Potential Probiotics Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 Co-Aggregate with Clinical Isol

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Potential Probiotics Bacillus subtilis KATMIRA1933 and Bacillus amyloliquefaciens B-1895 Co-Aggregate with Clinical Isolates of Proteus mirabilis and Prevent Biofilm Formation Ammar Algburi 1 & Sarah A. Alazzawi 2 & Ali Ibrahim Ali Al-Ezzy 3 & Richard Weeks 4 & Vladimir Chistyakov 5 & Michael L. Chikindas 4,6

# Springer Science+Business Media, LLC, part of Springer Nature 2020

Abstract A urinary tract infection (UTI) is a multi-factorial disease including cystitis, pyelonephritis, and pyelitis. After Escherichia coli, Proteus mirabilis is the most common UTI-associated opportunistic pathogen. Antibiotic resistance of bacteria and infection recurrence can be connected to biofilm formation by P. mirabilis. In this study, human and sheep isolates of P. mirabilis were investigated for antibiotic sensitivity using an antibiotic disk test. Co-aggregation of the tested potential probiotic bacilli, Bacillus amyloliquefaciens B-1895 and Bacillus subtilis KATMIRA1933, with the isolated pathogen was also evaluated. Then, the antibiofilm activity of naturally derived metabolites, such as subtilin and subtilosin, in the bacilli-free supernatants was assessed against biofilms of P. mirabilis isolates. The isolated pathogens were sensitive to 30 μg of amikacin and 5 μg of ciprofloxacin but resistant to other tested antibiotics. After 24 h, auto-aggregation of B. amyloliquefaciens B-1895 was at 89.5% and higher than auto-aggregation of B. subtilis KATMIRA1933 (59.5%). B. amyloliquefaciens B-1895 strongly co-aggregated with P. mirabilis isolates from human UTIs. Cell-free supernatants of B. amyloliquefaciens B-1895 and B. subtilis KATMIRA1933 showed higher antimicrobial activity against biofilms of P. mirabilis isolated from humans as compared with biofilms of sheep isolates. According to our knowledge, this is the first report evaluating the anti-biofilm activity of probiotic spore-forming bacilli against clinical and animal UTI isolates of P. mirabilis. Further studies are recommended to investigate the anti-biofilm activity and the mode of action for the antimicrobial substances produced by these bacilli, subtilosin and subtilin. Keywords Probiotics . Bacillus . Co-aggregation . Proteus mirabilis . Biofilm prevention

Introduction

* Ammar Algburi [email protected]; [email protected] 1

Department of Biotechnology, Science College, University of Diyala, Baqubah, Iraq

2

Department of Microbiology, Veterinary College, University of Diyala, Baqubah, Iraq

3

Department of Pathology, College of Veterinary Medicine, University of Diyala, Baqubah, Iraq

4

Health Promoting Naturals Laboratory, School of Environmental and Biological Sciences, Rutgers State University, New Brunswick, NJ, USA

5

Southern Federal University, Rostov-on-Don, Russia

6

Don State Technical University, Rostov-on-Don, Russia

Urinary tract infection (UTI) is a multi-factorial disease including cystitis, pyelonephritis, and pyelitis [1, 2]. Several pathogens have been isolated from patients with a UTI, with Escherichia coli, Prote