CE5170 – PHYSICOCHEMICAL PROCESSES FOR
WATER AND WASTEWATER TREATMENT
Biological Water Quality
Parameters
BIOLOGICAL WATER QUALITY
PARAMETERS
Aquatic organisms range in size and complexity
Species Diversity Index → parameter for lakes and
streams
Pathogens
→ Capable of infecting or transmitting diseases
→ Requires an animal host
→ Bacteria, Viruses, Protozoa and Helminths
Non-Pathogens
→ Pathogen indicators
→ Analysis of pathogens time consuming
THE MICROBIAL WORLD: SIZES OF MICROBES
Bacterium
0.2-3 µm Protozoan Parasite
Cryptosporidium parvum
~5 µm
Virus
20-80 nm Fungi Parasite
(0.02-0.08 µm) Candida albicans
Larger than 2 µm
BIOLOGICAL WATER QUALITY
PARAMETERS
Bacteria
→ Unicellular organisms
→ Bacilli (Rod shaped)
→ Cocci (Spherical)
→ Spirilla (Spiral)
* Gastrointestinal disorders - symptom
* Example - Vibrio comma; Salmonella typhosa, etc.
→Growth conditions and environmental conditions
may be different for different bacteria
→ Example – Psychrophilic (0 to 20oC), Mesophilic (20
to 45oC) and Thermophilic (> 50oC)
Bacteria’s morphology
BIOLOGICAL WATER QUALITY
PARAMETERS
Viruses
Simplest includes nucleic acids (DNA or RNA) and protein capsid
Absolute parasites
Various disease with symptoms of fever, vomiting, jaundice.
Size of viruses between 0.02-0.08 µm
Conc. of viruses in raw wastewater 103-104/100 ml, with infectious
dose 1-10
Penetration of viruses to groundwater is more likely than bacteria
due to their size, and also due to the extreme high numbers in
infected individuals (infected person – 105 – 1011 viruses per gram
of feces)
Viruses feature longer survival time than bacteria and more
resistant to disinfection
http://techalive.mtu.edu/meec/module02/Viruses.htm
http://health.howstuffworks.com/medicine/modern/light-virus.htm
HUMAN ENTERIC VIRUSES
More than 100 types of enteric viruses capable of
causing infection and excreted by humans
Enteric viruses multiply in the intestinal tract and are
released in fecal matter of infected persons
Most important human enteric viruses are: polio,
coxsackie, Norwalk virus, rotavirus, adenovirus and
hepatitis A virus.
The pathogen virus HIV is not waterborne
POLIO VIRUS-POLIOMYELITIS
50 years ago one of the most feared
diseases
Symptoms are like flu, slight fever,
headache, sore throat and up to deep
muscle pain and paralysis
Transmission fecal to oral route
Found in wastewater and water
ROTAVIRUS
Causing severe diarrheal disease in
humans (30-50% of all cases)
80 nm in diameter
Appearance of little wheels viewed
by EM (rota)
dsRNA in a capsid
Symptoms: vomiting, watery
diarrhea, fever
Person with disease can excrete
10^8-10^10 units/ml of feces
Infective dose 10-100 units
ADENOVIRUSES:
~80 NM DIAMETER
• Only human enteric virus
with Double-stranded DNA
• Protein coat with
attachment fibers
• Infection in lungs, eyes,
genitals
• Possible agent in
waterborne outbreaks
(found in water)
BIOLOGICAL WATER QUALITY
PARAMETERS
Cryptosporidium parvum - Protozoa
→ Lowest form of animal life
→ Unicellular (complex functional activity)
* Symptoms: Gastrointestinal disorders
* Example – Giardia Lamblia, Entamoeba histolytica
Exist as a Oocyst (3-6 µm) or Sporozoite
Infectious dose 1-10 – no effective treatment
GIARDIA LAMBLIA-SINGLE CELLED, PROTOZOA
Giardiasis - symptoms of mild to
severe diarrhea, nausea and
indigestion
Exist as a Cyst or Trophozoite.
The cyst is the environmentally
resistant form ( in water)
Size: µm 6-8 W × 8-14 L
Infectious dose < 20
BIOLOGICAL WATER QUALITY
PARAMETERS
Helminths
→ Parasitic worms
→ Contaminated by aquatic species (snails, insects)
→ Easy to destruct by disinfection
❑ Water borne pathogens → Introduced through fecal
contamination
❑ Intestinal tract organism are good indicator organism
❑ Fecal Coliform group (FC) → for wastewater quality
❑ Total Coliform group (TC) → for water quality
SERIAL DILUTION
IDENTIFICATION OF BACTERIA
Direct count
Microscopically or Electronically (based on EC)
→ Differentiation of live and dead cells not possible
→ Stain can be to count different bacteria
Pour plate method
→ Dilute sample serially
→ Diluted sample + agar medium →then solidify
→ Incubate and count the cells as cfu/mL
Spread plate method
→ Spread sample on top of the solid medium
IDENTIFICATION OF BACTERIA
Membrane filtration technique
Filter sample using 0.45 µm
Remove the filter paper with solids (microbes)
Incubate and count the colonies
TESTING FOR COLIFORM BACTERIA:
MEMBRANE FILTRATION
Membrane filter
0.45 μm pores
47 mm in diameter
Filter 100 mL of water to
be tested through the
membrane filter
MEMBRANE FILTRATION
Add 2 mL of m- Place membrane
Petri dish with endo broth filter in the petri
sterile absorbent (selective dish on top of
nutrient pad media) the nutrient pad
MEMBRANE FILTRATION:
INCUBATION AND RESULTS
Incubate for 24 hours at
35°C 2
1
Coliform bacteria grow
4
into colonies with a 3
green metallic sheen 5 6
Non-coliform bacteria
8
may grow into red 7
colonies
Coliform concentration is
__________________
8 coliform/100 mL
MULTIPLE TUBE FERMENTATION
TECHNIQUE
Coliform bacteria → Ferment lactose
Test has three stages
→ 1. Presumptive test
→ 2. Confirmed test
→ 3. Completed test
Gas formation is considered as the +ve indication
From the test results →estimate the MPN
MPN → Most probable number
MPN determined by: (1) Poisson distribution, (2)
MPN Tables and (3) Thomas equation
MULTIPLE TUBE FERMENTATION
TECHNIQUE
3
MOST PROBABLE NUMBER
Poisson distribution
Where,
y → Probability of occurrence of a given result
a → constant for a given set of conditions
n1, n2, n3 → sample size in each dilution
λ → Coliform density, number/mL
p1, p2, p3 → number of positive tubes in each sample dilution
q1, q2, q3 → number of negative tubes in each sample dilution
Thomas Equation
MOST PROBABLE NUMBER
The result of a coliform analysis using the
multiple-tube fermentation test of the effluent
from an intermittent sand filter are given below.
Using these data, determine the coliform density
(MPN/100 mL) using the Poisson equation, the
Thomas equation and the MPN Table.
Size of portion, Number Number
mL positive negative
1.0 4 1
0.1 3 2
0.01 2 3
0.001 0 5
λ Ya (x10-7)
SOLUTION 3.80
3.84
3.6754
3.6773
Poisson Distribution → 3.85 3.6774
3.86 3.6773
MPN/100 mL = 100 x λ
3.90 3.6755
= 385/100 mL
Thomas Equation
No. of positive tubes =9
mL of sample in –ve tubes = 1.235 mL
mL of sample in all tubes = 5.555 mL
→ MPN/100 mL = 344/100 mL
MPN Table
→ 4-3-2 ➔ 39 x 10 = 390/100 mL
