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• W207362019 abstract "Address(es): Elif Demirkan Uludag University, Faculty of Arts and Sciences, Biology Department, 16059, Gorukle, Nilufer-Bursa, Turkey, phone number: +90 224 29411794. *Corresponding author: edemirkan@uludag.edu.tr ABSTRACTARTICLE INFO Keywords: Bacillus, screening, antibiotic activity, sporulation INTRODUCTION Antibiotics have been used against infectious bacteria and fungi for over 50 years. Antibiotics are used as antitumor agents, immunosuppressive agents, hypocholesterolemic agents, enzyme inhibitors, antimigraine agents, and antiparasitic agents. Antibiotics first became widely available in the 1940s with the use of penicillin and sulfonamides. Since that time, the pharmaceutical industry has developed more than 100 varieties of these drugs, with 150 million prescriptions being written for antibiotics annually in the United States alone. This growth in antibiotic use has been paralleled by the ability of bacteria to resist being killed by these agents, which has resulted in a steady decline in the number of effective antibiotics each year (Levy, 1998). The emergence in recent years of numerous strains of pathogenic microorganisms that have developed resistance to a range of formerly efficacious antibiotics constitutes a serious threat to public health. The fact that certain nosocomial pathogens are already resistant to all available antibiotics and are therefore essentially untreatable dramatically demonstrates the need for completely new types of antibiotics (Conlon et al., 2006). Antibiotic activity is a feature of several kinds of soil bacteria and fungi and may represent a survival mechanism whereby organisms can eliminate competition and colonize a niche (Jensen and Wright, 1997; Talaro and Talaro, 1996). Bacillus species are one of the largest sources of bioactive natural products, exhibiting a wide range of antibiotic activities. There are 167 antibiotics produced by bacilli, including 66 derived from B. subtilis, 23 from B. brevis and the remainder from other species of the genus Bacillus. The main antibiotic producers of this genus are B. brevis (e.g., gramicidin and tyrothricin), B. cereus (e.g., cerexin and zwittermicin), B. circulans (e.g., circulin), B. laterosporus (e.g., laterosporin), B. licheniformis (e.g., bacitracin), B. polymyxa (e.g., polymyxin and colistin), B. pumilus (e.g., pumulin), and B. subtilis (e.g., polymyxin, difficidin, subtilin, mycobacillin, and bacitracin). Several research reports that in bacilli, the polypeptide antibiotics produced affect spore formation directly or indirectly (Vitkovic and Sadoff, 1975; Sarkar et al., 1977; Demain and Piret, 1978; Marahiel et al., 1979; Piret et al., 1983), and antibiotic activity by Bacillus begins at the early stages of the stationary phase, which coincides with the beginning of the sporulation process. Secondary metabolites are synthesized by a wide variety of pathways, and both the particular genetic make up of the producing strains and different environmental conditions can affect their production. Fermentation parameters, such as the incubation time, temperature, pH and available nutrients, can be modified to expand the range of the secondary metabolites produced (Furtado et al., 2005). Therefore, the purpose of this study was to isolate new, potentially antibiotic-activity bacteria from soil samples, optimize different parameters for antibiotic activity, and determine the relationship between antibiotic activity and sporulation. MATERIAL AND METHODS Isolation of potential antibiotic-activity bacteria Soil samples were collected from various cities of Turkey and were examined for the potential presence of antibiotic-activity bacteria. The test microorganisms, including Shigella sonnei (ATCC 9290), Salmonella typhimurium (ATCC 14028), Yersinia enterocolitica (ATCC 9610), Klebsiella pneumoniae (ATCC 13883), and Staphylococcus aureus (ATCC 25923), were inoculated onto nutrient agar plates. The soil samples were sprinkled on the lawn of test microorganisms. The plates were incubated at 37°C for 24 hours. After the incubation, the plates were analyzed for the inhibition zones surrounding the bacterial colonies. Identification of Bacillus species The isolated strains were identified by their morphological and biochemical characteristics according to Bergey’s Manual of Determinative Bacteriology (Buchanan and Gibbons, 1974). Cross-streak Method The preliminary screening was performed using the cross-streak method to test for antibiotic activity against Shigella sonnei (ATCC 9290), Salmonella typhimurium (ATCC 14028), Yersinia enterocolitica (ATCC 9610), Klebsiella pneumoniae (ATCC 13883) and Staphylococcus aureus (ATCC 25923). The isolated Bacillus strains were streaked across one third of the plate, and the plate was incubated at 37°C for 24 h. After the Bacillus grew, the test bacteria were streaked perpendicular to the Bacillus, and the plate was incubated at 37°C for another 24 h. The microbial inhibition was evaluated by determining the diameter of the inhibition zones. Fifty-two Bacillus strains, which were isolated from different soil samples, were screened for antibiotic properties. The Bacillus strains were checked for antibacterial properties by the cross-streak method against 5 test pathogens, and 25 Bacillus strains had an effect on the test microorganisms. One strain of Bacillus, which exhibited the largest inhibition zone (25 mm) against Shigella sonnei, was named Bacillus sp. EA62. The antibacterial activity from Bacillus sp. EA62 was tested in six different culture media against Shigella sonnei using the agar well diffusion method. The best activity medium was selected and used for further studies. The influence of the incubation period, pH, and different glucose and nitrogen concentrations on the antibacterial activity was studied. The optimal conditions for the strongest antibiotic activity were found to be 72 hours (18 mm), pH 7.5 (23 mm), 3% glucose (25 mm), and 0.3% nitrogen concentration (23 mm). Additionally, the relationship between the antibiotic activity and sporulation was investigated. Accordingly, it was determined that the increase of the activity paralleled sporulation. Received 10. 1. 2013 Revised 21. 2. 2013 Accepted 21. 2. 2013 Published 1. 4. 2013 Regular article" @default.
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• W207362019 title "The effect of growth parameters on the antibiotic activity and sporulation in Bacillus spp. isolated from soil." @default.
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