doi: 10.14202/vetworld.2017.973-978
Share this article on [Facebook] [LinkedIn]
Article history: Received: 26-04-2017, Accepted: 25-07-2017, Published online: 23-08-2017
Corresponding author: C. Sannat
E-mail: csannat@rediffmail.com
Citation: Shende RK, Hirpurkar SD, Sannat C, Rawat N, Pandey V (2017) Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens, Veterinary World, 10(8): 973-978.Aim: Present investigation was conducted to isolate and characterize bacteriophages with lytic activity against common bacterial pathogens.
Materials and Methods: A total of 60 samples of animal waste disposal from cattle (42) and buffalo (18) farms were collected from three different strata, i.e., top, mid, and bottom of collection tank. Samples were primarily subjected to rapid detection methods, and then isolation of phage was done by double agar layer method using Bacillus subtilis (BsH) and Escherichia coli (EH) as host system. Phages were characterized on the basis of plaque morphology, temperature, pH susceptibility, and host range.
Results: Recovery of phages was higher from dairy cattle farm waste (78.57%) as compared to buffalo farm waste (72.22%) and bottom layer of tank showed maximum recovery. Bacillus subtilis (91%) supported the growth of more phages as compared to E. coli (9%). Three different phage morphotypes were observed each against Bacillus subtilis (BsHR1, BsHR2, and BsHR3) and E. coli (EHR1, EHR2, and EHR3). Mean phage titer of above six phage isolates ranged between 3x1010 and 5x1012 plaque forming units/ml. Viability of phages was by, and large unaffected at 70°C within 2-3 min, and phage isolates were completely inactivated below pH 3 and above 11. Coliphage EHR1 had widest host range followed by BsHR1 and BsHR2 while EHR2, EHR3, and BsHR3 had low lytic activity.
Conclusion: It could be concluded from the present study that the Bacillus and Coli phage has wide host range and thus exhibits the potential to be used as drug substitute tool against common bacterial pathogens.
Keywords: Bacillus subtilis, bacteriophage, Escherichia coli, host range, sewage.
1. Zhan, A., Buchan, A. and Chen, F. (2015) Novel N4 bacteriophages prevail in the cold biosphere. Appl. Environ. Microbiol., 81: 5196-5202. [Crossref] [PubMed] [PMC]
2. Naghavi, N.S., Golgoljam, M. and Akbari, M. (2013) Effect of three sewage isolated bacteriophages on the multi drug resistant pathogenic bacteria. J. Biol. Sci., 13: 422-426. [Crossref]
3. WHO. (2014) Antimicrobial Resistance: Global Report on Surveillance. World Health Organization. Available from: http://www.who.int/drugresistance/documents/AMR_report_Web_slide_set.pdf?ua=1 Accessed on 09.08.2017.
4. Clarke, T. (2003) Drug companies snub antibiotics as pipeline threatens to run dry. Nature, 425: 225. [Crossref] [PubMed]
5. O'Flynn, G., Ross, R.P., Fitzgerald, G.F. and Coffey, A. (2004) Evaluation of a cocktail of three bacteriophages for biocontrol of Escherichia coli O157:H7. Appl. Environ. Microbiol., 70: 3417-3424. [Crossref] [PubMed] [PMC]
6. Holmfeldt, K., Middelboe, M., Nybroe, O. and Riemann, L. (2007) Large variabilities in host strain susceptibility and phage host range govern interactions between lytic marine phages and their Flavobacterium hosts. Appl. Environ. Microbiol., 73: 6730-6739. [Crossref]
7. Beke, G., Stano, M. and Klucar, L. (2016) Modelling the interaction between bacteriophages and their bacterial hosts. Math. Biosci., 279: 27-32. [Crossref] [PubMed]
8. Jothikumar, N., Reddy, C.G., Sundaria, R.B. and Saigopalb, D.V.R. (2000) Isolation of coliphages specific to enterotoxigenic E. coli (ETEC). J. Environ. Monit., 2: 372-374. [Crossref] [PubMed]
9. Harrigan, W.F. and McCance, M.E. (1993) Laboratory Methods in Food Dairy Microbiology. Academic Press, Harcourt Brace and Company Publishers, London, U.K. p52-57.
10. Adams, M.H. (1959) Discovery of bacteriophages and methods of study of bacterial viruses. Bacteriophages. Vol. 42. Interscience Publication, New York. p447-448.
11. Ellis, C.B. and Winters, A.L. (1969) Isolation of potential MS2 bacteriophage strains. Biol. Sci., 85: 336-345.
12. Tiwari, R., Hirpurkar, S.D. and Shakya, S. (2010) Isolation and characterization of lytic phage from natural waste material of livestock. Indian Vet. J., 87: 644-646.
13. Shukla, S. and Hirpurkar, S.D. (2011) Recovery status of bacteriophages of different livestock farms of Veterinary College, Adhartal, Jabalpur, India. Vet. World, 4(3): 117-119.
