Biological Characteristics and Application of Enteroinvasive Escherichia coli Phage DK-13

doi:10.13560/j.cnki.biotech.bull.1985.2023-0902

Abstract

Abstract:

【Objective】 Selection and isolation of phage that can lyse Enteroinvasive Escherichia coli (EIEC), analyzing its biological characterization, and investigating its sterilizing effect on contaminated pork. 【Method】 The double-layer agar method was used to isolate phages, and then the optimal multiplicity of infection，one-step growth curve and other biological characteristics of the isolate were determined. The genetic genomes were sequenced and analyzed, and the lysis efficiency was evaluated.【Result】 A bacteriophage that specifically lysed EIEC was isolated from hospital sewage and named DK-13. It has a typical tadpole-like morphology, consisting of a head and a contractile helical symmetric tail. It belongs to the family Myoviridae. The biological characterization showed that the optimal multiplicity of infection was 0.01, the incubation period was about 10 min, the lysis period was about 70 min, and the phage DK-13 was able to survive at 50℃ and pH 5.0-10.0. The whole genome sequencing showed that the phage genome was about 172 275 bp in length, with a GC content of 40.18%, and was predicted to have a total of 293 open reading frames（ORFs）, and no resistance genes or virulence genes were found. the sterilization effect in the contaminated pork was good, and the number of host bacteria significantly reduced. 【Conclusion】 A new virile phage DK-13 was isolated and identified, which has the advantages of short incubation period and high cleavage efficiency, and has great potential for application in food safety.

Key words: Enteroinvasive Escherichia coli, phage, phage application, food safety

WANG Meng-ya, LIU Jia-qi, JIANG Hai-lin, LI Jing-hua, ZHAO Chun-yan, HUANG Hong-lan. Biological Characteristics and Application of Enteroinvasive Escherichia coli Phage DK-13[J]. Biotechnology Bulletin, 2024, 40(3): 296-304.

Figures/Tables 7

Fig. 1 Morphological characteristics of phage DK-13 A: Morphology of phage DK-13. B: Transmission electron micrograph of phage DK-13

Fig. 2 Biological characters of phage DK-13 A: MOI of phage DK-13. B: Step growth curve of phage DK-13. Different lower letters indicate significant difference among each group (P<0.05). The same below

Fig. 3 Biological characteristics of phage DK-13 A: Temperature resistance of phage DK-13. B: pH resistance of phage DK-13

Table 1 Host range of DK-13 phage

菌株名称 Strain name	菌株类型 Strain type	噬菌体DK-13的裂解性 Phage DK-13 lytic ability
大肠埃希菌A1 Escherichia coli A1	O157:H7	-
大肠埃希菌A2 E. coli A2	EPEC	-
大肠埃希菌A3 E. coli A3	O26:H11	-
大肠埃希菌A4 E. coli A4	O26:H11	-
大肠埃希菌A5 E. coli A5	O26:H11	-
大肠埃希菌A6 E. coli A6	O26:H11	-
大肠埃希菌A7 E. coli A7	O26:H11	-
大肠埃希菌A8 E. coli A8	O46:H38	-
大肠埃希菌A9 E. coli A9	O11:NM	-
大肠埃希菌A10 E. coli A10	O157:H7	-
大肠埃希菌A11 E. coli A11	O157:H7	-
大肠埃希菌A12 E. coli A12	O157:H7	-
大肠埃希菌A13 E. coli A13	O157:H7	-
大肠埃希菌A14 E. coli A14	O157:H7	-
大肠埃希菌A15 E. coli A15	O157:H7	-
蜡样芽孢杆菌L1 Bacillus cereus L1	*	-
蜡样芽孢杆菌L2 B. cereus L2	*	-
金黄色葡萄球菌B1 Streptococcus aureus B1	*	-
金黄色葡萄球菌B2 S. aureus B2	*	-
金黄色葡萄球菌B3 S. aureus B3	*	-
大肠埃希菌（宿主菌）D1 E. coli D1	EIEC	+

Fig. 4 Annotation diagram of the genome of DK-13 phage The direction of arrow indicates the direction of gene. Red: The structural gene module. Brown: The packaged genetic module. Blue: The nucleotide metabolism gene module. Orange: Duplication/ Recombination gene module. Pink: Transcription/ Translation gene module. Green: Other function module. Grey: Hypothetical protein. Black: GC content

Fig. 5 Phylogenetic analysis of phage DK-13

Fig. 6 Sterilization effect of phage DK-13 at different temperatures on contaminated pork A: Sterilization effect of phage DK-13 at 4℃. B: Sterilization effect of phage DK-13 at 37℃

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