INTRODUCTION TO BIOLOGICAL WASTEWATER TREATMENT

The basis of biological wastewater treatment is the use of bacterial population to convert soluble
organic matter in wastewater into insoluble organic material (more bacteria) which then can be
physically removed from the wastewater stream. The result is the elimination of oxygen
demanding organic chemicals from the wastewater, this permits discharge or reuse of the clean
water.

1) THE BACTERIA – BIOLOGICAL GROWTH THEORY

The primary types of bacteria involved in the biochemical oxidation of organic compounds are:

Bacillus - rod shaped organisms. Approximately 2 – 5 microns in

length and 1 – 2 microns in diameter.

Coccus - spherical shaped organisms. Approximately 1 – 2 microns

in diameter.

Spirochetes - spiral shaped organisms.

For developing a simplified model of bacteria and how they function, we will use bacillus as an
example. The bacillus cell is composed essentially of proteinaceous material. Its primary
directive is to survive and reproduce. In biological wastewater treatment systems the
reproduction or growth of bacterial populations results in a conversion of undesirable soluble
organic materials present in wastewater to cell material or protein. Thus, the soluble organic
chemicals are removed from the wastewater.

The growth of bacteria involves a number of functions, but most critical is the oxidation of a
carbon source (the organic chemical) through respiration (use of oxygen) which generates energy
(plus carbon dioxide and water) that can be utilized for cell maintenance, metabolism and
reproduction of additional cells.
The following are definitions of the terms useful in understanding the operation of wastewater
systems.

SOLUBLE ORGANIC MATTER – These are the organic chemicals dissolved in

wastewater which we wish to remove from the water and which the bacteria seeds as a
source of food or carbon.

EXOENZYMES – These are biochemical compounds released by bacteria into their

environment. The exoenzymes facilitate the conversion of solid organic materials (oils,
grease, fats) into soluble organic matter.

NUTRIENTS – In order for the bacteria to perform their function, certain essential
nutrients are required such that the metabolic processes can occur and additional cellular
material may be built. The most important of these are:

NH3-N ammonia nitrogen

PO4-P soluble orthophosphate
Trace metals Iron, Zinc, Cobalt, etc.

Inadequate amounts of these essential nutrients will substantially retard the growth rates
of these organisms and, hence, their ability to metabolize and remove the organic material
from the wastewaters.

BIOPOLYMERS – These substances are composed of polysaccharides which are secreted

by the organisms and coat the outer cell wall. These biopolymers are essential to the
biological wastewater treatment process in that they allow the agglomeration of numerous
organisms into floc particles or slimes. Growth of the bacteria as a slime formation on
surfaces is critical to operation of biological treatment systems such as the CTX-BIOX
systems. Formation of floc permits the use of gravity separation in biological systems
such as activated sludge systems. Formation of slimes and floc by bacteria thus provides
a means of separating the bacteria from wastewater and permits a clarified effluent to be
discharged from the biosystem.

METABOLIC BY-PRODUCTS – These are by-products excreted by microorganisms.

They are composed of intermediate chemical breakdown products, CO2 and H2O. Other
bacterial populations will utilize the intermediate breakdown products as their food
source and subsequently excrete other breakdown products. Eventually, the oxidation of
the undesired organic compound is completed.

MORE BACTERIA – These are the reproductive offspring of the original bacteria. In
essence it is a mass of protein (bacterial cells) which has been produced from the
undesirable organic chemicals by the bacteria.

The organisms assimilate and metabolize their organic food source as they grow and reproduce.
The reproduction of the organisms occurs, for instance, by cell division with every parent cell

-2-
dividing into two cells. The time period for this division to take place in organic carbon
consuming bacteria under ideal conditions is a minimum of 15 minutes. It will be much slower
in actuality due to practical limitations in the use of food nutrients and oxygen.

If an average doubling rate or cell generation time of 20 minutes is assumed for the bacteria and
we start with one cell per liter after 10 cell generations (200 minutes) we would have more than
1,000 bacterial cells. This increase is logarithmic and billions of cells are generated very rapidly.
As the concentration of available food for the organisms increases, the rate at which the
organisms grow increases dramatically until it reaches a maximum rate. The Monod curve
describes these phenomena.

