Factors affecting enumeration and isolation of actinomycetes from Chesapeake Bay and Southeastern Atlantic Ocean sediments

Studies of the seasonal distribution of heterotrophic bacteria in Chesapeake Bay (USA) led us to the observation that fungizone was selective for actinomycetes without appreciably altering the total viable aerobic, heterotrophic bacterial counts. A study of the actinomycete populations in the Bay was undertaken, with several media compared for ability to enrich for actinomycetes: basal, casein, Czapek, starch-casein, glycerol-glycine, chitin and oil agar No. 2. Between 40 and 100% of the colonies appearing on chitin and oil No. 2 media were found to be actinomycetes, compared with <2% for the other media employed. It was also found that if sediment samples are frozen, a practice followed when laboratory facilities are not available in the field, a decrease in the total counts and in the actinomycete colony counts occurred. However, on a proportional basis, a slight increase in the percent actinomycetes was noted. A minimum of 21 days incubation time is recommended for recovery of actinomycetes. Also, the absence of NaCl from the actinomycete isolation media and incubation temperatures ≥15°C should give good recoveries of actinomycetes. A seasonal distribution of actinomycetes was noted for Chesapeake Bay, with maximal populations of actinomycetes observed in March and November. Actinomycetes were isolated in the ocean from depths ranging from 7 to 7,790 m. Greater numbers of actinomycetes were isolated using an incubation temperature of 25°C for the estuarine stations and 15°C for the deep-ocean stations. Greater numbers of actinomycetes were associated with polluted environments close to shore.