Using autochthonous Bdellovibrio as a predatory bacterium for reduction of Gram-negative pathogenic bacteria in urban wastewater and reuse it

  • Neda Jafarian Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
  • Abbas Akhavan Sepahi Mail Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
  • Nafiseh Sadat Naghavi Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran
  • Farzaneh Hosseini Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
  • Jamileh Nowroozi Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran
Autochthonous Bdellovibrio;, Predatory bacterium;, Bio-control;, Gram-negative pathogenic bacteria;, Waste water treatment


Background and Objectives: The microbial contamination of wastewater is associated with health risks. The aim of this study was to use the autochthonous Bdellovibrio potential to prey Gram-negative pathogenic bacteria as a bio-control agent to treat urban wastewater.
Materials and Methods: Thirty-six raw sewage samples were collected for isolation of Bdellovibrio. Double layer plaque assay was used for isolation and the isolates were identified by microscopic examination and molecular analysis. To evaluate the predatory potential for decrease number of Gram-negative pathogenic bacteria, plaque perdition assay, reduction in host cells viability by colony-forming unit (CFU) counting, reduction in optical density (OD) in co-cultures and assay of killing efficiency were carried out. Also, the raw wastewater was treated by Bdellovibrio then the reduction in CFU counting and reduction in OD was evaluated.
Results: Four strains of Bdellovibrio were isolated and were registered in Gene Bank. Clear plaques were observed after 3-6 days of incubation for all prey cells. The CFU enumerations of all preys were decreased after 48 hrs in co-cultures and raw wastewater. Also, OD was decreased down to 0.2 nm after 48 hrs.
Conclusion: These autochthonous Bdellovibrio strains are proposed to use for bio-control of Gram-negative pathogenic bacteria in wastewater and reuse it for irrigation in arid regions.


