微生物学报

Phage	Antibiotic	Pathogenic bacteria	Results
T1245	Ceftazidime, colistin, imipenem, and meropenem	Acinetobacter baumannii	PAC significantly reduced bacterial density by up to 80.0%, disrupted biofilm structure, and altered cell morphology. Additionally, it notably decreased the biomass and bacterial viability of 3-day-old biofilms^[21]
PH826	Ceftazidime and colistin	Pseudomonas aeruginosa	PAC reduced biofilm formation by more than 80.0%^[22]
Remus	Vancomycin	Staphylococcus aureus	In comparison to the use of Remus or vancomycin alone, PAC exhibited greater efficacy in reducing bacterial load. Furthermore, when tested in vivo, PAC achieved the highest survival rate^[23]
Klebsiella-specific phage	Imipenem	Klebsiella pneumoniae	PAC demonstrates a marginal improvement in efficacy compared to phage treatment alone, potentially inhibiting the regrowth of bacteria resistant to the phage to a certain extent^[24]
StM171	Ampicillin, chloramphenicol, levofloxacin, tetracycline, and gentamicin	Stenotrophomonas maltophilia	In most instances, StM171 augmented the inhibitory effect exerted by antibiotics^[25]
vB_SauM-A and vB_SauM-D	Ciprofloxacin	Staphylococcus aureus and Candida albicans	PAC achieved a substantial reduction of 90.0% in biofilm-specific activity for mono-species biofilms and 69.0% for dual-species biofilms, outperforming single phage or antibiotics alone^[26]
vB_Eco4M-7 and ECML-117	Rifampicin and ciprofloxacin	Escherichia coli	PAC effectively reduces the bacterial count and biofilm density. Furthermore, under PAC treatment, the efficiency of emergence of bacteria resistant to phages and rifampicin was significantly diminished^[27]

Phage	Antibiotic	Pathogenic bacteria	Results
T1245	Ceftazidime, colistin, imipenem, and meropenem	Acinetobacter baumannii	PAC significantly reduced bacterial density by up to 80.0%, disrupted biofilm structure, and altered cell morphology. Additionally, it notably decreased the biomass and bacterial viability of 3-day-old biofilms^[21]
PH826	Ceftazidime and colistin	Pseudomonas aeruginosa	PAC reduced biofilm formation by more than 80.0%^[22]
Remus	Vancomycin	Staphylococcus aureus	In comparison to the use of Remus or vancomycin alone, PAC exhibited greater efficacy in reducing bacterial load. Furthermore, when tested in vivo, PAC achieved the highest survival rate^[23]
Klebsiella-specific phage	Imipenem	Klebsiella pneumoniae	PAC demonstrates a marginal improvement in efficacy compared to phage treatment alone, potentially inhibiting the regrowth of bacteria resistant to the phage to a certain extent^[24]
StM171	Ampicillin, chloramphenicol, levofloxacin, tetracycline, and gentamicin	Stenotrophomonas maltophilia	In most instances, StM171 augmented the inhibitory effect exerted by antibiotics^[25]
vB_SauM-A and vB_SauM-D	Ciprofloxacin	Staphylococcus aureus and Candida albicans	PAC achieved a substantial reduction of 90.0% in biofilm-specific activity for mono-species biofilms and 69.0% for dual-species biofilms, outperforming single phage or antibiotics alone^[26]
vB_Eco4M-7 and ECML-117	Rifampicin and ciprofloxacin	Escherichia coli	PAC effectively reduces the bacterial count and biofilm density. Furthermore, under PAC treatment, the efficiency of emergence of bacteria resistant to phages and rifampicin was significantly diminished^[27]

