ASSESSMENT OF PROFILE DEPTH, SITE OF SAMPLING, TYPE OF MEDIA AND METHODS USED FOR THE ISOLATION OF ACTINOMYCETES

ALGAFARI R.N.¹*
¹Biotechnology Research Center, Al-Nahrain University, Baghdad, Iraq.
* Corresponding Author : rebahalgafari@gmail.com

Received : 16-05-2014     Accepted : 10-07-2014     Published : 31-07-2014
Volume : 6     Issue : 1       Pages : 553 - 558
Int J Microbiol Res 6.1 (2014):553-558

In this work, we investigated the critical conditions affecting isolation of actinomycetes family members. Sites thought to be rich with these bacteria were river banks, farming soils, animal farms, animal manure, bird dung, slaughter houses, and diesel contaminated soils. These were site on which sampling, profile depth, isolation procedure, and cultivation medium were tested. All these sites gave a considerable number of isolates with different criteria. The profile depth of sampling was a critical factor for the isolation. The depth of 10 cm and surface scratches were the best for such purpose. Method of isolation was found to affect isolation procedure dramatically. For the isolation of bio-active actinomycetes, the use of procedures with selective agent like phenol is recommended, whereas, using CaCo3 for this purpose in some procedures was effective in isolating distinct and separated colonies of actinomycetes. Media such as ISP4 and ISP2 are highly recommended for cultivation of newly isolated colonies, since such media were able to enhance specific criteria such pigment formation and helped these colonies to reach maturation at considerable time of incubation. The use of antifungal agents during isolation procedure was very important to obtain uncontaminated colonies, but instead of adding these agents with culture media, we found adding these agents directly on the plates before adding diluted sample from soil was very effective in eliminating fungal growth.

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