Based on this curve, the growth rate of the organisms increases dramatically with an increase in
available food (substrate) concentration and similarly slows down as the concentration decreases.
This is the dynamic response of microbial populations to the changes in food concentration. The
collective population in a biological wastewater system exhibits the same response
characteristics. As the loading (i.e., lbs./day input) of organic chemicals on the wastewater
treatment plant increases, the growth rate of the organisms increases. The additional organics are
metabolized and additional bacteria are produced. If the loadings drop below that required to
maintain the existing number of bacteria, there will be a decline in the concentration of
organisms in the biological system. Essentially, the organisms are being starved and they will die
off until their numbers are reduced to a level at which the food input to the system is sufficient to
maintain them.

We can see that certain nutrient requirements are the key to successful biological growth and
conversion of the soluble organic to the insoluble (bacterial) form. It is also important to
remember that bacteria cannot survive without the appropriate working environment.

ENVIRONMENTAL FACTORS

In addition to the nutrient and food requirements discussed above, certain environmental
conditions are requisite for biological growth.

pH

Appropriate pH conditions for biological growth generally are between 6 – 9 pH units. The
ability of the organism to survive and function on either side of this range is marginal. The
excessive concentration of either H+ or OH- in the microbe’s environment can inhibit metabolic
activity and can also cause physical breakdown of the organism itself. Optimum pH for
biotreater operation is usually 6.5 – 7.5.

TEMPERATURE

This condition will greatly affect the rates at which the microbes function. The rate at which
bacteria can utilize the organic chemicals in wastewater is directly related to the water
temperature in which they function. For every 10 C increase in water temperature, the rate of

-3-
metabolic activity will double. Inversely, as the temperature drops 10 C the metabolic activity
will be cut in half. For example, at 10 C the rate of metabolic activity will be approaching zero,
since the bacteria cannot function effectively at this temperature. At 40 C, physical
characteristics of the collective biological populations begin to change and they are difficult to
keep in the biological wastewater treatment system. At some point above 45 C - 50 C the
enzymes which are required to facilitate breakdown to the organic food source will begin to
break down due to thermal conditions and the bacteria die. Optimum treatment temperatures are
generally between 20 C and 30 C.

H2O

Water is a necessary medium for survival of the microorganism. It is essential for transport of
materials in and out of the cell as well as within the cell.

< 60 F Activity slows

< 45 F No growth
> 30 F Kills bacteria
> 90 F Destroys enzymes – inhibits bacteria
> 105 F Kills bacteria

Dissolved Oxygen 2 mg/l – 7 mg/l

< 2 mg/l oxygen limited

< 0.5 mg/l anaerobic
> 7 mg/l slow growth or overdesign
> 9 mg/l bacteria dead

Nutrient balance

BOD : N : P 100 : 10 : 2
(keep residual of 1 – 2 ppm of NH3 – N and PO4 – (P in effluent)

Operation Limits

pH 6 9

< 6.5 Nitrification stops

< 6.0 Activity slows
< 5.0 Bacteria start to die
< 4.0 Pickled bacteria
> 9.0 Activity slows

NH3 Soluble effluent 1 mg/l

-4-
 (except when nitrification is occurring)
< 1 mg/l slow response to shocks
< 0.2 mg/l inhibit growth

-5-
 BIOLOGICAL SYSTEMS

ACTIVATED SLUDGE

GENERAL INFORMATION

1) BACTERIA SPLIT INTO IDENTICAL CELLS

A. Occurs every 20 to 30 minutes
B. 24 hours bacteria equal to Earth’s volume
C. Increased waste removal – food, oxygen & nutrients
D. Treatment plants supply adequate oxygen and nutrients
E. Food supply is limiting factor and affects growth rate

2) GROWTH/DEATH OF BACTERIA – FOUR PHASES

A. Phase I - Increasing growth rate – rapid bacterial production
B. Phase II - Stationary – growth rate equal to death rate
C. Phase III - Decreasing death phase
D. Phase IV - Endogenous – nutrients from dying/lysed bacteria provide
other bacteria with food

3) TREATMENT TIME
A. Residence time – bacteria reduce incoming, organic load BOD down to levels
below NPDES permit
B. Phase I to Phase II – young bacteria – high sludge production – sludge not
stabilized / water demand high / aerobic odors
C. Phase III & IV – BOD removal high/sludge low – high economic to
maintain
D. Sludge plants – maintain between Phase II & III

4) TREATMENT PLANT – FACTORS – BESIDES ECONOMICS

A. Remove BOD and minimize solids
B. Settle-ability of solids/bacteria enter death phase, they secrete natural
polymers and enzymes forming a gelatinous matrix around cell walls. Particles
settle well.
C. Phase I and Phase IV floc is dispersed

5) RETENTION TIME: (RESIDENCE TIME OF BACTERIA)

