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. 2024 May 16;24(1):497.
doi: 10.1186/s12879-024-09402-0.

In vitro evaluation of two novel Escherichia bacteriophages against multiple drug resistant avian pathogenic Escherichia coli

Mobina Karami  1 , Ali Goudarztalejerdi  2 , Abdolmajid Mohammadzadeh  1 , Enayat Berizi  3
Affiliations

In vitro evaluation of two novel Escherichia bacteriophages against multiple drug resistant avian pathogenic Escherichia coli

Mobina Karami et al. BMC Infect Dis. .

Abstract

Background: In recent years, there has been a growing interest in phage therapy as an effective therapeutic tool against colibacillosis caused by avian pathogenic Escherichia coli (APEC) which resulted from the increasing number of multidrug resistant (MDR) APEC strains.

Methods: In the present study, we reported the characterization of a new lytic bacteriophage (Escherichia phage AG- MK-2022. Basu) isolated from poultry slaughterhouse wastewater. In addition, the in vitro bacteriolytic activity of the newly isolated phage (Escherichia phage AG- MK-2022. Basu) and the Escherichia phage VaT-2019a isolate PE17 (GenBank: MK353636.1) were assessed against MDR- APEC strains (n = 100) isolated from broiler chickens with clinical signs of colibacillosis.

Results: Escherichia phage AG- MK-2022. Basu belongs to the Myoviridae family and exhibits a broad host range. Furthermore, the phage showed stability under a wide range of temperatures, pH values and different concentrations of NaCl. Genome analysis of the Escherichia phage AG- MK-2022. Basu revealed that the phage possesses no antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and any E. coli virulence associated genes. In vitro bacterial challenge tests demonstrated that two phages, the Escherichia phage VaT-2019a isolate PE17 and the Escherichia phage AG- MK-2022. Basu exhibited high bactericidal activity against APEC strains and lysed 95% of the tested APEC strains.

Conclusions: The current study findings indicate that both phages could be suggested as safe biocontrol agents and alternatives to antibiotics for controlling MDR-APEC strains isolated from broilers.

Keywords: Antibiotic; Avian pathogenic Escherichia coli (APEC); Bacteriophage; Broilers; Colibacillosis; Multidrug resistant (MDR).

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
The plaques of Escherichia phage AG- MK-2022.Basu formed on a double-layered agar plate. A: 109PFU/mL. B: 103 PFU/mL
Fig. 2
Fig. 2
Transmission electron microscopy (TEM) of the Escherichia phage AG- MK-2022.Basu. A: Scale bar, 100 nm (Magnification = X50000) and B: Scale bar, 60 nm (Magnification = X85000)
Fig. 3
Fig. 3
Escherichia phage AG- MK-2022. Basu stability in various conditions: (A) temperature; (B) pH; (C) NaCl
Fig. 4
Fig. 4
Adsorption of Escherichia phage AG- MK-2022. Basu to E. coli ATCC 25,922 (A); One-step growth curves of Escherichia phage AG- MK-2022. Basu in the presence of E. coli ATCC 25,922 as host (B)
Fig. 5
Fig. 5
(a) RAPD band patterns obtained from Escherichia phage VaT-2019a isolate PE17(B1 and B2) and, Escherichia phage AG- MK-2022.Basu (K1 and K2) using P1 and P2 primers, respectively; (b) and (c) PCR for detection of Escherichia coli virulence associated genes stx1, stx2, and hylA. (L) marker 100 bp, (N) negative control, (P) positive control (Escherichia coli O157: H7 ATCC 43,895), (B1) Escherichia phage VaT-2019a isolate PE17 with primer P1, (K1) Escherichia phage AG- MK-2022.Basu with primer P1, (B2) Escherichia phage VaT-2019a isolate PE17 with primer P2, (K2) Escherichia phage AG- MK-2022.Basu with primer P2
Fig. 6
Fig. 6
Bacterial challenge test of phage Escherichia phage AG- MK-2022. with Escherichia. coli ATCC 25,922. Bac + phage: bacteria + phage, bac: bacteria
Fig. 7
Fig. 7
Bacteriolytic activity of Escherichia phage AG- MK-2022. Basu and, Escherichia phage VaT-2019a isolate PE17 against Avian Pathogenic Escherichia coli (APEC) strains in vitro
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