1. Dwidar M, Mannappa A, Mitchell R. The dual probiotic and antibiotic nature of Bdellovibrio bacteriovorus. BMB Rep 2012; 45: 71-78.
2. Sinha A, Hurakadli M, Ravindra S, Agarwal A. Bdellovibrio and like organisms: The predatory ssassin. IOSR-JDMS 2014; 13: 32-36.
3. Markelova N. Interaction of Bdellovibrio bacteriovorus with bacteria Campylobacter jejuni and Helicobacter pylory. Microbiology 2010; 79: 777-779.
4. Ezzedine JA, Jacas L, Desdevises Y, Jacquet S. Bdellovibrio and like organisms: an unseen elephant in the room? Front Microbiol 2020; 11: 98.
5. Shanks R, Davar V, Romanowski E, Brothers K, Stella N, 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.
6. Dwidar M, Im H, Seo JK, Mitchel RJ. Attack-phase Bdellovibrio bacteriovorus responses to extracellular nutrients are analogous to those seen during late interperiplasmic growth. Microb Ecol 2017; 74:937-946.
7. Lu F, Cai J. The protective effect of Bdellovibrio and like organisms on tilapia fish fillets against Salmonella entrica spp. entrica serovar Typhimurium. Lett Appl Microbiol 2010; 51:625-631.
8. El-Shanshoury A, Abo-Amer A, Alzahrani O. Isolation of Bdellovibrio sp. from wastewater and their potential application in control of Salmonella paratyphi in water. Geomicrobiol 2016; 33: 886-893.
9. Ozkan M, Yilmaz H, Celik MA, Sengezer C, Erhan E, Keskinler B. Application of Bdellovibrio bacteriovorous for reducing fouling of membranes used for wastewater treatment. Turkish J Biochem 2018; 43: 296-305.
10. Calhau V, Mendes C, Pena A, Mondonca N, Dasilva G. Virulence and plasmidic resistance determinants of Echerichia coli from municipal and hospital wastewater treatment plants. J Water Health 2015; 13: 311.
11. Cheng HA, Lucy FE, Broaders MA, Mastitsky SE, Chen C, Murray A. Murray amunicipal wastewater treatment plants as pathogen removal system and as a contamination source of Noroviruses and Enterococcus faecalis. J Water Health 2012; 10: 380-389.
12. Oyedara OO, de Jesus De Luna-Santillana E, Olguin-Rodriguez O, Guo X, Mendoza-Villa MA, Menchaca-Arredondo JL, et al. Isolation of Bdellovibrio sp. from soil samples in Mexico and their potential applications in control of pathogens. MicrobiologyOpen 2016; 5: 992-1002.
13. Feng S, Tan, C, Cohen Y, Rice S. Isolation of Bdellovibrio bacteriovorus from tropical wastewater treatment plant and predation of mixed species biofilms assembled by the native community members. Environ Microbiol 2016; 18: 3923-3931.
14. Richard G, Fay J, Dickens K, Parent M, Soroka D, Boyd F. Predatory bacteria as natural modulators of Vibrio parahaemolyticus and Vibrio vulnificus in seawater and oysters. Appl Environ Microbiol 2012; 78: 7455-7466.
15. Passariello C, Schippa S, Nardo DD, Polimeni A, Testarelli L. Molecular detection of the predatory bacterium Bdellovibrio bacteriovorus from dental biofilms. Dent Hypotheses 2019;10:103.
16. Dashiff A, Junka R, Libera M, Kadouri D. Predation of human pathogens by the predatory bacteria Micavibrio aeroginosavorus and Bdellovibrio bacteriovorus. J Appl Microbiol 2011; 110: 431-444.
17. Sar T, Umeh E, Akosu D. Occurrence, detection and isolation of Bdellovibrio spps. from some fresh water bodies in Benue State, Nigeria. Microbiol J 2015; 5: 21-27.
18. Kongrueng J, Mitraparp P, Bangpanwimon K, Robins W, Vuddhakul V, Mekalanos J. Isolation of Bdellovibrio and like organisms and potential to reduce acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus. Dis Aquat Organ 2017; 124: 223-232.
19. Atterbury R, Hobley L, Till R, Capeness M, Lerner T, Fenton A, et al. Effects of orally administered Bdellovibrio bacteriovorus on the well-being and Salmonella colonization of young chicks. Appl Environ Microbiol 2011; 77: 5794-5803.
20. Russo R, Chae R, Mukherjee S, Singleton E, Occi J, Kadouri D, et al. Susceptibility of select A gents to predation by predatory bacteria. Microorganisms 2015; 3:903-912.
21. Bratanis E, Andersson T, Lood R, Bukowska-Fanlband B. Biotechnological potential of Bdellovibrio and like organisms and their secreted enzymes. Front Microbiol 2020; 11: 662.
22. Boileau M, Clinkenbeard K, Iandolo J. Assessment of Bdellovibrio bacteriovorus 109J killing of Moraxella bovis in an in vitro model of infectious bovine Keratoconjuctivitis. Can J Vet Res 2011; 75: 285-291.
23. Larrnic S, Mancini ML. Can constructed wetlands treat wastewater for reuse in agriculture? Review of guidelines and examples in South Erope. Water Sci Technol 2016; 73:2616-2626.
24. Ali S, Mahmood R, Muneer A, Khalil M, Sheikh N, Muhammad Tahir H, et al. Assessment of spring water microbiology and role of Typha angustata as biosorbent. Water Environ Res 2019; 91: 1705-1717.
25. Baresel C, Dalgern L, Almemark M, Lazic A. Environmental performance of wastewater reuses systems: impact of system boundaries and external conditions. Water Sci Technol 2016;73: 1387-1394.
26. Ren X, Zhang Y, Chen H. Greywater treatment technology and reuse of reclaimed water for toilet flushing. Environ Sci Pollut Res Int 2020; 27: 34653-34663.
27. Zafar Z, Akhtar J, Amjad M, Anwar-Ul-Haq M, Saqib ZA. Trace metals accumulation and antioxidants profiling in two maize genotypes against sewage and textile wastewater treatment. Clean-Soil Air Water 2019; 47: 1800063.
28. Barbagallos S, Luigi Cirelli G, Consoli S, Liciardello F, Marzo A, Toscano A. Analysis of treated wastewater reuses potential for irrigation in Sicily. Water Sci Technol 2012; 65: 2024-2033.
29. Periasamy D, Sundaram A. A novel method for pathogen reduction in wastewater treatment. J Environ Health Sci Eng 2013; 11: 12.
30. Iebba V, Santangelo F, Totion 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.
31. Yu T, Li G, Liu W, Hu H, Lu y. Coagulation increased the growth potential of various species bacteria of effluent of a MBR for the treatment of domestic wastewater. Environ Sci Pollut Res Int 2017; 24: 5126-5133.
How to Cite
Jafarian N, Akhavan Sepahi A, Naghavi NS, Hosseini F, Nowroozi J. Using autochthonous Bdellovibrio as a predatory bacterium for reduction of Gram-negative pathogenic bacteria in urban wastewater and reuse it. Iran J Microbiol. 12(6):556-564.
Original Article(s)