Method for bacteriophage isolation against targetCampylobacterstrains

Abstract

Aims: Poultry meat is considered a major source of Campylobacter. This micro‐aerobic bacterium is commonly responsible for foodborne illness. This work focuses on the isolation of Campylobacter coli lytic bacteriophages (phages) against target C. coli strains. Methods and Results: A method involving the enrichment of free‐range chicken samples in a broth containing the target C. coli strains and salts (CaCl2 and MgSO4) was used for phage isolation. This method allowed the isolation of 43 phages that were active against 83% of the C. coli strains used in the isolation procedure. Approximately 65% of the phages were also effective against Campylobacter jejuni strains. Conclusions: The use of target pathogens in the phage isolation step improves the likelihood of detecting and isolating phages for the control of these specific strains. Significance and Impact of the Study: This technique will be valuable in the context of phage therapy for enriching for phages that are active against specifically identified strains of bacteria, for example from a food poisoning outbreak or epidemic strains resistant to multiple antibiotics. In these situations, using the conventional methods for searching for bacteriophages active for these particular strains can be a time‐consuming, if not an unsuccessful process. Using the isolation method described in this manuscript, the particular strains can be added to the enrichment broth increasing the probability of finding phages against them. Therefore, it will shorten the time needed for seeking phages able to lyse target strains, which in most of the cases, because of the rapid increase in antimicrobial‐resistant bacteria, is of crucial importance.

Characterization and Application of Lytic Bacteriophages against <i>Campylobacter</i> <i>jejuni</i> Isolated from Poultry in Japan

“…Of these, isolates PHC 10, 19, and 25, which have broad lytic spectra, also have similar head diameters, as reported in the literature for the majority of Campylobacter phages Atterbury et al, 2003b;Carvalho et al, 2010b;Hansen et al, 2007;Hwang et al, 2009;Loc Carrillo et al, 2005;Owens et al, 2013;Sørensen et al, 2015 . The remaining phage isolates, PHC5 and PHC22, have narrow lytic spectra and slightly smaller head diameters, being similar to those reported by Bigwood et al 2008 .…”

“…In some reports, Campylobacter phages have been isolated from chicken samples meat samples, intestine samples, fecal contents, and excreta Atterbury et al, 2003b;Carvalho et al, 2010b;Connerton et al, 2004;Firlieyanti et al, 2016;Hansen et al, 2007;Hwang et al, 2009;Janež et al, 2014;Owens et al, 2013 . To date, however, there are no reports on the isolation of bacteriophages against Campylobacter in Japan.…”

Isolation and Characterization of Listeria phages for Control of Growth of Listeria monocytogenes in Milk

“…Then, the EP tubes were centrifuged at 10,000 g for 10 min at 4°C and the supernatant was propagated by the double-layer method as described previously. Propagated bacteriophages were collected with 4 mL of SM buffer (Carvalho et al , 2010; Janež and Loc-Carrillo, 2013; Salama et al , 1989). Bacteriophages titer was determined by the soft agar overlay method.…”

Tackling Campylobacter: a Review

“…After the isolating step, phages have been characterized and special attention has been given to the lytic spectrum of each phage or a given mixture of phages. Globally, the sensitivity to individual bacteriophages varies according to the strain and specie (Bigwood et al, 2008;Carvalho et al, 2009;El-Shibiny et al, 2009;Loc-Carrillo et al, 2011;Carvalho et al, 2012b;Owens et al, 2013). To date, six Campylobacter phage sequences have been published and are available in EMBL and GeneBank (Timms et al, 2010;Kropinski et al, 2011;Carvalho et al, 2012a;Hammerl et al, 2012).…”