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The search for new antibiotics continues in a rather overlooked hunting ground. In the course of screening for new antibiotic-producing microorganisms, isolates showing antimicrobial activity were isolated from waste soil samples from various habitats in the Industrial Areas in Dheradun, Uttarakhand, India. Existing methods of screening for antibiotic producers together with some novel procedures were reviewed. Both modified agar-streak and agar-plug methods were used in the primary screens. The use of selective isolation media, with or without antibiotic incorporation and/or heat pretreatment, enhanced the development of certain actinomycete colonies on the isolation plates. Antibiotics have long been considered the "magic bullet" that would end infectious disease. Although they have improved the health of countless numbers of humans and animals, many antibiotics have also been losing their effectiveness since the beginning of the antibiotic era. Bacteria have adapted defenses against these antibiotics and continue to develop new resistances, even as we develop new antibiotics. In recent years, much attention has been given to the increase in antibiotic resistance. As more microbial species and strains become resistant, many diseases have become difficult to treat, a phenomenon frequently ascribed to both indiscriminate and inappropriate use of antibiotics in human medicine. However, the use of antibiotics and antimicrobials in raising food animals has also contributed significantly to the pool of antibiotic resistant organisms globally and antibiotic resistant bacteria are now found in large numbers in virtually every ecosystem on earth. Dual culture bioassays were used to screen seven selected Bacillus isolates for activity against four plant pathogenic fungi in vitro. All isolates were able to inhibit the pathogens to varying degrees. Two isolates, R29 and B81, were selected for further testing and characterization. Further bioassays were performed on five complex nutrient media which were adjusted to pH S.S and 7, and both incubated at 2SoC and 30°C" respectively. It was found that pH and media composition showed significant influences on the antifungal activities of the isolates tested, but that a SoC temperature difference in incubation temperature did not. Tryptone soy agar was found to give rise to the largest inhibition zones. Both isolates were tentatively identified using standard biochemical and morphological tests. Based on its phenotypic characteristics, R29 was identified as a strain of B. subtilis. B81 proved to be more difficult to assign to a specific group or species of Bacillus, though B. subtilis and B. licheniformis were considered to be the nearest candidates. Genomic DNA was extracted from both isolates and a portion of each of their 16s rDNA genes were amplified and sequenced for homology testing against the GeneBank database. Homology testing confirmed that both isolates were members of the genus Bacillus and most probably strains of B. subtilis. The DNA fragment used for sequencing proved to be too small to give conclusive identification of the isolates.

“Isolation and Biochemical Characterization of Antibiotic Producing Microorganism from Waste Soil Samples of Certain Industrial areas of India”