Model	Phage or endolysin	Pathogenic bacteria	Antibiotic	Result
Zebrafish infection model	YC#06	Acinetobacter baumannii	Antibiotic mixtures (chloramphenicol, imipenem, and cefotaxime)	PAC significantly increased the survival rate of zebrafish^[33]
Galleria mellonella larvae survival model	KP1801	Klebsiella pneumoniae	None	Bacteriophage significantly improved the survival rate of larvae. The number of intracellular bacteria in larvae showed a significant decrease while the number of phage increased^[34]
Zebrafish infection model	KpG	Klebsiella pneumoniae	Streptomycin	Treatment of zebrafish muscle tissue with KpG phage and PAC showed a significant 77.7% and 97.2% decline in CFU/ml, respectively^[35]
Galleria mellonella invertebrate infection model	ɸWL-3	Escherichia coli	Fosfomycin	PAC can improve the survival rate of larvae^[36]
Caenorhabditis elegans	Motto	Pseudomonas aeruginosa	None	A 90% survival rate when treated with the phage at a multiplicity of infection of 10.0^[12]
Mice infection model	vB_PaeP_PS28	Pseudomonas aeruginosa	None	After phage treatment, the mortality rate decreased significantly and the bacterial colonization in organs was reduced^[37]
Galleria mellonella model	vB_SauM-D	Staphylococcus aureus	None	Within 60 hours, the control group exhibited 100% lethality, whereas the larvae treated with phage displayed a survival rate of 86% after 120 hours^[38]
Spn mouse nasopharyngeal colonization model	SP-CHAP (endolysin)	Streptococcus pneumoniae	None	The application of SP-CHAP resulted in a notable decrease in the colony-forming units of Spn, surpassing even the effectiveness of Cpl-1^[39]
A mouse model of Streptococcus uberis mastitis	NC5 (VersaTile engineered endolysin)	Streptococcus uberis	Cloxacillin	The high-dose combination therapy significantly reduced the hallmarks of mastitis in fast responders, exhibiting a 13 000-fold reduction in bacterial load, a 5.7-fold decrease in neutrophil infiltration, and a 13.0-fold decrement in the key pro-inflammatory chemokine IL-8^[40]

Model	Phage or endolysin	Pathogenic bacteria	Antibiotic	Result
Zebrafish infection model	YC#06	Acinetobacter baumannii	Antibiotic mixtures (chloramphenicol, imipenem, and cefotaxime)	PAC significantly increased the survival rate of zebrafish^[33]
Galleria mellonella larvae survival model	KP1801	Klebsiella pneumoniae	None	Bacteriophage significantly improved the survival rate of larvae. The number of intracellular bacteria in larvae showed a significant decrease while the number of phage increased^[34]
Zebrafish infection model	KpG	Klebsiella pneumoniae	Streptomycin	Treatment of zebrafish muscle tissue with KpG phage and PAC showed a significant 77.7% and 97.2% decline in CFU/ml, respectively^[35]
Galleria mellonella invertebrate infection model	ɸWL-3	Escherichia coli	Fosfomycin	PAC can improve the survival rate of larvae^[36]
Caenorhabditis elegans	Motto	Pseudomonas aeruginosa	None	A 90% survival rate when treated with the phage at a multiplicity of infection of 10.0^[12]
Mice infection model	vB_PaeP_PS28	Pseudomonas aeruginosa	None	After phage treatment, the mortality rate decreased significantly and the bacterial colonization in organs was reduced^[37]
Galleria mellonella model	vB_SauM-D	Staphylococcus aureus	None	Within 60 hours, the control group exhibited 100% lethality, whereas the larvae treated with phage displayed a survival rate of 86% after 120 hours^[38]
Spn mouse nasopharyngeal colonization model	SP-CHAP (endolysin)	Streptococcus pneumoniae	None	The application of SP-CHAP resulted in a notable decrease in the colony-forming units of Spn, surpassing even the effectiveness of Cpl-1^[39]
A mouse model of Streptococcus uberis mastitis	NC5 (VersaTile engineered endolysin)	Streptococcus uberis	Cloxacillin	The high-dose combination therapy significantly reduced the hallmarks of mastitis in fast responders, exhibiting a 13 000-fold reduction in bacterial load, a 5.7-fold decrease in neutrophil infiltration, and a 13.0-fold decrement in the key pro-inflammatory chemokine IL-8^[40]

