The predatory potential of Bdellovibrio bacteriovorus against clinically isolated pathogens with extensively drug-resistance (XDR)

Subject Areas : Applied Microbiology

Salman Odooli ¹ , Rasoul Roghanian ² , Giti Emtiazi ³ , Milad Mohkam ⁴ , Younes Ghasemi ⁵

1 - Ph.D Student in Microbiology, Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Sciences and Technology, University of Isfahan, Isfahan, Iran.
2 - Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Sciences and Technology, University of Isfahan, Isfahan, Iran,.
3 - Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Sciences and Technology, University of Isfahan, Isfahan, Iran.
4 - Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
5 - Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.

Received: 2020-01-24 Accepted : 2020-02-05 Published : 2020-02-20

Keywords: identification, antimicrobial resistance, Bacteriolytic activity, Bdellovibrio, Predation,

Abstract :

Background & Objectives: Bdellovibrio-and-like organisms (BALOs) are a group of predatory bacteria which invade other Gram-negative bacterial cells for growth. The bacteriolytic nature of Bdellovibrios make them one of the promising alternatives for conventional antibiotics. In this study, the isolation and molecular identification of Bdellovibrio bacteriovorus strain SOIR-1 was described. The antibiotic resistance pattern of some clinically isolated Gram-negative pathogens was determined, and the predatory potency of SOIR-1 toward them was evaluated. Material & Methods: Double-layer agar technique, transmission electron microscopy, and PCR targeting the Bdellovibrios-specific hit locus were used for the isolation, morphological investigation, and molecular identification of SOIR-1, respectively. Following the antibiotic resistance profile determination of clinical isolates, the bacteriolytic activity of SOIR-1 against them was evaluated through the plaque formation assay and lysis analysis in the broth co-cultures. Results: SOIR-1 was identified as a strain of Bdellovibrios bacteriovorus through the transmission electron microscopy examination and specific PCR detection. All clinical isolates showed the properties of extensively drug-resistant (XDR) and typical Bdellovibrios plaques were developed on their lawns of cells. The SOIR-1 had the highest and lowest predation efficiency among the clinical isolates toward Acinetobacter baumannii (84.33%) and Pseudomonas aeruginosa-369 (55.16%), respectively. Conclusion: This study highlights the great potential of SOIR-1 to prey and lyse XDR pathogens, regardless of their antimicrobial resistance state. So, B. bacteriovorus can be considered as a living antibiotic in the cases of infections caused by multidrug-resistant bacteria.

References:

1. Davies J, Davies D. Origins and evolution of antibiotic resistance. Microbiol Mol Biol Rev.
   2010;74(3):417-33.
   2. Laws M, Shaaban A, Rahman KM. Antibiotic resistance breakers: current approaches and
   future directions. FEMS Microbiol Rev. 2019;43(5):490-516.
   3. Aslam B, Wang W, Arshad MI, Khurshid M, Muzammil S, Rasool MH, et al. Antibiotic
   resistance: a rundown of a global crisis. Infect Drug Resist. 2018;11:1645.
   4. Nikaido H. Multidrug resistance in bacteria. Annu Rev Biochem. 2009;78:119-46.
   5. Abat C, Fournier P-E, Jimeno M-T, Rolain J-M, Raoult D. Extremely and pandrug-resistant
   bacteria extra-deaths: myth or reality? Eur J Clin Microbiol Infect Dis. 2018;37(9):1687-97.
   6. Magiorakos AP, Srinivasan A, Carey R, Carmeli Y, Falagas M, Giske C, et al.
   Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international
   expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect.
   2012;18(3):268-81.
   7. Bassetti M, Peghin M, Vena A, Giacobbe DR. Treatment of infections due to MDR
   Gram-negative bacteria. Front Med. 2019;6.
   8. Vivas R, Barbosa AAT, Dolabela SS, Jain S. Multidrug-resistant bacteria and alternative
   methods to control them: An overview. Microb Drug Resist. 2019;25(6):890-908.
   9. Rios AC, Moutinho CG, Pinto FC, Del Fiol FS, Jozala A, Chaud MV, et al. Alternatives to
   overcoming bacterial resistances: state-of-the-art. Microbiol Res. 2016;191:51-80.
   10. Parisien A, Allain B, Zhang J, Mandeville R, Lan C. Novel alternatives to antibiotics:
   bacteriophages, bacterial cell wall hydrolases, and antimicrobial peptides. J Appl Microbiol.
   2008;104(1):1-13.
   11. Wu H, Lu H, Huang J, Li G, Huang Q. EnzyBase: a novel database for enzybiotic studies.
   BMC Microbiol. 2012;12(1):54.
   12. Kadouri DE, To K, Shanks RM, Doi Y. Predatory bacteria: a potential ally against
   multidrug-resistant Gram-negative pathogens. PloS one. 2013;8(5):e63397.
   13. Martin MO. Predatory prokaryotes: an emerging research opportunity. J Mol Microbiol
   Biotechnol. 2002;4(5):467-78.
   14. Markelova NY. Predacious bacteria, Bdellovibrio with potential for biocontrol. Int J Hyg
   Environ Health. 2010;213(6):428-31.
   15. Dwidar M, Monnappa AK, Mitchell RJ. The dual probiotic and antibiotic nature of
   Bdellovibrio bacteriovorus. BMB Rep. 2012;45(2):71-8.
   
