Prototheca zopfii was isolated from cow-composite milk and bulk tank milk by a new selective Prototheca enrichment method. Repeated testing of cow-composite milk from individual cows resulted in P. zopfii isolation data indicating a strong statistical correlation of P. zopfii with specific cows. Prototheca sp. were isolated from the milk of 31 of 79 cows in a single herd, and contamination was discounted as the source. Prototheca sp. were also recovered from 28 of 787 bulk tank milk samples from the Eastern U.S.A., and 22 of 69 temperature control milk samples from pooled dairy farm milk from delivery trucks.