A. Basin size
B. Concentration of bacteria (MLSS)
C. Aeration system (multiple)
D. Sludge return to aeration basin or clarifier

-6-
REQUIRED CONTAINERS, PRESERVATION TECHNIQUES, AND HOLDING TIMES

MAXIMUM
NAME CONTAINER PRESERVATION HOLDING
TIME
Bacterial Tests:
Coliform, fecal and total P, G Cool, 4C, 0.008% Na2S2O3 6 hours
Fecal streptococci P, G Cool, 4C, 0.008% Na2S2O3 6 hours
Inorganic Tests:
Acidity P, G Cool, 4C 14 days
Alkalinity P, G Cool, 4C 14 days
Ammonia P, G Cool, 4C, H2SO4 to pH<2 28 days
Biochemical oxygen demand P, G Cool, 4C 48 hours
Bromide P, G None required 28 days
Biochemical oxygen demand,
carbonaceous P, G Cool, 4C 48 hours
Chemical oxygen demand P, G Cool, 4C, H2SO4 to pH<2 28 days
Chloride P, G None required 28 days
Chlorine, total residual P, G None required Analyze immediately
Color P, G Cool, 4C 48 hours
Cyanide, total & amenable to Cool, 4C, NaOH to pH>12,
Chlorination P, G 0.6g ascorbic acid 14 days
Fluoride P None required 28 days
Hardness P, G HNO3 to pH<2, H2SO4 to pH<2 6 months
Hydrogen ion (pH) P, G None required Analyze immediately
Kjeldahl and organic nitrogen P, G Cool, 4C, H2SO4 to pH<2 28 days
Metals:
Chromium VI P, G Cool, 4C 24 hours
Mercury P, G HNO3 to pH<2 28 days
Metals, except chromium VI
and mercury P, G HNO3 to pH<2 6 months
Nitrate P, G Cool, 4C 48 hours
Nitrate-nitrite P, G Cool, 4C, H2SO4 to pH<2 28 days
Nitrite P, G Cool, 4C 48 hours
Oil and grease G Cool, 4C, H2SO4 to pH<2 28 days
Organic carbon P, G Cool, 4C, HCL or H2SO4 to pH<2 28 days
Orthophosphate P, G Filter immediately, Cool, 4C 48 hours
Oxygen, Dissolved Probe G Bottle & top None required Analyze immediately
Winkler Do Fix on site & store in dark 8 hours
Phenols G only Cool, 4C, H2SO4 to pH<2 28 days
Phosphorus (elemental) G Cool, 4C 48 hours
Phosphorus, total P, G Cool, 4C, H2SO4 to pH<2 28 days
Residue, total P, G Cool, 4C 7 days
Residue, Filterable P, G Cool, 4C 7 days
Residue, Nonfilterable (TSS) P, G Cool, 4C 7 days
Residue, Settleable P, G Cool, 4C 48 hours
Residue, Volatile P, G Cool, 4C 7 days
Silica P Cool, 4C 28 days
Specific conductance P, G Cool, 4C 28 days

-7-
 CONCENTRATION OF POLLUTANTS THAT MAKE PREBIOLOGICAL OR
PRIMARY TREATMENT ADVISABLE

Pollutant or System Condition Limiting Concentration Kind of Treatment

Sedimentation, flotation, or
Suspended solids >50 to 125 mg/liter lagooning
Oil or grease >35 to 50 mg/liter Skimming tank or separator
Toxic ions Precipitation or ion exchange
Pb  0.1 mg/liter
Cu + NI + CN  1 mg/liter
Cr-6 + Zn  3 mg/liter
CR+3  10 mg/liter
pH <6>9 Neutralization
0.5 lb. alkalinity as Neutralization for excessive
Alkalinity CaCO3/lb. BOD removed alkalinity
Acidity Free mineral acidity Neutralization
Organic load variation > 2:1 to 4:1 Equalization
Precipitation or stripping with
Sulfides > 100 mg/liter recovery
Extraction, adsorption, or
Phenols > 70 to 300 mg/liter internal dilution
Dilution, ion exchange, pH
Ammonia > 1.6 g/liter adjustment or stripping
Dissolved salts > 10 to 16 g/liter Dilution or ion exchange
Temperature 13 to 38 C in reactor Cooling or steam addition

-8-
 HEAVY METALS
METAL, CYANIDE & INORGANIC COMPOUND CONCENTRATIONS
INHIBITING BIOLOGICAL PROCESSES (IN MG/L)