14. Verma, H., Pramod, D., Abbas, M., Prajapati, A., Ramchandra, D. and Rawat, M. (2013) Isolation and partial characterization of lytic phage against Salmonella abortusequi. Vet. World, 6(2): 72-75. [Crossref]
15. Mattila, S., Ruotsalainen, P. and Jalasvuori, M. (2015) On-demand isolation of bacteriophages against drug-resistant bacteria for personalized phage therapy. Front. Microbiol., 6: 1271. [Crossref] [PubMed] [PMC]
16. Yuan, Y., Gao, M., Wu, D., Liu, P. and Wu, Y. (2012) Genome characteristics of a novel phage from Bacillus thuringiensis showing high similarity with phage from Bacillus cereus. PLoS One, 7(5): 375-557. [Crossref]
17. Gillis, A. and Mahillon, J. (2014) Phages preying on Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis: Past, present and future. Viruses, 6: 2623-2672. [Crossref] [PubMed] [PMC]
18. Krasowska, A., Biegalska, A., Augustyniak, D., Los, M., Richert, M. and Lukaszewicz, M. (2015) Isolation and characterization of phages infecting Bacillus subtilis. Biomed. Res. Int., 2015: Article ID: 179597, 10.
19. Askora, A., Merwad, A., Gharibe, R.M. and Maysa, A.I.A. (2015) A lytic bacteriophage as a biocontrol for some enter pathogenic and enter hemorrhagic Escherichia coli strains of zoonotic risk in Egypt. Revue Med. Vet., 166(34): 76-83.
20. Carey-Smith, G.V., Billington, C., Cornelius, A.J., Hudson, J.A. and Heinemonn, J.A. (2006) Isolation and characterization bacteriophages and infecting Salmonella spp. FEMS Microb. Lett., 258: 182-186. [Crossref] [PubMed]
21. Goyal, S.M., Gerba, C.P. and Bitton, G. (1987) In: Phage Ecology. John Wiley and Sons, New York. p321.
22. Ghasemian, A., Bavand, M. and Moradpour, Z. (2017) A broad-host range coliphage against a clinically isolated E. coli O157: Isolation and characterization. J. Appl. Pharm. Sci., 7(3): 123-128.
23. Pedroso, D.M.M. and Martins, M.T. (1995) Ultra-morphology of coliphages isolated from water. Water Res., 29: 1199-1202. [Crossref]
24. Ibrahim, W.N.W., Aznan, A.S., Saari, N.A., Leong, L.K., Musa, N., Razzak, L.A., Danish-Daniel, M., Zainathan, S.C., Din, M.S.M., Ghaffar, M.A. and Musa, N. (2017) In-vitro characterization of lytic bacteriophage PhVh6 as potential biocontrol agent against pathogenic Vibrio harveyi. AACL Bioflux, 10(1): 64-76.
25. Langlet, J., Gaboriaud, F. and Gantzer, C. (2007) Effects of pH on plaque forming unit counts and aggregation of MS2 bacteriophage. J. Appl. Microbiol., 103: 1632-1638. [Crossref] [PubMed]
26. Lu, Z., Breidt, F., Fleming, H.P., Altermann, E. and Klaenhammer, T.R. (2003) Isolation and characterization of a Lactobacillus plantarum bacteriophage phiJL-1, from a cucumber fermentation. Int. J. Food Microbiol., 84: 225-235. [Crossref]
27. McLaughlin, M.R., Balaa, M.F., Sims, J. and King, R. (2006) Isolation of Salmonella Bacteriophages from swine effluent lagoons. J. Environ. Qual., 35: 522-528. [Crossref] [PubMed]
28. Bielke, H., Donoghue, S., Donoghue, A. and Hargis, D.B.M. (2007) Salmonella Host range of bacteriophges that infect multiple genera. Poult. Sci., 86: 2536-2540. [Crossref] [PubMed]
29. Ackermann, H.W. (2007) Phages examined in the electron microscope. Arch. Virol., 152: 227-243. [Crossref] [PubMed]
30. Ahmed, K., Kaderbhai, N.N. and Kaderbhai, M.A. (2012) Bacteriophage therapy revisited. Afr. J. Microbiol. Res., 6(14): 3366-3379.
31. Marwa, B.S. and Abdulamir, A.S. (2014) In vitro study on using bacteriophages in the treatment of pathogenic Escherichia coli in Iraq. Iraqi J. Med. Sci., 12(2): 128-132.
32. Morello, E., Emilie, S., Damien, M., Michel, H., Laurent, T. and Lhousseine, D. (2011) Pulmonary bacteriophage therapy on Pseudomonas aeruginosa cystic fibrosis strains first steps towards treatment and prevention. PLoS One, 10: 13-71. [Crossref]
33. Garcia, P., Madera, C., Martinez, B., Rodriguez, A. and Suarez, J.E. (2009) Prevalence of bacteriophages infecting Staphylococcus aureus in dairy samples and their potential as biocontrol agents. J. Dairy Sci., 92: 3019-3026. [Crossref] [PubMed]