VII.

BACTERIOLOGIC ANALYSIS OF WATER

1. What are the 3 most important stages in water analysis?

- Presumptive test

o Inoculate undiluted water sample into DSLB and SSLB

o 0.1ml water sample + two SSLB tubes
o 1ml water sample + two SSLB tubes.
o 10ml water sample + 5 DSLB tubes
o Mix tubes by rolling the tube between the palm of the hands
o Incubate overnight at 37C
o Presence of gas formation indicates lactose fermentation and is presumptive for coliforms.

- Confirmatory test

o Select lactose broth showing the largest amount of gas formation using aseptic technique, fish 1 loopful of
sample and inoculate the EMB or Endo agar.
o Use clock-streak
o Incubate at 37C for 18-24hrs
o Observe characteristics of the growth
o Presence of E.coli (greenish metallic sheen) is confirmatory of fecal contamination.

- Completed test

o Select an isolated colony from the EMB

o Using a septic technique, fish a colony and inoculate on NA slant and another SSLB
o Incubate at 37C for 18-24hrs
o Observe gas formxn inverted tube
o Make smear from agar slant, gram stain and examine
o Coliforms are gram neg

2. What is the normal colony count in water?

– Safe for drinking is <500 CFU/ml

– Purified water at 100 CFU/ml

3. What is the significance of high colony count in water?

- Presence of coliforms meaning the water is not safe fro drinking.

- Temperate zone: Salmonella typhi and Salmonella Typhimurium
- Tropical areas: Vibrio cholera

4. How do you identify the pollutant in water?

- Culture: EMB, MC
- Biochemical test: IMVIC
- Gram staining