Abstract
Escherichia coli has been enlisted as a pathogen of antimicrobial resistance (AMR) importance posing a threat to human health. In the wake of the widespread AMR, lytic bacteriophages remain a promising alternative to antibiotics for controlling the multidrug-resistant (MDR) pathogens. However, the major hindrance for phage therapy is the lack of a rapid procedure to harness broad host range phages. Hitherto in vogue, lytic phage isolation methods are based on conventional single host enrichment (M1). Limitation to the available methods includes a higher number of steps, additional instrumentation, cost and time involved in the screening of samples and more recovery of narrow host range phages.To improve the conventional method of isolation, a multiple host enrichment approach (modified method; M2) is developed and validated with a classical approach using 24 MDR extended-spectrum β-lactamase producing (ESBL) E. coli and universal coliphage host E. coli NCIM 2089. A total of 58 phages were isolated using both the methods (27 phages from M1 and 31 phages from M2) and were categorized, based on the DNA restriction digestion pattern, to 25 groups with a maximum host range of 17 in host range analysis. Even though there is no significant difference (P > 0.01) in recovering number of phages between M1 and M2 methods, excepting in one occasion where MFB13 host yielded 3 more morphologically distinct phages in M2 (p ≤ 0.05), the M2 approach was better in terms of reduction in time (4.72%) and cost (23.5%) and same instrumentation requirement. This emphasizes that the proposed method has the potential to yield broad host range lytic phages against localized strain repository of AMR E. coli which can be leading to its application in food, food contact surfaces as well as phage therapy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Data generated from the study is submitted as Supplementary Table S1.
Abbreviations
- MFB:
-
Microbiology Fermentation and Biotechnology Division
- NCIM:
-
National Collection of Industrial Microorganisms
- CLSI:
-
Clinical & Laboratory Standards Institute
- ATCC:
-
American Type Culture Collection
- PES:
-
Polyethersulfone
- SPSS:
-
Statistical Package for the Social Sciences
References
Abedon ST, Kuhl SJ, Blasdel BG, Kutter EM (2011) Phage treatment of human infections. Bacteriophage 1(2):66–85. https://doi.org/10.4161/bact.1.2.15845
Adams MH (1959) Host specificity. In: Benzer S et al. (ed) Bacteriophages. Interscience Publishers Ltd, London, pp 121–135
Akhwale JK, Rohde M, Rohde C et al (2019) Isolation, characterization and analysis of bacteriophages from the haloalkaline lake Elmenteita, Kenya. PLoS One 14(4):e0215734. https://doi.org/10.1371/journal.pone.0215734
Bhatia R (2019) Implementation framework for One Health approach. Indian J Med Res 149(3):329. https://doi.org/10.1002/ange.19620740437
Bren L (2007) Bacteria-eating virus approved as food additive. FDA Consum 41(1):20–22
Carlson K (2005) Working with bacteriophages: Common techniques and methodological approaches. In: Kutter E, Sulakvelidze A (eds) Bacteriophages: Biology and Application. CRC Press, Boca Raton, pp 437–494
Ceyssens PJ, Glonti T, Kropinski NM et al (2011) Phenotypic and genotypic variations within a single bacteriophage species. Virol J 8:134. doi:https://doi.org/10.1186/1743-422X-8-134 Published 2011 Mar 23.
