Generally, hot areas such as hot springs have high potential for growth of thermophilic bacteria. These bacteria are very useful in various industries in the fields of microbiology and biotechnology. One of the most important and most diverse groups of thermophiles is Bacillaceae family. Many species of this family are Extremophiles. Recently, scientists have found a tendency to study about thermophilic bacteria, thus they are good sources of biotechnology for industrial products of heat-stable enzymes. In this study, the water samples from hot spring Sehezar in Iran, were enriched and cultivated in the modified medium mBTM under sub-merged special conditions. Then, phenotyping and genotyping characteristics of single colonies have been determined by morphological, biochemical and molecular tests. According to the results obtained, isolated colonies were from two thermophilic genera of Bacillus and Geobacillus. Several tests were used to identify bacteria such as API, PCR-RFLP, 16s rRNA and HRM. In conclusion, this study led to the isolation and identification of one new species and a few new strains. In addition, species of thermophilic bacteria were isolated and identified by cultivation methods in a selective media, to finally be used in molecular and biochemical methods for detection and classification.

Keywords: Thermophiles; Extremophiles; mBTM; API; HRM