一株鸡大肠杆菌噬菌体的分离鉴定及治疗试验
|
摘要
为探究大肠杆菌噬菌体的生物学特性及其治疗效果,本研究采用双层平板法分离纯化大肠杆菌噬菌体,并测定噬菌体的效价、温度稳定性、p H稳定性、感染复数(MOI)、一步生长曲线等生物学特性,以及噬菌体治疗大肠杆菌病的效果。结果显示分离的噬菌体Bp1805最佳MOI为1时,其效价可达2.4×10~(10) pfu/m L;噬菌体Bp1805对温度的耐受范围较广,在p H5～p H10范围内非常稳定,该噬菌体潜伏期为20 min,裂解期为50 min。大肠杆菌病治疗试验结果显示,注射噬菌体比口服噬菌体治疗效果好,大肠杆菌病的防治效果与噬菌体治疗方式和治疗时间有关;噬菌体治疗大肠杆菌病效果优于恩诺沙星和硫酸庆大霉素。噬菌体Bp1805能够裂解致病性大肠杆菌,为临床大肠杆菌病的噬菌体防治提供参考依据。
Colibacillosis of chicken is one of the main bacterial infectious diseases harmful to poultry industry and it can bring huge economic losses to poultry industry. To investigate the biological characteristics and therapeutic effect of the bacteriophage on avian colibacillosis, a coliphage was isolated and purified by double-layer plate method, and the characteristics of the isolated coliphage such as titer, temperature stability, pH stability, optimal multiplicity of infection(MOI), one-step growth curve and the therapeutic effect of coliphage on colibacillosis were measured. The results showed that the optimal MOI was 1 and the titer of coliphage Bp1805 was up to 2.4×10~(10) pfu/mL. The temperature tolerance range of Bp1805 is wide, and it was very active in the pH range 5-10. The one step growth curve demonstrated that the incubation period of bacteriophages was 20 min, and lysis period was50 min. The results of the treatment trial of colibacillosis showed that phage injection was better than that of oral medication, and the control effect of colibacillosis is related to the method and time of phage treatment. The effect of Bp1805 on Escherichia coli was better than enoxacin and gentamycin sulfate. The phage Bp1805 could lyse pathogenic Escherichia coli and this study provided theoretical basis for the clinical application of bacteriophage in control of avian colibacillosis.
Colibacillosis of chicken is one of the main bacterial infectious diseases harmful to poultry industry and it can bring huge economic losses to poultry industry. To investigate the biological characteristics and therapeutic effect of the bacteriophage on avian colibacillosis, a coliphage was isolated and purified by double-layer plate method, and the characteristics of the isolated coliphage such as titer, temperature stability, pH stability, optimal multiplicity of infection(MOI), one-step growth curve and the therapeutic effect of coliphage on colibacillosis were measured. The results showed that the optimal MOI was 1 and the titer of coliphage Bp1805 was up to 2.4×10~(10) pfu/mL. The temperature tolerance range of Bp1805 is wide, and it was very active in the pH range 5-10. The one step growth curve demonstrated that the incubation period of bacteriophages was 20 min, and lysis period was50 min. The results of the treatment trial of colibacillosis showed that phage injection was better than that of oral medication, and the control effect of colibacillosis is related to the method and time of phage treatment. The effect of Bp1805 on Escherichia coli was better than enoxacin and gentamycin sulfate. The phage Bp1805 could lyse pathogenic Escherichia coli and this study provided theoretical basis for the clinical application of bacteriophage in control of avian colibacillosis.
引文
[1]高海涛,韩俊丽,关道明.大肠杆菌耐药现状的严峻性[J].生命科学,2017, 29(5):514-520.
[2]周祥莹,张灿,刘文华.鸡白痢沙门氏菌及其噬菌体vB-SpuS-Spp4的分离鉴定[J].病毒学报,2018,34(2):239-243.
[3]吴伟胜.鸡大肠杆菌噬菌体的分离鉴定及初步应用[D].聊城:聊城大学,2016:20-25.
[4]许东勤,姚琳,张奇,等.沙门氏菌和副溶血性弧菌裂解性噬菌体及其裂解酶研究进展[J].食品安全质量检测学报,2017, 8(4):1203-1211.
[5]朱育玮,李玉保,王守荣,等.噬菌体及其治疗细菌感染的研究进展[J].中国畜牧兽医,2015,42(3):769-773.
[6] Zhang Can, Ma Yan-xiang, Wang Ting, et al. Characterization and complete genome sequence of vB EcoP Bp4, a novel polycalent N4 like bacteriophage that infects chicken pathogenic Escherichia coli[J]. Virol Sin, 2016, 31(4):353-356.
[7] Birendra R, Tiwari K, Shukho K, et al. Antibacterial efficacy of lytic Pseudomonas bacteriophage in normal and neutropenic mice models[J]. Microbiology, 2011, 49(6):994-999.
[8] Pouillot F, Chomton M, Blois H, et al. Efficacy of baeteriophage therapy in experimental sepsis and meningitis caused by a clone 025b:H4-ST131 Escherichia coli strain producing CTXM-15[J]. Antimicrob Agents Ch, 2012, 56(7):3568-3575.
[2]周祥莹,张灿,刘文华.鸡白痢沙门氏菌及其噬菌体vB-SpuS-Spp4的分离鉴定[J].病毒学报,2018,34(2):239-243.
[3]吴伟胜.鸡大肠杆菌噬菌体的分离鉴定及初步应用[D].聊城:聊城大学,2016:20-25.
[4]许东勤,姚琳,张奇,等.沙门氏菌和副溶血性弧菌裂解性噬菌体及其裂解酶研究进展[J].食品安全质量检测学报,2017, 8(4):1203-1211.
[5]朱育玮,李玉保,王守荣,等.噬菌体及其治疗细菌感染的研究进展[J].中国畜牧兽医,2015,42(3):769-773.
[6] Zhang Can, Ma Yan-xiang, Wang Ting, et al. Characterization and complete genome sequence of vB EcoP Bp4, a novel polycalent N4 like bacteriophage that infects chicken pathogenic Escherichia coli[J]. Virol Sin, 2016, 31(4):353-356.
[7] Birendra R, Tiwari K, Shukho K, et al. Antibacterial efficacy of lytic Pseudomonas bacteriophage in normal and neutropenic mice models[J]. Microbiology, 2011, 49(6):994-999.
[8] Pouillot F, Chomton M, Blois H, et al. Efficacy of baeteriophage therapy in experimental sepsis and meningitis caused by a clone 025b:H4-ST131 Escherichia coli strain producing CTXM-15[J]. Antimicrob Agents Ch, 2012, 56(7):3568-3575.