Isolation and Characterization of Novel Cocktail Phages of Multidrug-Resistant (MDR) Acinetobacter Baumannii

Main Article Content

Nkechi V. Enwuru
Jachike T. Nzekwe
Bisola O. Ojo
Abdul-Rahman Usman
Folake R. Peters


Background: Acinetobacter baumannii is a nosocomial organism, classied as a critical pathogen with multidrug-resistant (MDR) challenges thus, it is urgently necessary to develop novel antimicrobial strategies. Phage therapy is an alternative and promising strategy, a highly effective biocontrol agent against MDR A. baumannii. This study characterizes three phages of MDR A. baumannii.

Method: Acinetobacter baumannii was isolated from clinical wound specimens and was identied using VITEK® 2 system (Biomerieux, France). The antibiotic susceptibility prole was determined according to the Clinical and Laboratory Standard Institute (CLSI). Phages specic to MDR A. baumannii were isolated from sewage and irrigation channel samples, then tested for pH and temperature stability.

Results: Among the A. baumanniistrains isolated, ve strains (Ab 140, Ab 150,Ab 333, Ab 1289, and Ab 976) showed remarkable resistance to antibiotics tested, with the resistance rate ranging from 90 to 40 %, respectively. Three lytic phages specic to Ab 140, Ab 150, and Ab 333 were isolated among ve MDR A. bamannnii isolates used for phage isolation. Lytic Phages specic to Ab 140 and Ab 150 produced clear big plaques with halo-like appearances around the inhibition zone, while Ab 333 produced clear and small size plaques. The phages were designated TJ 140, TJ 150, and TJ 333 and were stable in various pH and temperatures with a high survival rate in pH of 5.0 to 7.0 and 20 to 60 oC, respectively.

Conclusion: The isolated phages exhibited strong lytic activity against MDR A. baumannii isolates tested and are stable in various pH and temperature ranges. They had no lytic eect on the heterogeneous strains and are good potential candidates for therapeutic applications.

Article Details

How to Cite
Enwuru, N. V., Nzekwe, J. T., Ojo, B. O., Usman, A.-R., & Peters, F. R. (2023). Isolation and Characterization of Novel Cocktail Phages of Multidrug-Resistant (MDR) Acinetobacter Baumannii: The Nigerian Journal of Pharmacy, 57(1), 435–446. Retrieved from


Dhingra S, Rahman NAA, Peile E, Rahman M, Sartelli M, Hassali MA, et al. 2020 Microbial Resistance Movements: An Overview of Global Public Health Threats Posed by Antimicrobial Resistance, and How Best to Counter. Front Public Health. 4;8:531.

Lai CC, Chen SY, Ko WC, and Hsueh PR. 2021 Increased antimicrobial resistance during the COVID-19 pandemic. Int J Antimicrob Agents. 57(4):106324.

Serra-Burriel M, Keys M, Campillo-Artero C, Agodi A, Barchitta M, Gikas A, et al. 2020 Impact of multi-drug resistant bacteria on economic and clinical outcomes of healthcare-associated infections in adults: Systematic review and meta-analysis. PLoS One 15(1): e0227139. Available from:

Shrivastava SR, Shrivastava PS, and Ramasamy J. 2018 World health organization releases global priority list of antibiotic-resistant bacteria to guide research, discovery, and development of new antibiotics. Journal of Medical Society 32(1):76. Available from:;year=2018;volume=32;issue=1;spage=76;epage=77;aulast=Shrivastava

Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet DL, et al. 2018 Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis. 1;18(3):318–27.

Centers for Disease control and Prevention. Antibiotic resistance threats in the United States, 2018. Atlanta, Georgia; 2019 Nov. Available from:

World Health Organization. Global Antimicrobial Resistance and Use Surveillance System (GLASS) Report. 2021. Available from:

Nasr P. 2020 Genetics, epidemiology, and clinical manifestations of multidrug-resistant Acinetobacter baumannii. J Hosp Infect. 104(1):4–11. Available from:

Moubareck CA and Halat DH. 2020 Insights into Acine to bacterbaumann ii: A Review of Microbiological, Virulence, and Resistance Traits in a

Threatening Nosocomial Pathogen. Antibiotics. 9(3):119. Available from:

Sarshar M, Behzadi P, Scribano D, Palamara AT, and Ambrosi C. 2021 baumannii: An Ancient Commensal with Weapons of a Pathogen. Pathogens 10(4) :387. Available from:

De Oliveira DMP, Forde BM, Kidd TJ, Harris PNA, Schembri MA, Beatson SA, et al. 2020 Antimicrobial resistance in ESKAPE pathogens. Clin

Microbiol Rev 33(3). Available from:

Ogbolu DO, Alli OAT, Oluremi AS, Ogunjimi YT, Ojebode DI, Dada V, et al. 2020 Contribution of NDM and OXA-type carbapenemases to carbapenem resistance in clinical Acinetobacter baumannii from Nigeria. Infectious Diseases 52(9):644–50.