   16. Davidov Y, Jurkevitch E. Diversity and evolution of Bdellovibrio-and-like organisms
   (BALOs), reclassification of Bacteriovorax starrii as Peredibacter starrii gen. nov., comb. nov.,
   and description of the Bacteriovorax–Peredibacter clade as Bacteriovoracaceae fam. nov. Int J
   Syst Evol Microbiol. 2004;54(5):1439-52.
   17. Strauch E, Schwudke D, Linscheid M. Predatory mechanisms of Bdellovibrio and like
   organisms. Future Microbiol. 2007;2(2):63-73.
   18. Sockett RE, Lambert C. Bdellovibrio as therapeutic agents: a predatory renaissance? Nat Rev
   Microbiol. 2004;2(8):669.
   19. Stolp H, Starr M. Bdellovibrio bacteriovorus gen. et sp. n., a predatory, ectoparasitic, and
   bacteriolytic microorganism. Antonie Van Leeuwenhoek. 1963;29(1):217-48.
   20. Pan A, Chanda I, Chakrabarti J. Analysis of the genome and proteome composition of
   Bdellovibrio bacteriovorus: indication for recent prey-derived horizontal gene transfer.
   Genomics. 2011;98(3):213-22.
   21. Rendulic S, Jagtap P, Rosinus A, Eppinger M, Baar C, Lanz C, et al. A predator unmasked:
   life cycle of Bdellovibrio bacteriovorus from a genomic perspective. Science. 2004;303
   (5658):689-92.
   22. Lambert C, Morehouse KA, Chang C-Y, Sockett RE. Bdellovibrio: growth and development
   during the predatory cycle. Curr Opin Microbiol. 2006;9(6):639-44.
   23. Sockett RE. Predatory lifestyle of Bdellovibrio bacteriovorus. Annu Rev Microbiol.
   2009;63:523-39.
   24. Van Essche M, Quirynen M, Sliepen I, Loozen G, Boon N, Van Eldere J, et al. Killing of
   anaerobic pathogens by predatory bacteria. Mol Oral Microbiol. 2011;26(1):52-61.
   25. Shanks RM, Kadouri DE. Predatory prokaryotes wage war against eye infections. Future
   Microbiol. 2014;9(4):429-32.
   26. Monnappa AK, Dwidar M, Seo JK, Hur J-H, Mitchell RJ. Bdellovibrio bacteriovorus inhibits
   Staphylococcus aureus biofilm formation and invasion into human epithelial cells. Sci Rep.
   2014;4:3811.
   27. Kadouri D, O'Toole GA. Susceptibility of biofilms to Bdellovibrio bacteriovorus attack. Appl
   Environ Microbiol. 2005;71(7):4044-51.
   28. Dashiff A, Junka R, Libera M, Kadouri D. Predation of human pathogens by the predatory
   bacteria Micavibrio aeruginosavorus and Bdellovibrio bacteriovorus. J Appl Microbiol.
   2011;110(2):431-44.
   29. Chu WH, Zhu W. Isolation of Bdellovibrio as biological therapeutic agents used for the
   treatment of Aeromonas hydrophila infection in fish. Zoonoses Public Health. 2010;57(4):
   258-64.
   30. Dharani S, Kim DH, Shanks RM, Doi Y, Kadouri DE. Susceptibility of colistin-resistant
   pathogens to predatory bacteria. Research in microbiology. 2018;169(1):52-5.
   31. Markelova NO. The Potential of Bdellovibrio For the Biocontrol of the Infectious Agent
   Vibrio cholerae. 2015.
   32. Dashiff A, Kadouri D. Predation of oral pathogens by Bdellovibrio bacteriovorus 109J. Mol
   Oral Microbiol. 2011;26(1):19-34.
   33. Rajababu P, Vikram RG, Jagadish RG, Rathi E. Periodontal predators preying the pathogens.
   Indian J Dent Res. 2013;5(2):1182-6.
   34. Loozen G, Boon N, Pauwels M, Slomka V, Herrero ER, Quirynen M, et al. Effect of
   Bdellovibrio bacteriovorus HD100 on multispecies oral communities. Anaerobe. 2015;35:
   45-53.
   35. Cao H, He S, Wang H, Hou S, Lu L, Yang X. Bdellovibrios, potential biocontrol bacteria
   against pathogenic Aeromonas hydrophila. Vet Microbiol. 2012;154(3-4):413-8.
   36. Shanks RM, Davra VR, Romanowski EG, Brothers KM, Stella NA, Godboley D, et al. An eye
   to a kill: using predatory bacteria to control Gram-negative pathogens associated with ocular
   infections. PloS one. 2013;8(6):e66723.
   37. Hope D, Ampaire L, Oyet C, Muwanguzi E, Twizerimana H, Apecu RO. Antimicrobial
   resistance in pathogenic aerobic bacteria causing surgical site infections in Mbarara regional
   referral hospital, Southwestern Uganda. Sci Rep. 2019;9(1):1-10.
