International Research Journal of Pharmacy and Medical Sciences ISSN (Online): 2581-3277 15 Marwan Yousif Hussain, Adnan Ali-Nizam, Samir Abou-Isba, Ahed Abou-Younes, and Wajid Khaddour, ―A Broad-Spectrum Antibacterial Activity of Lyophilized Crude Extracts of Bacillus Subtilis against Clinical and Food-Borne Pathogens,‖ International Research Journal of Pharmacy and Medical Sciences (IRJPMS), Volume 1, Issue 3, pp. 15-21, 2018. A Broad-Spectrum Antibacterial Activity of Lyophilized Crude Extracts of Bacillus Subtilis against Clinical and Food-Borne Pathogens Marwan Yousif Hussain* 1 , Adnan Ali-Nizam 1 , Samir Abou-Isba 1 , Ahed Abou-Younes 2 , Wajid Khaddour 1 1 Department of Plant Biology- Faculty of Science- Damascus University- Syria 2 Department of Food Science- Faculty of Agriculture- Damascus University- Syria Email address: mrgoldeny@gmail.com Abstract— The aim of this research is to study the broad spectrum antibacterial activity of crude lyophilized extracts from Bacillus subtilis, then characterize these antibacterial activity. B. subtilis strains were isolated and identified by both API CHB 50 and molecular methods. Staphylococci strains, negative gram bacteria and Candida spp. were isolated from clinical and food samples and identified by biochemical and morphological tests. The crude antibacterial were extracted by flasks containing Lactose Broth LB, which incubated for 72 hours in shaker incubator at 37 º ºC at 150 rev/min, then centrifuged 12000 rev/min or 15 min. at 4 ºC and filtrated the supernatants through 0.45 μm then frozen and lyophilized. The crude extracts were run on Diamonsil C18 column (5 μm, 250 × 4.6 mm) in HPLC system to separate the antibiotics and run on FTIR spectrum. Some strains of B. subtilis showed high activity against target strains, and others did not. The antibacterial activity of crude lyophilized extracts from B. subtilis (BS1A, BS2D, BS7SH, BS8SH ) against S. aureus strains was between high (16-40 mm) and intermediate (8-14 mm). While the antibacterial activity was intermediate (8-14 mm) against Gram negative and Candida spp. strains. Two kinds of antibiotics were separated from lyophilized extracts of B. subtilis and identified to the peptides families, Bacitracin A and Polymyxin B. Keywords— Antibacterial, Staphylococci, Listeria, Candida, crude, peptide. I. INTRODUCTION acillus subtilis produces many antibacterial and antifungal peptides such as bacitracin and polymyxin and lipopeptide such as surfactin, iturin and fengycin with a great potential for biotechnological and biopharmaceutical applications (Schallmey et al., 2004; Stein, 2005; Sumi et al., 2015). Staphylococcus aureus especially methicillin resistance S. aureus (MRSA) and Coagulase Negative Staphylococci (CNS) are still one of the five most common causes of nosocomial infections, in addition, the infections are associated with surgical sites, wound infections, catheters and prosthetic implants (Chambers et al., 2012; Kainthola et al., 2015). Also Worldwide foodborne pathogens cause a lot of sufferings and deaths, mainly S. aureus and Listeria monocytogenes (Soni, 2015). Candida spp. particularly, Candida albicans remains the most common cause of invasive fungal infections, such as superficial, subcutaneous and systemic infections. Candidiasis has become a major problem in hospitals where cause death in hospitalized adults and children in many countries worldwide (Guinea, 2014; Hirota, 2016). Most research, are focused on the ability of Bacillus species to produce antibiotics and antifungals. Kuijk et al. (2011) studied and elucidated the molecular mechanism of action of the antimicrobial peptide subtilosin, which were extracted from Bacillus amyloliquefaciens against the foodborne pathogen L. monocytogenes Scott A, also Sethi et al. (2013) showed that B. subtilis which was isolated from the soil showed activity against S. aureus, Pseudomonas aeruginosa, and Escherichia coli, while Ndahi et al.(2013) studied the prevalence and antimicrobial sensitivity of L. monocytogenes and S. aureus which were isolated from raw meat and meat products, whereas in study of Ceresa et al.(2015) was appeared the ability of biosurfactants which were produced by Lactobacillus brevis was investigated to inhibit the biofilm formation of C. albicans, in addition in study of Haubert et al. (2015) about 50 isolates of L. monocytogenes were isolated from food and food environment and studied for antimicrobial resistance and genetic relationships, while in study of Das et al. (2016) it was evaluated the antimicrobial and antibiofilm of small molecules against S. aureus and Pseudomonas aeruginosa, also in study of Sharma and Sharma, (2017) Bacillus licheniformis which was isolated from indigenous traditional fermented beverage showed broad antagonism against most pathogens. However, the prevalence of antimicrobial resistance among key microbial pathogens is increasing at an alarming rate worldwide. This study presented the characterization of some new strains of B. subtilis which produce antibiotics and antifungal then were evaluated their activity against positive and negative bacteria and C. albicans. II. MATERIAL AND METHODS 2.1 Isolation and Identification of B. subtilis During the period between 6/2014 - 6/2015, three different soil samples were taken from each site from cultivated and B