50
3 Materials and Methods
Table 3.1 List of instruments
used in the laboratory
Name of equipment
Make and model
Double beam UV-VIS
Spectrophotometer
Systronics
COD digestion apparatus
(Thermoreactor)
E-Merck
BOD Incubator
Instrumentation India
DO Meter
Orion
ION Meter
Orion Star A214
Scanning Electron Microscope Hitachi S 3400 N
Hot Air Oven
Instrumentation India,
Desiccators
Tarsons
Weighing M/c
Afcoset
Muffle Furnace
SAI
BOD Shaker Incubator
Modern Instrument
3.8 Origin and Enrichment of Biomass
Biomass used in the experimental reactor was firstly taken from an activated sludge
process unit. Thereafter, it was enriched under acclimation phase using real municipal
wastewater. Biomass sample was taken from both suspended portion and attached
portion after their growth at steady state in the reactor for their isolation, identification
and DNA study.
3.9 Methodology of Bacterial Identification
3.9.1 Isolation of Pure Cultures
3.9.1.1 Sterilization
Distilled water, 10 test tubes with 9 mL of distilled water each, 10 Petri dishes,
nutrient agar, 250-mL conical flask with 99 mL of distilled water are sterilized in
auto-clave under 15 lb/sq. inch pressure.
3.9.1.2 Serial Dilution
At first, 1 g of active sludge sample taken from the reactor was mixed in 99 mL of
sterile distilled water in the sterilized 250-mL conical flask. After cotton plugging,
the flask was shaken for 1 h. Then, sterilized test tubes containing 9 mL of sterile
3 Materials and Methods
Table 3.1 List of instruments
used in the laboratory
Name of equipment
Make and model
Double beam UV-VIS
Spectrophotometer
Systronics
COD digestion apparatus
(Thermoreactor)
E-Merck
BOD Incubator
Instrumentation India
DO Meter
Orion
ION Meter
Orion Star A214
Scanning Electron Microscope Hitachi S 3400 N
Hot Air Oven
Instrumentation India,
Desiccators
Tarsons
Weighing M/c
Afcoset
Muffle Furnace
SAI
BOD Shaker Incubator
Modern Instrument
3.8 Origin and Enrichment of Biomass
Biomass used in the experimental reactor was firstly taken from an activated sludge
process unit. Thereafter, it was enriched under acclimation phase using real municipal
wastewater. Biomass sample was taken from both suspended portion and attached
portion after their growth at steady state in the reactor for their isolation, identification
and DNA study.
3.9 Methodology of Bacterial Identification
3.9.1 Isolation of Pure Cultures
3.9.1.1 Sterilization
Distilled water, 10 test tubes with 9 mL of distilled water each, 10 Petri dishes,
nutrient agar, 250-mL conical flask with 99 mL of distilled water are sterilized in
auto-clave under 15 lb/sq. inch pressure.
3.9.1.2 Serial Dilution
At first, 1 g of active sludge sample taken from the reactor was mixed in 99 mL of
sterile distilled water in the sterilized 250-mL conical flask. After cotton plugging,
the flask was shaken for 1 h. Then, sterilized test tubes containing 9 mL of sterile