Chan BK, Abedon ST, Loc-Carrillo C (2013) Phage cocktails and the future of phage therapy. Future microbiol 8(6):769 –769 83. https://doi.org/10.2217/fmb.13.47
Chanishvili N (2012) Phage therapy–history from Twort and d’Herelle through Soviet experience to current approaches. Adv Virus Res 83:3–40. https://doi.org/10.1016/B978-0-12-394438-2.00001-3
Chhibber S, Kaur P, Gondil VS (2018) Simple drop cast method for enumeration of bacteriophages. J Virol Methods 262:1–5. https://doi.org/10.1016/j.jviromet.2018.09.001
CLSI (2018) Performance standards for antimicrobial disk susceptibility tests. CLSI document M02-A11 32(1), Wayne
Diaz-Munoz SL, Koskella B (2014) Bacteria–phage interactions in natural environments. In: Sima S, Geoffrey G (eds) Advances in applied microbiology, 1st edn. Academic, Michael, pp 135–183
Drulis-Kawa Z, Majkowska-Skrobek G, Maciejewska B, Delattre AS, Lavigne R (2012) Learning from bacteriophages-advantages and limitations of phage and phage-encoded protein applications. Curr Protein PeptSci 13(8):699–722. https://doi.org/10.2174/138920312804871193
Eydal HS, Jägevall S, Hermansson M, Pedersen K (2009) Bacteriophage lytic to Desulfovibrioaespoeensis isolated from deep groundwater. The ISME J 3(10):1139–1147. https://doi.org/10.1038/ismej.2009.66
Fister S, Robben C, Witte AK, Schoder D, Wagner M, Rossmanith P (2016) Influence of environmental factors on phage–bacteria interaction and on the efficacy and infectivity of phage P100. Front Microbiol 7:1152. https://doi.org/10.3389/fmicb.2016.01152
Gelman D, Eisenkraft A, Chanishvili N, Nachman D, Coppenhagem Glazer S, Hazan R (2018) The history and promising future of phage therapy in the military service. J Trauma Acute Care Surg 85:S18–S26. https://doi.org/10.1097/TA.0000000000001809
Hyman P, Abedon ST (2010) Bacteriophage host range and bacterial resistance. Adv Appl Microbiol 70:217–248. https://doi.org/10.1016/S0065-2164(10)70007-1
Jassim SA, Limoges RG (2014) Natural solution to antibiotic resistance: bacteriophages ‘The Living Drugs’. World J Microbiol Biotechnol 30(8):2153–2170. https://doi.org/10.1007/s11274-014-1655-7
Jensen EC, Schrader HS, Rieland B, Thompson TL, Lee KW, Nickerson KW, Kokjohn TA (1998) Prevalence of broad-host-range lytic bacteriophages of Sphaerotilus natans, Escherichia coli, and Pseudomonas aeruginosa. Appl Environ Microbiol 64(2):575–80. https://doi.org/10.1128/AEM.64.2.575-580.1998
Kim S, Schuler B, Terekhov A, Auer J, Mauer LJ, Perry L, Applegate B (2009) A bioluminescence-based assay for enumeration of lytic bacteriophage. J Microbiol Methods 79(1):18–22. https://doi.org/10.1016/j.mimet.2009.07.011
Kirby WM, Bauer AW, Sherris JC. turck, Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45(4):493. https://doi.org/10.1093/ajcp/45.4_ts.493
Knezevic P, Petrovic O (2008) A colorimetric microtiter plate method for assessment of phage effect on Pseudomonas aeruginosa biofilm. J Microbiol Methods 74(2–3):114–8. https://doi.org/10.1016/j.mimet.2008.03.005
Kutateladze M, Adamia R (2010) Bacteriophages as potential new therapeutics to replace or supplement antibiotics. Trends Biotechnol 28(12):591–595. https://doi.org/10.1016/j.tibtech.2010.08.001
Kutter E (2009) Phage host range and efficiency of plating. Methods MolBiol 501:141–149
Lin L, Han J, Ji X, Hong W, Huang L, Wei Y (2011) Isolation and characterization of a new bacteriophage MMP17 from Meiothermus. Extremophiles 15(2):253–258. https://doi.org/10.1007/s00792-010-0354-z
Lin DM, Koskella B, Lin HC (2017) Phage therapy: An alternative to antibiotics in the age of multi-drug resistance. World J Pharmacol 8(3):162. https://doi.org/10.4292/wjgpt.v8.i3.162
Liu W, Li C, Qiu ZG, Jin M, Wang JF, Yang D, Xiao ZH, Yuan ZK, Li JW, Xu QY, Shen ZQ (2017) Development of a novel and highly efficient method of isolating bacteriophages from water. J Microbiol Methods 139:143–149. https://doi.org/10.1016/j.mimet.2017.05.019
