Procedures
Selection of sample size: Size of sample will be governed by expected bacterial density and, if applicable, by regulatory requirements. In drinking water analyses, sample size will be limited only by the degree of turbidity or by the noncoliform growth on the medium.
An ideal sample volume will yield 20 to 80 total coliform colonies and not more than 200 clonied of all types (typical, atypical, and noncoliform background colonies) on a membrane filter surface. Analyze drinking waters by filtering 100 to 1000 mL, or by filtering replicate smaller sample volumes (e.g., duplicate 50-mL portions or four replicates of 25-mL portions). Analyze other water by filtering three different volumes (diluted or undiluted), depending on the expected bacterial density. (forpreparation of dilutions) When less than 10 mL, of sample is to be filtered, add approximately 10 mL. steril buffered dilution water to the funnel and then add sanple followed by another 25 to 50 mL dilution water before filtration or pipet the sample volume into sterile dilution water and then filter the contents of the dilutionbottle. This increase in water volume in uniform dispersion of the bacterial suspension over the affective filtering surface.