1株跨属感染猪霍乱沙门菌和大肠杆菌烈性噬菌体的分离及其生物学特性

doi:10.11843/j.issn.0366-6964.2020.08.030

Abstract

Abstract: This experiment was conducted to isolate and characterize a lytic bacteriophage against Salmonella choleraesuis (S. choleraesuis) from the healthy pig gut. Chyme and mucosal samples from pig jejunum and colon were assayed for the presence of bacteriophage capable of forming plaques on S. choleraesuis 13122. Bacteriophage classification was confirmed by TEM observation and sequencing. The host range was determined by the spot test method and double-layer agar plate method. Bacteriostatic ability was evaluated by turbidity measurement. A bacteriophage named L6jm was isolated, which had a regular polyhedron head approximately 75 nm in diameter and a long noncontractile tail with 190 nm in length. As the sequencing result showed no integrase or bacterial genome was in the genome of L6jm, L6jm was defined as lytic bacteriophage. L6jm can infect S. choleraesuis 13312 and Escherichia coli (E. coli) 244. Also, L6jm lysed 9 E. coli strains including one enterotoxigenic E. coli (ETEC) strain. The optimal MOI of L6jm against S. choleraesuis 13122 and E. coli 244 were 0.01 and 0.001 respectively. The latent periods were both 15 min in two strains whereas the rise periods were 145 min and 125 min. L6jm was stable over a wide range of temperatures (30-60 ℃) and pH (3.0-11.0). L6jm could significantly (P<0.05) inhibit the growth of S. choleraesuis 13312, and inhibit ETEC 104 at MOI 100. The polyvalent bacteriophage L6jm infecting S. choleraesuis and E. coli as well as lysing ETEC104 has potential application value in preventing and controlling S. choleraesuis and E. coli infection.

Key words: Salmonella choleraesuis, bacteriophage, polyvalent bacteriophage, characterization of bacteriophage, bacteriostatic effect

CLC Number:

Q939.48

ZHANG Yuyu, ZHANG Min, HUANG Zan, LIN Yan. Isolation and Characterization of a Lytic Polyvalent Bacteriophage Infecting Salmonella choleraesuis and Escherichia coli[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 2032-2038.