Lu TK, Koeris MS (2011) The next generation of bacteriophage therapy. Curr Opin Microbiol 14(5):524–531. https://doi.org/10.1016/j.mib.2011.07.028
Lu Z, BreidtJr F, Fleming HP, Altermann E, Klaenhammer TR (2003) Isolation and characterization of a Lactobacillus plantarum bacteriophage, ΦJL-1, from a cucumber fermentation. Int J Food Microbiol 84(2):225–235. https://doi.org/10.1016/s0168-1605(03)00111-9
Lu L, Cai L, Jiao N, Zhang R (2017) Isolation and characterization of the first phage infecting ecologically important marine bacteria Erythrobacter. Virol J 14(1):104. https://doi.org/10.1186/s12985-017-0773-x
Mancos M, Sramkova Z, Peterkova D, Vidova B, Godany A (2020) Functional expression and purification of tailor-made chimeric endolysin with the broad antibacterial spectrum. Biologia. https://doi.org/10.2478/s11756-020-00508-9
Mann HB, Whitney DR (1947) On a test of whether one of two random variables is stochastically larger than the other. Ann Math Stat 1:50–60. https://doi.org/10.1214/aoms/1177730491
Mapes AC, Trautner BW, Liao KS, Ramig RF (2016) Development of expanded host range phage active on biofilms of multi-drug resistant Pseudomonas aeruginosa. Bacteriophage 6(1):e1096995. https://doi.org/10.1080/21597081.2015.1096995
Mathur MD, Vidhani S, Mehndiratta PL, Bhalla P, Reddy BS (2003) Bacteriophage therapy: an alternative to conventional antibiotics. J Assoc Physicians India 51:593–596. https://doi.org/10.1258/095646202760159701
McLaughlin MR (2007) Simple colorimetric microplate test of phage lysis in Salmonella enterica. J Microbiol Methods 69(2):394–398. https://doi.org/10.1016/j.mimet.2007.01.006
McLaughlin MR, Balaa MF, Sims J, King R (2006) Isolation of Salmonella bacteriophages from swine effluent lagoons. J Environ Qual 35(2):522–528. https://doi.org/10.2134/jeq2005.0080
Melo LDR, Veiga P, Cerca N, Kropinski AM, Almeida C, Azeredo J, Sillankorva S (2016) Development of a Phage Cocktail to Control Proteus mirabilis Catheter-associated Urinary Tract Infections. Frontiers in Microbiology 7
Moon K, Kang I, Kim S, Kim SJ, Cho JC (2018) Genomic and ecological study of two distinctive freshwater bacteriophages infecting a Comamonadaceae bacterium. Sci Rep 8(1):1–9. https://doi.org/10.1038/s41598-018-26363-y
Morozova VV, Vlassov VV, Tikunova NV (2018) Applications of bacteriophages in the treatment of localized infections in humans. Front Microbiol9: 1696.https://doi.org/10.3389/fmicb.2018.01696
Myelnikov D (2018) An alternative cure: the adoption and survival of bacteriophage therapy in the USSR, 1922–1955. J Hist Med All Sci 73(4):385–411. https://doi.org/10.1093/jhmas/jry024
Nobrega FL, Costa AR, Santos JF, Siliakus MF, Van Lent JW, Kengen SW, Azeredo J, Kluskens LD (2016) Genetically manipulated phages with improved pH resistance for oral administration in veterinary medicine. Sci Rep 6:39235. https://doi.org/10.1038/srep39235
Rao BM, Lalitha KV (2015) Bacteriophages for aquaculture: are they beneficial or inimical. Aquaculture 437:146–154. https://doi.org/10.1016/j.aquaculture.2014.11.039
Rao BM, Surendran PK (2003) Coliphage test: a quick and easy method to detect faecal pollution in water and Fish. In: Surendran PK, Mathew PT, Thampuran N, Nambiar VN, Joseph J, Boopendranath MR, Lakshmanan PT, Nair PGV (eds) Seafood Safety. Society of Fisheries Technologists (India), Cochin, pp 556–561
Richards GP (2014) Bacteriophage remediation of bacterial pathogens in aquaculture: a review of the technology. Bacteriophage 4(4):e975540. https://doi.org/10.4161/21597081.2014.975540
Ross A, Ward S, Hyman P (2016) More is better: selecting for broad host range bacteriophages. Front Microbiol 7:1352. https://doi.org/10.3389/fmicb.2016.01352
Sarker SA, McCallin S, Barretto C, Berger B, Pittet AC, Sultana S, Krause L, Huq S, Bibiloni R, Bruttin A, Reuteler G (2012) Oral T4-like phage cocktail application to healthy adult volunteers from Bangladesh. Virology 434(2):222 –222 32. https://doi.org/10.1016/j.virol.2012.09.002
