Bacteriophages Isolated from Chicken Meat and the Horizontal Transfer of Antimicrobial Resistance Genes

DOI: 10.1128/AEM.00872-15

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FIG 1
Proportion of resistance-transducing phages among all isolated phages. Kanamycin resistance-transducing phages were detected the most often (n = 43 of 243 phages detected), followed by chloramphenicol (n = 8) and tetracycline (n = 4) resistance-transducing phages. Three phages were found to transduce kanamycin plus ampicillin resistance, and two phages transduced kanamycin plus chloramphenicol resistance. ϕCm^r, ϕKm^r, ϕTet^r, ϕKm^r Amp^r, and ϕKm^r Cm^r, chloramphenicol, kanamycin, tetracycline, kanamycin and ampicillin, and kanamycin and chloramphenicol resistance-transducing phages, respectively.

TABLE 1

Primers and conditions for PCR of resistance genes

Target gene	Primer name	Sequence	Annealing temp (°C)	PCR amplification product size (bp)	Reference
tet(A)	tet(A)-f	5′-GCTACATCCTGCTTGCCTTC-3′	57	210	44
	tet(A)-r	5′-CATAGATCGCCGTGAAGAGG-3′
tet(B)	tet(B)-f	5′-TTGGTTAGGGGCAAGTTTTG-3′	57	659	44
	tet(B)-r	5′-GTAATGGGCCAATAACACCG-3′
tet(C)	tet(C)-f	5′-CTTGAGAGCCTTCAACCCAG-3′	55	418	44
	tet(C)-r	5′-ATGGTCGTCATCTACCTGCC-3′
aphA1	aphA-1a-f	5′-AACGTCTTGCTCGAGGCCGCG-3′	65	670	45
	aphA-1a-r	5′-GGCAAGATCCTGGTATCGGTCTGC-3′
bla_CMY	bla_CMY-f	5′-TGGCCGTTGCCGTTATCTAC-3′	55	870	46
	bla_CMY-r	5′-CCCGTTTTATGCACCCATGA-3′
bla_IMP	bla_IMP-f	5′-GAATAGAGTGGATTAATTCTC-3′	55	232	47
	bla_IMP-r	5′-GGTTTAAYAAAACAACCACC-3′
bla_OXA	bla_OXA-2-f	5′-CAAGCCAAAGGCACGATAGTTG-3′	56	561	47
	bla_OXA-2-r	5′-CTCAACCCATCCTACCCACC-3′
bla_PSE	bla_PSE-f	5′-TGCTTCGCAACTATGACTAC-3′	55	438	46
	bla_PSE-r	5′-AGCCTGTGTTTGAGCTAGAT-3′
bla_TEM-1	bla_TEM-1-f	5′-CAGCGGTAAGATCCTTGAGA-3′	55	643	46
	bla_TEM-1-r	5′-ACTCCCCGTCGTGTAGATAA-3′
catI	catI-f	5′-AGTTGCTCAATGTACCTATAACC-3′	50	547	48
	catI-r	5′-TTGTAATTCATTAAGCATTCTGCC-3′
floR	floR-f	5′-CGCCGTCATTCCTCACCTTC-3′	50	215	48
	floR-r	5′-GATCACGGGCCACGCTGTGTC-3′

TABLE 2
Numbers of chicken meat samples by type of resistant E. coli isolate
a Amp^r, Cm^r, Km^r, and Tet^r, ampicillin, chloramphenicol, kanamycin, and tetracycline resistant, respectively.

TABLE 3

Lysotypes of antimicrobial resistance-transducing phages

LT^a pattern	Lysis of the indicated E. coli strains					Phage(s) transducing resistance to the indicated antimicrobial^b
LT^a pattern	ATCC 13706	JM109	W3110	MC1061	DH5α	Kanamycin	Chloramphenicol	Ampicillin	Tetracycline
LT1	+++	+++	+++	+++	−	ϕE2b, ϕE5a, ϕE5b, ϕE5c, ϕE5d, ϕE5e	ϕE6b, ϕE16e, ϕE30e	ϕE5c, ϕE9b	ϕE10c, ϕE10d
						ϕE7a, ϕE7b, ϕE7c, ϕE7d, ϕE7e, ϕE9b	ϕE49d	ϕE13d	ϕE15d, ϕE40e
						ϕE9d, ϕE12b, ϕE12e, ϕE13a, ϕE13b
						ϕE13c, ϕE13d, ϕE13e, ϕE16a, ϕE16d
						ϕE19b, ϕE19c, ϕE22c, ϕE23b, ϕE23c
						ϕE28a, ϕE28b, ϕE28c, ϕE28d, ϕE30a, ϕE49a
LT2	+++	++	+	+++	−	ϕE2c	ϕE43e
LT3	+++	++	++	++	−	ϕE2d
LT4	+++	+	++	++	−	ϕE2e
LT5	+++	+	+++	++	−	ϕE6a, ϕE19a
LT6	+++	+	+++	+++	−	ϕE6d, ϕE9c, ϕE27c
LT7	+++	+	+	++	−	ϕE9a, ϕE15c
LT8	+++	+++	+	+++	−	ϕE12c, ϕE12d, ϕE34e	ϕE12c
LT9	+++	+	++	+	−	ϕE19e
LT10	+++	++	++	+++	−	ϕE34a	ϕE34a
LT11	+++	+	+	+++	−		ϕE35d
LT12	+++	++	+++	+++	−		ϕE35e
LT13	+++	−	−	−	−		ϕE39d

↵a LT, lysotype.
↵b The phage designations indicate the chicken meat sample (E) number (indicated by Arabic numerals) and the phage isolate number (indicated by lowercase letters); for example, ϕE5a represents the first phage isolate from chicken meat sample number 5.