Biological Process
Activated Nitrification Aerobic Anaerobic
Pollutant Sludge Fixed Film Digestion
Ammonia <480 N/A N/A 1,500 – 3,000
Arsenic 0.04 – 0.4 N/A 290 0.1 – 1
Boron 0.05 – 10 N/A N/A 2
Cadmium 0.5 - 10 5–9 5 – 20 0.02 – 1
Calcium 2,500 N/A N/A N/A
Chloride N/A 180 N/A 20,000
Chromium (Tot.) 0.1 – 20 0.25 – 1 50 1.5 – 50
Copper 0.1 – 1 0.05 0 0.5 25 – 50 0.5 – 100
Cyanide 0.05 – 20 0.3 – 20 N/A 0.10 – 4
Iodine 10 N/A N/A N/A
Iron 5 – 500 N/A N/A 5
Lead 0.1 – 10 0.5 – 1.7 N/A 50 – 250
Manganese 10 N/A N/A N/A
Magnesium N/A 50 N/A 1,000
Mercury 0.1 – 5.0 2 – 12.5 N/A 1,400
Nickel 1–5 0.25 – 5 N/A 2 – 200
Silver 0.03 – 5 0.25 N/A N/A
Sodium N/A N/A N/A 3,500
Sulfide >50 N/A N/A 50 – 100
Tin N/A N/A N/A 9
Vanadium 20 N/A N/A N/A
Zinc 0.30 – 20 0.01 – 1 N/A 1 – 10

N/A = Not Available

-9-
 ALWAYS ASK!
1. What is the problem and cause?
2. What type of biological process does the treatment plant contain?
3. What is the size and volume of the process?
4. What units are there prior to and following biological process?
5. What are the sizes and volumes of the processes?
6. What is the Flow (Q) and is it continuous?
7. What is the Retention time in biological process?
8. What is the Food to Mass ratio (F/M)?
9. Does the plant get any toxic shocks?
10. What are the toxic shocks, the concentrations, and frequency?
11. What type of aerators:
Horsepower (HP)
Number
12. What is the Temperature (C or F) at the influent (inf) and effluent (eff)?
13. What is the pH of the influent, effluent and the permit limits?
14. What are the Mixed Liquor Suspended Solids (MLSS)?
15. What is the Biochemical Oxygen Demand (BOD) of the influent, effluent and the
permit limits?
16. What is the Chemical Oxygen Demand (COD) of the influent, effluent and the permit
limits?
17. What is the Alkalinity (Alk) of the influent, effluent and the permit limits?
18. What are the Total Suspended Solids of the influent, effluent and the permit limits?
19. What is the Ammonia Nitrogen (NH3N) or Total Kjeldahl Nitrogen (TKN) of the
influent, effluent and the permit limits?
20. What is the Orthophosphate (O-PO4) of the influent, effluent and the permit limits?
21. Is there any return to the biological processes and Rate of Return (RAS)?
22. Any acids or caustics used in plant, what are the concentrations and time period of
use?
23. What Chemical additives are used? i.e., Lime, Ferric, Alum, Polymer?
24. What is the Dissolved Oxygen (D.O.) in the biological process?
25. Are there any Metals in the waste stream and what are the levels?
26. What is the Oil and Grease Level of the influent, effluent and the permit limits?
27. Are there any other permit levels that must be reached?
28. Is the treatment facility currently using biological products?
29. What products are they using, their cost and the dosage?

- 10 -
 ALWAYS ASK!
FOR INDUSTRIAL PLANT:

1. Is the flow continuous?

2. What are the type of plant operations and time of operations?
3. Are there any Wash Down Periods and what is used?

FOR AMMONIA PROBLEMS:

1. What caused Ammonia upset?

2. Has sludge been wasted recently?
3. Has chlorine been used to achieve ammonia reduction?
4. What is the current percent of ammonia removal?
5. Is it an ammonia problem and not an amine problem?

FOR FILAMENTOUS PROBLEMS:

1. What type of filamentous growth is it?

2. What color is it?
3. What is its texture?
4. What is the amount of foam, its size and depth?
5. How long is the sludge retention time in the clarifiers?
6. Is there floating sludge in the clarifiers?
7. Do you chlorinate or use Hydrogen Peroxide (H2O2)?
8. Have there been any spills or upsets in operations?
9. Have there been any plant shut downs?

FOR PHENOLS:

1. What type of phenols are used.

2. What are the influent, effluent levels and permit levels?

FOR HYDROCARBON OIL AND GREASE:

1. What type of hydrocarbons are to be removed?

2. Are they floating on basin or clarifier?
3. What are the level of hydrocarbons and the permit limits?

- 11 -