Shende RK, Hirpurkar SD, Sannat C, Rawat N, Pandey V (2017) Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens. Vet world 10(8):973. https://doi.org/10.14202/vetworld.2017.973-978
Son HM, Duc HM, Masuda Y et al (2018) Application of bacteriophages in simultaneously controlling Escherichia coli O157:H7 and extended-spectrum beta-lactamase producing Escherichia coli. Appl Microbiol Biotechnol 102:10259–10271. https://doi.org/10.1007/s00253-018-9399-1
Sulakvelidze A, Alavidze Z, Morris JG (2001) Bacteriophage therapy. Antimicrob Agents Ch 45(3):649–659. https://doi.org/10.1128/AAC.45.3.649-659.2001
Sundar MM, Nagananda GS, Das A, Bhattacharya S, Suryan S (2009) Isolation of host-specific bacteriophages from sewage against human pathogens. Asian J Biotechnol 1(4):163 –163 70. https://doi.org/10.3923/ajbkr.2009.163.170
Sybesma W, Zbinden R, Chanishvili N, Kutateladze M, Chkhotua A, Ujmajuridze A, Mehnert U, Kessler TM (2016) Bacteriophages as potential treatment for urinary tract infections. Front Microbiol 7:465. https://doi.org/10.3389/fmicb.2016.00465
Tiwari R, Dhama K, Kumar A, Rahal A, Kapoor S (2014) Bacteriophage therapy for safeguarding animal and human health: A review. Pak J Biol Sci 17:301–315. https://doi.org/10.3923/pjbs.2014.301.315
Van Twest R, Kropinski AM (2009) Bacteriophage enrichment from water and soil. Methods Mol Biol 501:15–21
VidovaB, Sramkova Z, Tisakova L, Oravkinova M, Godany A (2014) Bioinformatics analysis of bacteriophage and prophage endolysin domains. Biologia 69(5):541–556. https://doi.org/10.2478/s11756-014-0358-8
Withey S, Cartmell E, Avery LM, Stephenson T (2005) Bacteriophages—potential for application in wastewater treatment processes. Sci Total Environ 339(1–3):1–8. https://doi.org/10.1016/j.scitotenv.2004.09.021
World Health Organization (2015) Global action plan on antimicrobial resistance. Retrieved from https://apps.who.int/iris/handle/10665/193736. Accessed 3 Oct 2020
World Health Organization (2017) WHO publishes list of bacteria for which new antibiotics are urgently needed. Retrieved from https://www.who.int/news/item/27-02-2017-who-publishes-list-of-bacteria-for-which-new-antibiotics-are-urgently-needed. Accessed 3 Oct 2020
Acknowledgements
Authors are grateful to the Department of Biotechnology, Government of India for funding this research (Grant no: BT/PR26949/AAQ/2/888/2017). The authors are also thankful to ICAR-Central Institute of Fisheries Technology for providing the support.
Funding
The project is funded by Department of Biotechnology, Government of India (Grant no: BT/PR26949/AAQ/2/888/2017).
Author information
Authors and Affiliations
Contributions
Murugadas Vaiyapuri, Mukteswar Prasad Mothadaka, and Madhusudana Rao Badireddy conceptualized the study; Murugadas Vaiyapuri, Karthika Raveendran, Iris George, Devika Gundubilli, Sivaraman Gopala Krishnan and Madhusudana Rao Badireddy performed the experiments, Murugadas Vaiyapuri, Karthika Raveendran, Madhusudana Rao Badireddy drafted the manuscript; Visnuvinayagam Sivam and Joshy Chalil George analysed the data; Mukteswar Prasad Mothadaka and Ravishankar Chandragiri Nagarajarao, reviewed and edited the manuscript.
Corresponding author
Ethics declarations
Conflicts of interest
There are no conflicts of interest with authors in publishing the research article.
Ethics approval
Laboratory animals were not used in the experiments.
Consent to participate
Not applicable.
Consent for publication
All the authors consented to publish the work.
Code availability
Not applicable.
Electronic supplementary material
ESM 1
(DOCX )
Rights and permissions
About this article
Cite this article
Vaiyapuri, M., Raveendran, K., George, I. et al. Comparison of single and multi-host enrichment approach for harnessing lytic phages against antimicrobial-resistant E. coli: Repurposing the enrichment step. Biologia 76, 1041–1052 (2021). https://doi.org/10.2478/s11756-020-00652-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.2478/s11756-020-00652-2