Higgins PG, Dammhayn C, Hackel M, and Seifert H. 2010 Global spread of carbapenem-resistant Acinetobacter baumannii. Journal of Antimicrobial Chemotherapy. 65(2):233–8. Available from:

Nguyen M, and Joshi SG. 2021 Carbapenem resistance in Acinetobacter baumannii, and their importance in hospital-acquired infections: a scientific review. J Appl Microbiol. 131(6):2715–38. Available from:

Qureshi ZA, Hittle LE, O'Hara JA, Rivera JI, Syed A, Shields RK, et al. 2015 Colistin-Resistant Acinetobacter baumannii: Beyond Carbapenem

Resistance. Clinical Infectious Diseases 60(9): 1295 – 303. Available from:

Doi Y, Murray GL, and Peleg AY. 2015 Acinetobacter baumannii: evolution of antimicrobial resistance-treatment options. Semin Respir Crit Care Med 36(1): 85–9 8. Available from:

Hyman P. 2019 Phages for Phage Therapy: Isolation, Characterization, and Host Range Breadth. Pharmaceuticals 12(1):35. Available from:

Abedon ST, García P, Mullany P, and Aminov R. 2017 Editorial: Phage Therapy: Past, Present and Future. Front Microbiol 8:981. Available from:

Bagińska N, Pichlak A, Górski A, and JończykMa tysi ak E. 2019 Specific and Selective

Bacteriophages in the Fight against Multidrugresistant Acinetobacter baumannii. Virologica Sinica 34 (4): 347 – 57. Available from:

Kusradze I, Karumidze N, Rigvava S, Dvalidze T, Katsitadze M, Amiranashvili I, et al. 2016 Characterization and testing the efficiency of Acinetobacter baumannii phage vB-GEC_Ab-M-G7 as an antibacterial agent. Front Microbiol. 4; 7(OCT): 1590.

Grygor c ewi c z B, Ros z ak M, Gol e c P, Śleboda‐taront D, Łubowska N, Górska M, et al. 2020 Antibiotics Act with vB_AbaP_AGC01 Phage against Acinetobacter baumannii in Human Heat-Inactivated Plasma Blood and Galleria mellonella Models. International Journal of Molecular Sciences 21(12): 4390. Available from:

Wu N, Dai J, Guo M, Li J, Zhou X, Li F, et al. 2021 Pre-optimized phage therapy on secondary baumannii: In vitro and in vivo. Iran J Basic Med Sci. 1; 21(11): 1100–8.

Wintachai P, Surachat K, and Singkhamanan K. 2022 Isolation and Characterization of a Novel Autographiviridae Phage and Its Combined Effect with Tigecycline in Controlling Multidrug-Resistant Acinetobacter baumannii-Associated Skin and Soft Tissue Infections. Viruses. 1; 14(2).

Wintachai P, Naknaen A, Pomwised R, Voravuthikunchai SP, and Smith DR. 2019 Isolation and characterization of Siphoviridae phage infecting extensively drug-resistant Acinetobacter baumannii and evaluation of therapeutic efficacy in vitro and in vivo. J Med Microbiol 68(7):1096–108. Available from:

Jeon J, Park JH, and Yong D. Efficacy of bacteriophage treatment against carbapenem-resistant Acinetobacter baumannii in Galleria mellonella larvae and a mouse model of acute pneumonia. BMC Microbiol 19(1): 1 – 14. Available from:

Miller H. 1987 Practical aspects of preparing phage and plasmid DNA: growth, maintenance, and storage of bacteria and bacteriophage. Methods Enzymol [Internet]. 152(C): 145–70.

Alvi IA, Asif M, Tabassum R, Abbas Z, and Ur Rehman S. 2018 Storage of bacteriophages at 4°C leads to no loss in their titer after one year. Pak J Zool. Dec1; 50(6): 2395 – 8.