Microbiology @ IHS Laboratory (BioLab).
The BioLab is equipped to analyse and test for microbial contamination of water. Various plate count tests are available to estimate bacterial colony forming units (cfu), which is a measure of the total bacterial population in a water sample. Multiple tube dilution technique is used to estimate most probable number (MPN) of total coliforms, some of which would indicate environmental and faecal contamination of water. Confirmatory tests are available to identify indicator organisms associated with environmental and sewage contamination. The bacterial endotoxin assay is available to test the level pyrogens (endotoxins) in water for dialysis.
The plate count methods rely on growth of bacterial colonies on a variety of nutrient mediums incubated at specific temperature for various durations, so that colony becomes visible to the naked eye. But too many colonies, some of which may coalesce with each other would be difficult to count. Hence multiple petridishes with chosen nutrient medium are inoculated with different dilutions of test sample and the colony count is multiplied by the dilution factor.
In the most probable number (MPN) tests, rows of tubes containing lactose broth are inoculated with water samples measuring 10 ml, 1 ml, and 0.1 ml. During incubation, coliform organisms, if present, would produce gas. A small inverted tube (Durham tube) is placed in each test tube to capture as gas is produced by coliform bacteria. Durham tubes float when filled with gas. The number of test tubes in each row, showing definite signs of gas production is identified and a statistical table is used to arrive at the point estimate and range of coliform bacteria in the sample.
The bacterial endotoxin test (BET), based on the fact that the blood of the horseshoe crab (Limulus or Tachypleus) gels or clots when it comes in contact with endotoxin. The Limulus amebocyte lysate (LAL) reagents are used to test for endotoxin using gel-clot method.
BioLab - Scope of Service
Sl | SvCd | Parameter | Test Material | Test Method |
---|---|---|---|---|
SPC | Standard Plate Count | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water. | IS 1622 Nutrient agar & 24h incubation at 37℃. | |
AMC | Aerobic Microbial Count | Water from Purifiers. | IS 14543 read with IS 5402: Plate count agar & 24h incubation at 37℃. | |
TVC | Total Viable Count | Water for Dialysis, Purified Water. | IS 23500 part 3 Tryptic soy agar & 48h incubation at 35-37℃. | |
HPC | Heterotrophic Plate Count | Drinking Water, Package Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water. | IS 5402 Plate count agar & 72h incubation at 30℃. | |
BET | Bacterial Endotoxin Assay (LAL) | Water for Dialysis, Purified Water. | Indian Pharmacopoea | |
MPNTC | MPN (Total Coliforms) | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water, Wastewater. | IS 1622 3.3.1 Multiple tube dilution test (MDT) | |
TTC | Thermotolerant Coliforms | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water, Wastewater. | IS 1622 - 3.3.3.2 subculture, from one presumptive positive tube of coliform test, in BGLB at 44℃ for 24h. | |
ECOLI | E. coli | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water, Wastewater. | IS 1622 - 3.3.4 subculture, from one presumptive positive tube of coliform test, in tryptone water at 44℃ for 24h. | |
MPNTTC | MPN (Thermotolerant Coliforms) | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water, Wastewater. | IS 1622 - 3.3.3.2 subculture, from all presumptive positive tubes of coliform test, in BGLB at 44℃ for 24h. | |
MPNEC | MPN (E. coli) | Drinking Water, Water for Food Processing, Water for Dialysis, Purified Water, Groundwater, Irrigation Water, Surface Water, Swimming Pool Water, Wastewater. | IS 1622 - 3.3.4 subculture, from all presumptive positive tubes of coliform test, in tryptone water at 44℃ for 24h. | |
Notes: |
Water Sample Collection Bottles for Bacteriological Analysis:
SBCd | Bottle Name | Details | Volume |
---|---|---|---|
CSB | Clean Sterile Bottle | Clean sterile rigid polypropylene bottle without any additive. | 250ml |
SBT | Sterile Bottle with Thiosulfate | Sterile rigid polypropylene bottle with sodium thiosulfate to neutralise chlorine, for bacteriological analysis (coliform test) of water. | 250ml |
SBTE | Sterile Bottle with Extra Thiosulfate | Sterile polypropylene bottle with extra sodium thiosulfate to neutralise excess chlorine, for bacteriological analysis of pool water. | 250ml |
NPT | Non Pyrogenic Tube | Non-pyrogenic glass tube to collect water sample for bacterial endotoxin test (BET) i.e. LAL assay. | 14ml |
Notes: |