References 28

[1]	KONGSTED H, TOFT N, NIELSEN J. Risk factors and epidemiological characteristics of new neonatal porcine diarrhoea syndrome in four Danish herds[J]. BMC Vet Res, 2014, 10(1):151.
[2]	FUNK J A, DAVISE P R, NICHOLS M A. The effect of fecal sample weight on detection of Salmonella enterica in swine feces[J]. J Vet Diagn Invest, 2000, 12(5):412-418.
[3]	ZHANG B Z, KU X G, YU X X, et al. Prevalence and antimicrobial susceptibilities of bacterial pathogens in Chinese pig farms from 2013 to 2017[J]. Sci Rep, 2019, 9(1):9908.
[4]	朱玲云, 郑龙龙, 毕峻龙, 等. 曲靖地区仔猪腹泻细菌性病原的分离鉴定与耐药性分析[J]. 中国畜牧兽医, 2015, 42(3):696-700.ZHU L Y, ZHENG L L, BI J L, et al. Isolation, identification and drug resistance analysis of bacterial pathogen of piglets diarrhea syndrome in Qujing area[J]. China Animal Husbandry & Veterinary Medicine, 2015, 42(3):696-700. (in Chinese)
[5]	LI Y C, PAN Z M, KANG X L, et al. Prevalence, characteristics, and antimicrobial resistance patterns of Salmonella in retail pork in Jiangsu province, Eastern China[J]. J Food Prot, 2014, 77(2):236-245.
[6]	KARUMIDZE N, KUSRADZE I, RIGVAVA S, et al. Isolation and characterisation of lytic bacteriophages of Klebsiella pneumoniae and Klebsiella oxytoca[J]. Curr Microbiol, 2013, 66(3):251-258.
[7]	SAMBROOK J, RUSSELL D W. 分子克隆实验指南[M]. 3版. 黄培堂, 王嘉玺, 朱厚础, 译. 北京:科学出版社, 2002:185-187, 195-197.SAMBROOK J, RUSSELL D W. Molecular cloning:a laboratory manual[M]. 3rd ed. HUANG P T, WANG J X, ZHU H C, trans. Beijing:Science Press, 2002:185-187, 195-197. (in Chinese)
[8]	MIRZAEI M K, NILSSON A S. Isolation of phages for phage therapy:a comparison of spot tests and efficiency of plating analyses for determination of host range and efficacy[J]. PLoS One, 2015, 10(3):e0118557.
[9]	冯烨, 刘军, 孙洋, 等. 噬菌体最新分类与命名[J]. 中国兽医学报, 2013, 33(12):1954-1958.FENG Y, LIU J, SUN Y, et al. An introduction to current classification and nomenclature of bacterial viruses[J]. Chinese Journal of Veterinary Science, 2013, 33(12):1954-1958. (in Chinese)
[10]	WOMMACK K E, COLWELL R R. Virioplankton:Viruses in aquatic ecosystems[J]. Microbiol Mol Biol Rev, 2000, 64(1):69-114.
[11]	DALMASSO M, HILL C, ROSS R P. Exploiting gut bacteriophages for human health[J]. Trends Microbiol, 2014, 22(7):399-405.
[12]	CHAN B K, ABEDON S T, LOC-CARRILLO C. Phage cocktails and the future of phage therapy[J]. Future Microbiol, 2013, 8(6):769-783.
[13]	KUTATELADZE M, ADAMIA R. Bacteriophages as potential new therapeutics to replace or supplement antibiotics[J]. Trends Biotechnol, 2010, 28(12):591-595.
[14]	BARR J J, AURO R, FURLAN M, et al. Bacteriophage adhering to mucus provide a non-host-derived immunity[J]. Proc Natl Acad Sci USA, 2013, 110(26):10771-10776.
[15]	DOORE S M, SCHRAD J R, DEAN W F, et al. Shigella phages isolated during a dysentery outbreak reveal uncommon structures and broad species diversity[J/OL]. J Virol, 2018, 92(8).
[16]	MAHMOUD M, ASKORA A, BARAKAT A B, et al. Isolation and characterization of polyvalent bacteriophages infecting multi drug resistant Salmonella serovars isolated from broilers in Egypt[J]. Int J Food Microbiol, 2018, 266:8-13.
[17]	PARRA B, ROBESON J. Selection of polyvalent bacteriophages infecting Salmonella enterica serovar Choleraesuis[J]. Electron J Biotechnol, 2016, 21:72-76.
[18]	HOWE A, RINGUS D L, WILLIAMS R J, et al. Divergent responses of viral and bacterial communities in the gut microbiome to dietary disturbances in mice[J]. ISME J, 2016, 10(5):1217-1227.
[19]	MANRIQUE P, BOLDUC B, WALK S T, et al. Healthy human gut phageome[J]. Proc Natl Acad Sci USA, 2016, 113(37):10400-10405.
[20]	NORMAN J M, HANDLEY S A, BALDRIDGE M T, et al. Disease-specific alterations in the enteric virome in inflammatory bowel disease[J]. Cell, 2015, 160(3):447-460.
[21]	ABEDON S T. Lysis from without[J]. Bacteriophage, 2011, 1(1):46-49.
[22]	LOESSNER M J. Bacteriophage endolysins——current state of research and applications[J]. Curr Opin Microbiol, 2005, 8(4):480-487.
[23]	HYMAN P, ABEDON S T. Bacteriophage host range and bacterial resistance[J]. Adv Appl Microbiol, 2010, 70:217-248.
[24]	DE JONGE P A, NOBREGA F L, BROUNS S J J, et al. Molecular and evolutionary determinants of bacteriophage host range[J]. Trends Microbiol, 2019, 27(1):51-63.
[25]	CHOI I Y, LEE J H, KIM H J, et al. Isolation and characterization of a novel broad-host-range bacteriophage infecting Salmonella enterica subsp. enterica for biocontrol and rapid detection[J]. J Microbiol Biotechnol, 2017, 27(12):2151-2155.
[26]	CHEN M M, XU J T, YAO H C, et al. Isolation, genome sequencing and functional analysis of two T7-like coliphages of avian pathogenic Escherichia coli[J]. Gene, 2016, 582(1):47-58.
[27]	TANG F, ZHANG P, ZHANG Q M, et al. Isolation and characterization of a broad-spectrum phage of multiple drug resistant Salmonella and its therapeutic utility in mice[J]. Microb Pathog, 2019, 126:193-198.
[28]	AHIWALE S S, BANKAR A V, TAGUNDE S N, et al. Isolation and characterization of a rare waterborne lytic phage of Salmonella enterica serovar Paratyphi B[J]. Can J Microbiol, 2013, 59(5):318-323.

Isolation and Characterization of a Lytic Polyvalent Bacteriophage Infecting Salmonella choleraesuis and Escherichia coli

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