食品科学 ›› 2025, Vol. 46 ›› Issue (6): 89-97.doi: 10.7506/spkx1002-6630-20240925-198

• 生物工程 • 上一篇    下一篇

耐多药奇异变形杆菌噬菌体生物学特性分析及其对鸡胸肉抑菌作用

潘顺圆,陈麒百,王宏月,孔祥宇,庞申雨,高东阳,宋军   

  1. (1.黑龙江八一农垦大学动物科技学院,黑龙江 大庆 163319;2.农业农村部东北寒区牛病防治重点实验室(部省共建),黑龙江 大庆 163319;3.黑龙江省牛病防制重点实验室,黑龙江 大庆 163319;4.大庆市动物源性人兽共患病防控重点实验室,黑龙江 大庆 163319)
  • 出版日期:2025-03-25 发布日期:2025-03-10
  • 基金资助:
    国家自然科学基金青年科学基金项目(31802226);黑龙江省自然科学基金联合引导项目(LH2022C072); 大庆市指导性科技项目(zd-2024-28);黑龙江省牛病防制重点实验室开放课题(PCBD201710)

Biological characteristics of Multidrug-Resistant Proteus mirabilis Bacteriophage and Its Antibacterial Effect on Chicken Breast Meat

PAN Shunyuan, CHEN Qibai, WANG Hongyue, KONG Xiangyu, PANG Shenyu, GAO Dongyang, SONG Jun   

  1. (1. College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China; 2. Key Laboratory of Bovine Disease Control in Northeast China, Ministry of Agriculture and Rural Affairs (Co-construction of Ministry and Province), Daqing 163319, China; 3. Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, Daqing 163319, China; 4. Key Laboratory of Prevention and Control of Zoonotic Diseases of Daqing, Daqing 163319, China)
  • Online:2025-03-25 Published:2025-03-10

摘要: 目的:以耐多药奇异变形杆菌为宿主菌分离噬菌体,测定其生物学特性并评估其对鸡胸肉的抑菌作用。方法:以耐多药奇异变形杆菌为宿主菌分离噬菌体,透射电镜观察噬菌体形态,测定噬菌体感染复数(multiplicity of infection,MOI)、一步生长曲线和噬菌体稳定性;通过点斑法测定噬菌体宿主范围;提取噬菌体基因组并进行全基因组测序分析;采用结晶紫染色法、平板计数法评价噬菌体对生物被膜清除能力;采用平板计数法评价噬菌体对不同材料上生物被膜清除效果,以及测定4 ℃和25 ℃条件下噬菌体对鸡胸肉的抑菌能力。结果:分离获得一株长尾奇异变形杆菌噬菌体vB_PMC-PL1;全基因组分析该噬菌体属于Demerecviridae家族Novosibvirus属,未发现与毒力、抗生素耐药性和整合酶相关的基因;vB_PMC-PL1最佳MOI为0.1、潜伏期为20 min、裂解量为106 PFU/cell;在pH 3~11、4~50 ℃条件下活性稳定;宿主范围显示,vB_PMC-PL1不但能裂解奇异变形杆菌,还能裂解普通变形杆菌;平板计数结果显示vB_PMC-PL1能有效清除不锈钢和高密度聚乙烯上形成的细菌生物被膜。在4 ℃条件下vB_PMC-PL1处理6 h后,鸡胸肉中细菌数量分别减少0.95(lg(CFU/g))(MOI=1 000)和1.01(lg(CFU/g))(MOI=10 000);在25 ℃条件下vB_PMC-PL1处理9 h后细菌数量分别减少1.50(lg(CFU/g))(MOI=1 000)和1.67(lg(CFU/g))(MOI=10 000)。结论:vB_PMC-PL1能够裂解奇异变形杆菌和普通变形杆菌,并对奇异变形杆菌生物被膜具有良好的裂解活性和清除作用,这不仅丰富了变形杆菌噬菌体资源库,而且为变形杆菌噬菌体鸡尾酒的开发与食品生物防控应用提供了参考。

关键词: 奇异变形杆菌;裂解性噬菌体;食源性致病菌;生物被膜;鸡胸肉?

Abstract: Objective: To isolate a bacteriophage infecting multidrug-resistant Proteus mirabilis, determine its biological properties and evaluate its antibacterial effect on chicken breast. Methods: The morphology of the bacteriophage was examined using transmission electron microscopy (TEM), and the multiplicity of infection (MOI), one-step growth curve, and stability were systematically assessed. The host range of the bacteriophage was assessed using the dot blot technique. Subsequently, its genome was extracted and analyzed through whole genome sequencing. Crystal violet staining and plate counting were employed to assess the ability of the phage to remove biofilms and the efficiency in removing biofilms on different materials was evaluated using the plate counting method. The antibacterial effect on chicken breast meat was measured at 4 and 25 ℃. Results: A long-tailed bacteriophage, vB_PMC-PL1, was obtained. Comprehensive genomic analysis revealed that the bacteriophage belonged to the Novosibvirus genus within the Demerecviridae family. Notably, the analysis did not identify any genes associated with virulence, antibiotic resistance, or integrase activity. Phage vB_PMC-PL1 exhibited an optimal MOI of 0.1, an incubation period of 20 min and a burst size of 106 PFU/cell. The titer of vB_PMC-PL1 was stable at pH 3–11 and temperatures ranging from 4 to 50 ℃. The host range test showed that vB_PMC-PL1 could lyse not only Proteus mirabilis but also Proteus vulgaris. vB_PMC-PL1 effectively eradicated bacterial biofilm formed on both stainless steel and high-density polyethylene (HDPE) surfaces. At MOIs of 1 000 and 10 000, it reduced the bacterial concentration in chicken breast meat by 0.95 and 1.01 (lg(CFU/g)) after 6 h incubation at 4 ℃, and by 1.50 and 1.67 (lg(CFU/g)) after 9 h incubation at 25 ℃, respectively. Conclusion: vB_PMC-PL1 exhibited lytic activity against both P. mirabilis and P. vulgaris, demonstrating significant efficacy in disrupting P. mirabilis biofilm. This study not only enriches the Proteus bacteriophage resource library, but also provides a reference for the development of Proteus phage cocktails and the application of biological control in food safety.

Key words: Proteus mirabilis; lytic phage; foodborne pathogen; biofilm; chicken breast meat

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