探究信都三黄鸡鸡源性噬菌体，明确其生物学特性，为后续噬菌体应用研究做铺垫。本研究从鸡养殖场排泄的污水中分离大肠杆菌，以分离出的大肠杆菌为宿主菌，对养殖场污水中进行噬菌体的初步增殖，采用双层琼脂平板法分离纯化能特异性裂解大肠杆菌的噬菌体,测定其效价后，进一步对其最佳感染复数(MOI)、一步生长曲线、热稳定性和酸碱稳定性等生物学特性进行研究；通过平板菌落计数法测定噬菌体体外抑菌效果。从鸡养殖场污水中分离到21株耐药性大肠杆菌，命名为E1到E2。通过混合平板培养法观察到透明清晰的噬菌斑，分离得到大肠杆菌E13的特异性裂解宿主菌的噬菌体，命名为P1，结果得出P1的最佳感染复数为0.01，一步生长曲线体现为潜伏期40min爆发期50min，温度在40到50℃能保持效价稳定，pH4~11之间存活,且pH为6~7时均保持很高的效价，体外抑菌检测中对宿主菌的生长抑制在前5个小时效果显著，而稳定期的杀菌效果在感染复数为0.001时效果最好。本研究对该噬菌体进行生物学特性及体外抑菌效果评价，为信都三黄鸡致病性大肠杆菌病的防治提供理论依据。

关键词：噬菌体；大肠杆菌；效价；噬菌斑

Abstract:

Investigating avian phages from Xindu Sanhuang chickens to elucidate their biological characteristics, laying the groundwork for subsequent phage application research. This study isolated Escherichia coli from wastewater discharged by chicken farms. Using the isolated E. coli as the host bacterium, phages were preliminarily propagated in the farm wastewater. The double-layer agar plate method was employed to isolate and purify phages capable of specifically lysing E. coli. Following titer determination, further investigations were conducted on biological characteristics including optimal multiplicity of infection (MOI), one-step growth curves, thermal stability, and acid-base stability. In vitro bacteriostatic effects were assessed via plate colony counting. Twenty-one drug-resistant E. coli strains were isolated from poultry farm wastewater, designated E1 to E2. Transparent, well-defined plaques were observed using the mixed-plate culture method. A phage specific to lysing E. coli E13 was isolated and designated P1. Results indicated an optimal multiplicity of infection (MOI) of 0.01 for P1. Its one-step growth curve exhibited a 40-minute latent period and a 50-minute burst phase. The potency remained stable at temperatures between 40 and 50°C, It survived across pH 4–11, maintaining high potency at pH 6–7. In vitro antibacterial assays demonstrated significant growth inhibition of the host bacterium within the first five hours, with optimal lethal effects during the stationary phase at an infection titer of 0.001. This study evaluates the biological characteristics and in vitro antibacterial efficacy of this bacteriophage, providing a theoretical basis for the prevention and control of pathogenic Escherichia coli disease in Xindu Sanhuang chickens.

Keywords: Bacteriophage; Escherichia coli; Potency; Bacteriophage plaque

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参考文献 / References：

何亚丁.噬菌体的生物学特性及其在畜禽健康中的应用[J].当代畜禽养殖业,2020(04):6-8.DOI:10.14070/j.cnki.15-1150.2020.04.002.
何觅之. 大肠杆菌K88、K99广谱噬菌体的分离与生物学特性鉴定[D].华中农业大学,2013.
喻胜猛. 致羊腹泻大肠杆菌噬菌体的分离鉴定及其在环境中的杀菌效果评估[D].西北农林科技大学,2023.DOI:10.27409/d.cnki.gxbnu.2022.000735.
姚澜,张贝贝,王芷洋,等.禽致病性大肠杆菌噬菌体vB_EcoS_AH50的分离鉴定、生物学特性及全基因组分析[J].中国兽医科学,2022,52(12):1568-1577.DOI:10.16656/j.issn.1673-4696.2022.0207.
赵尊福. 鸡源大肠杆菌噬菌体Bp16分离鉴定、生物学特性研究及初步应用[D].西南民族大学,2022.DOI:10.27417/d.cnki.gxnmc.2021.000045.
郝贺,张永英,钟翠红,等.鸡源沙门菌噬菌体的生物学特性及其对雏鸡肠道结构的影响[J].中国兽医科学,2022,52(02):259-268.DOI:10.16656/j.issn.1673-4696.2022.0025.
艾梦燕. 鸡源致病性大肠杆菌噬菌体的分离鉴定及生物学特性研究[D].河南农业大学,2023.DOI:10.27117/d.cnki.ghenu.2022.000335.
徐美余,张瑶,邓征宇,等.一株可裂解动物源性多重耐药志贺氏菌的噬菌体分离鉴定及其生物学特性研究[J].微生物学杂志,2023,43(05):91-100.
吴圆圆,屈勇刚,于会举,等.一株鸡大肠埃希氏菌裂解性噬菌体的分离与生物学特性分析[J].动物医学进展,2021,42(05):30-35.DOI:10.16437/j.cnki.1007-5038.2021.05.007.
王嘉威,贺永超,乔元明,等.噬菌体特性及应用研究进展[J].山东畜牧兽医,2024,45(03):77-80
Huff W E, Huff G R, Rath N C, et al. Bacteriophage Treatment of a Severe Escherichia coli Respiratory Infection in Broiler Chickens[J]. Avian Diseases, 2003,47(4): 1399-1405.
Oliveira A, Sereno R, Azeredo J. In vivo efficiency evaluation of a phage cocktail in controlling severe colibacillosis in confined conditions and experimental poultry houses[J]. Veterinary Microbiology, 2010, 146(3):303-308.
高铎,王晋宇,李河谕,等.1株肠炎沙门菌噬菌体GD01的分离鉴定及生物学特性分析[J].中国畜牧兽医,2023,50(12):5160-5169.DOI:10.16431/j.cnki.1671-7236.2023.12.037.