   38. CLSI. Performance standards for antimicrobial susceptibility testing. 30th ed. CLSI
   supplement M100. Wayne, PA: Clinical and laboratory standards institute; 2020.
   39. Jurkevitch E. Isolation and classification of Bdellovibrio and like organisms. Curr Protoc
   Microbiol. 2012;26(1):7B. 1.-7B. 1.20.
   40. Iebba V, Santangelo F, Totino V, Nicoletti M, Gagliardi A, De Biase RV, et al. Higher
   prevalence and abundance of Bdellovibrio bacteriovorus in the human gut of healthy subjects.
   PloS one. 2013;8(4):e61608.
   41. Schwudke D, Strauch E, Krueger M, Appel B. Taxonomic studies of predatory bdellovibrios
   based on 16S rRNA analysis, ribotyping and the hit locus and characterization of isolates from
   the gut of animals. Syst Appl Microbiol. 2001;24(3):385-94.
   42. Rogosky AM, Moak PL, Emmert EA. Differential predation by Bdellovibrio bacteriovorus
   109J. Curr Microbiol. 2006;52(2):81-5.
   43. Li H, Liu C, Chen L, Zhang X, Cai J. Biological characterization of two marine
   Bdellovibrio-and-like organisms isolated from Daya bay of Shenzhen, China and their
   application in the elimination of Vibrio parahaemolyticus in oyster. Int J Food Microbiol.
   2011;151(1):36-43.
   44. Varon M, Shilo M. Attachment of Bdellovibrio bacteriovorus to cell wall mutants of
   Salmonella spp. and Escherichia coli
   Journal of bacteriology. 1969;97(2):977.
   45. Strauch E, Beck S, Appel B. Bdellovibrio and like organisms: potential sources for new
   biochemicals and therapeutic agents? Predatory Prokaryotes: Springer; 2006. p. 131-52.
   46. Koval S, Hynes S. Effect of paracrystalline protein surface layers on predation by Bdellovibrio
   bacteriovorus. Journal of bacteriology. 1991;173(7):2244-9.
   47. Koval SF, Bayer ME. Bacterial capsules: no barrier against Bdellovibrio. Microbiology.
   1997;143(3):749-53.
   48. Lambert C, Hobley L, Chang C-Y, Fenton A, Capeness M, Sockett L. A predatory patchwork:
   membrane and surface structures of Bdellovibrio bacteriovorus. Adv Microb Physiol.
   2008;54:313-61.
   49. Duncan MC, Forbes JC, Nguyen Y, Shull LM, Gillette RK, Lazinski DW, et al. Vibrio
   cholerae motility exerts drag force to impede attack by the bacterial predator Bdellovibrio
   bacteriovorus. Nat Commun. 2018;9(1):4757.
   50. Sun Y, Ye J, Hou Y, Chen H, Cao J, Zhou T. Predation efficacy of Bdellovibrio bacteriovorus
   on multidrug-resistant clinical pathogens and their corresponding biofilms. Jpn J Infect Dis.
   2017;70(5):485-9.
   51. Gupta S, Tang C, Tran M, Kadouri DE. Effect of predatory bacteria on human cell lines. PloS
   one. 2016;11(8):e0161242.
   52. Schwudke D, Linscheid M, Strauch E, Appel B, Zahringer U, Moll H, et al. The obligate
   predatory Bdellovibrio bacteriovorus possesses a neutral lipid A containing alpha-D-Mannoses
   that replace phosphate residues: similarities and differences between the lipid As and the
   lipopolysaccharides of the wild type strain B. bacteriovorus HD100 and its host-independent
   derivative HI100. J Biol Chem. 2003;278(30):27502.
   53. Atterbury RJ, Hobley L, Till R, Lambert C, Capeness MJ, Lerner TR, et al. Effects of orally
   administered Bdellovibrio bacteriovorus on the well-being and Salmonella colonization of
   young chicks. Appl Environ Microbiol. 2011;77(16):5794-803.
   54. Shatzkes K, Chae R, Tang C, Ramirez GC, Mukherjee S, Tsenova L, et al. Examining the
   safety of respiratory and intravenous inoculation of Bdellovibrio bacteriovorus and Micavibrio
   aeruginosavorus in a mouse model. Sci Rep. 2015;5:12899.
   55. Shatzkes K, Singleton E, Tang C, Zuena M, Shukla S, Gupta S, et al. Predatory bacteria
   attenuate Klebsiella pneumoniae burden in rat lungs. MBio. 2016;7(6):e01847-16.
   56. Shemesh Y, Jurkevitch E. Plastic phenotypic resistance to predation by Bdellovibrio and like
   organisms in bacterial prey. Environ Microbiol. 2004;6(1):12-8