration:
All is required to derive an equation to predict micro-organisms concentration, X , is
substitution for S  K s  S  from Eq. 14 in Eq. 17 and solving it for X as

m  S0  S  Y  S0  S 
X  (18)
k  1  k d  1  k d
To derive an equation that allows prediction of substrate concentration, S , one need to
(a) solve Eq. 17 for S  K s  S  as

S S S
 0 (19)
Ks  S kX

(b) substitute for X in Eq. 19 from Eq. 18 as

S S0  S
 (20)
Ks  S  Y  S0  S  
k  
 1  k d 

and (c) solve Eq. 20 for S and rearrange as

K s  1  k d 
S (21)
  Yk  k d   1

For a given process, having the values of the four process parameters K s , k d , Y and k
allows one to use Eq. 21 to calculate effluent substrate concentration, S and subsequently
Eq.18 to calculate micro-organisms concentration, X .

4- Determination of Kinetic Coefficient

To determine the value of the four process parameters K s , k d , Y and k , bench-scale

aerobic and anaerobic are used. The usual procedure is to operate the reactor over a range
of effluent substrate concentration. This can be achieved by selecting several different
values for hydraulic retention time,  , within the range of 1 to 10 days. Each
experimental run should be continued until steady state conditions are reached. Mean
values of the collected data under steady state conditions for Q , S 0 , S and X are required
to calculate the four process parameters by using the following two equations.

X K 1 1
 s   (22)
S0  S  k  S k

1 r
  Y su  k d (23)
 X
In the first step, values of K s and k can be determined by plotting X  S0  S 
versus 1 S . The values of k d and Y can then be determined by plotting of 1 
versus  rsu X   kS  K s  S  .

5- Aerobic Suspended-Growth Treatment

These processes include a) Activated-Sludge, b) Suspended-Growth Nitrification,
c) Aerated Lagoons, d) Aerobic Digestion Process and e) High-Rate Oxidation Ponds.
Among these activated sludge is the most important one.

5-1) Activated Sludge:

This process was first developed in England in 1914 by Ardern and Lockett. Its naming is
due to the fact that in activated sludge process involves production of an activated mass
of micro-organisms that are capable of aerobically stabilizing waste. The schematic
diagram of activated sludge process is shown below.

Qw, X

Reactor Settling Tank

Q, S0 (Q-Qw), S, Xe

X, V, S

Qr, S, Xr

In this diagram the subscripts w, r and e represent properties of waste, recycle and
effluent streams. The reactor is a CSTR that is kept well aerated by mechanical or
diffused aerators and it is assumed that waste stabilization occurs only in reactor. The
settling tank allows separation of active biomass in sludge phase from clear-treated
effluent stream. The recycle stream returns the active biomass to the reactor so that waste
stabilization can be performed effectively. To avoid accumulation of biomass in the
reactor the waste stream, on top of the reactor, carries away some biomass as waste.

Vs
Mean Hydraulic Retention Time for the System, s  (24)
Q

where Vs denotes volume of the system, volume of reactor plus volume of settling tank.
 V
Mean Hydraulic Retention Time for the Reactor,   (25)
Q

VX
Mean Cell Residence Time, c  (26)
Q w X   Q  Q w  Xe
Under the assumption of effective settling unit, Xe is very small, the mean cell residence
time is

V
c  (27)
Qw

The operating equation of this system can be calculated by writing a micro-organism

mass balance as

V
dX
dt
 
 QX0  Qw X   Q  Qw  Xe   V rg (28)

The steady state version of Eq. 28 can be derived by considering  dX dt   0 ,

substituting for rg from Eq. 7a and rearranging.

Qw X   Q  Qw  Xe  r
  Y su  k d (29)
VX X

Eq. 26 allows for replacement of the left hand side of Eq. 30 with 1 c .

1 r
  Y su  k d (30)
c X
 6- Aerobic Attached-Growth Treatment
These processes include a) Trickling filter, b) Roughing filter, c) Rotating biological
Contactor and d) Fixed-bed Nitrification reactor. The trickling filter is used most
commonly and is discussed here in detail.

6-1) Trickling Filter:

The first trickling filter designed and operated in 1893 in England. This process can be
considered as the continuous version of the batch contact beds in which a fixed bed of
broken stones was (a) filled with wastewater from top, (b) allowed to be in contact with
wastewater for a short time and (c) drained and rested before repeating another cycle. The
long duration of each cycle of contact beds, 12 hours, and other limitations of contact
beds is eliminated in trickling filters. Modern trickling filters usually consist of shallow
basin with circular cross section that is filled with filter media. The filter media is usually
rocks with sizes varying from 25 to 100 (mm). The depth of filter media is in the range of
0.9 to 2.5 (m). Trickling filters with plastic packing have a filter media depth of 9 to 12
(m) and usually are built with square cross section. The following schematic diagram
presents involving phases in the operation of trickling filter and direction of mass transfer
of different chemical.
Liquid Air
waste

Organics
Media

Biological O2
mass
CO2

End
products

This next diagram is presented in a format that helps for developing a mathematical
model describing operation of trickling filter.
Q, S

w
Z

Slime layer rs
Liquid layer
Z+dZ
h 

Q, S+[(S/Z)dZ]
Under the assumptions of (a) the process of waste stabilization is controlled by diffusion
of organic matter in slime layer and (b) there is no concentration gradient across liquid
film the following equation is proposed to describe the rate of flux of organic matter into
slime layer.

Ehk 0S
rs   (31)
Km  S

where
rs = rate of flux of organic matter into slime layer
E = effectiveness factor ( 0  E  1 )
h = thickness of slime layer
k0 = maximum reaction rate
S = average BOD in the bulk liquid of the element
Km = half-velocity constant

It is proved that effectiveness factor is proportional to BOD

E  BOD or E  fS (32)

Substitution of E in Eq. 31 from Eq. 32 leads to a new rate equation as

fhk 0S 2
rs   (33)
Km  S

Operating equation of a trickling filter can be derived by using Eq. 33 in a mass balance
of organic matter over the liquid film as

S  S   fhk 0S 
2
dV  QS  Q  S  dZ      dZw (34)
t  Z   Km  S 

where w and dZ are the width and height of the liquid element. In most practical cases,
S  K m . This allows one to simply the steady state version of Eq. 34 to

dS wS
 fhk 0 (35)
dZ Q

The variables of Eq. 35 can be separated and integrated as

Se Z
dS w
S S  0 fhk 0 Q dZ (36)
i
 Se  wZ 
 exp    fhk 0  (37)
Si  Q 

where S i and S e are the BOD of wastewater applied to and the effluent stream from
filter. Application of Eq. 37 requires steady state operation of the filter over a wide range
of variable group of wZ Q and collecting data for S i and S e . In a plot of  ln  Se Si 
versus wZ Q , the slope of the best line that can be passed through the experimental data
and forced to pass through the origin provides the value of fhk 0 .