噬菌体防治细菌生物被膜相关感染的研究进展

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贺丽莎 ¹ , 杨灼 ¹^,² , 冉蕾 ¹^,² , 陈红伟 ¹^,²^,³^,⁴^,^* , 杨洪早 ¹^,²^,³^,⁴^,^*

微生物学报 | 综述 2024,64(10): 3670-3684

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微生物学报 | 综述 2024, 64(10): 3670-3684

噬菌体防治细菌生物被膜相关感染的研究进展

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贺丽莎¹, 杨灼¹^,², 冉蕾¹^,², 陈红伟¹^,²^,³^,⁴^,^*, 杨洪早¹^,²^,³^,⁴^,^*

作者信息

1 西南大学动物医学院, 重庆 402460

2 国家生猪技术创新中心, 重庆 402460

3 西南大学医学研究院, 免疫学研究中心, 重庆 402460

4 西南大学, 中兽医药研究所, 重庆 402460

Research progress of phages in prevention and treatment of bacterial biofilm-associated infections

Lisha HE¹, Zhuo YANG¹^,², Lei RAN¹^,², Hongwei CHEN¹^,²^,³^,⁴^,^*, Hongzao YANG¹^,²^,³^,⁴^,^*

Affiliations

1 College of Veterinary Medicine, Southwest University, Chongqing 402460, China

2 National Center of Technology Innovation for Pigs, Chongqing 402460, China

3 Immunology Research Center, Medical Research Institute, Southwest University, Chongqing 402460, China

4 Traditional Chinese Veterinary Research Institute, Southwest University, Chongqing 402460, China

出版时间: 2024-08-06 doi: 10.13343/j.cnki.wsxb.20240259

文章导航

摘要

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细菌生物被膜是由细菌产生的胞外多聚物围绕而成的聚集体，其对抗菌剂和宿主免疫防御机制展现出显著抗性，成为临床细菌感染疾病难以根治的关键因素。噬菌体作为一类能够特异性感染并裂解细菌的病毒，在防治生物被膜相关感染领域展现出巨大潜力。本文综述了噬菌体、噬菌体内溶素以及噬菌体协同抗生素防治生物被膜相关感染的作用效果、体内外研究方法与作用机理，并探讨了噬菌体在防治生物被膜相关感染中的前景与潜在障碍，同时进行展望，旨在为开发更高效的治疗药物提供有价值的参考。

关键词

噬菌体 / 细菌生物被膜 / 内溶素 / 噬菌体-抗生素联合疗法

Abstract

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Bacterial biofilms are aggregates surrounded by extracellular polymeric substances produced by bacteria, exhibiting significant resistance to antimicrobials and host immune defense mechanisms. As a result, they become a crucial factor in the recalcitrance of bacterial infections. Phages, as a class of viruses capable of specifically infecting and lysing bacteria, have demonstrated immense potential in the prevention and treatment of biofilm-associated infections. This review summarizes the efficacy, in vitro and in vivo research methods, and mechanisms of phages, phage endolysins, and phage-antibiotic combinations in combating biofilm-associated infections. Furthermore, it discusses the prospects and potential obstacles of phages in this field, aiming to give insights into the development of more effective therapeutic agents.

Key words

phage / bacterial biofilm / endolysin / phage-antibiotic combination therapy

引用本文

贺丽莎, 杨灼, 冉蕾, 陈红伟, 杨洪早. 噬菌体防治细菌生物被膜相关感染的研究进展. 微生物学报, 2024 , 64 (10) : 3670 -3684 . DOI: 10.13343/j.cnki.wsxb.20240259

Lisha HE, Zhuo YANG, Lei RAN, Hongwei CHEN, Hongzao YANG. Research progress of phages in prevention and treatment of bacterial biofilm-associated infections[J]. Acta Microbiologica Sinica, 2024 , 64 (10) : 3670 -3684 . DOI: 10.13343/j.cnki.wsxb.20240259

正文

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细菌生物被膜是指附着在生物体或者非生物体表面并牢固地嵌入胞外聚合物(extracellular polymeric substance, EPS)基质中的细菌的复杂三维聚集，其形成通常分为附着、定植、发育、成熟以及分散5个阶段，其中分散是新一轮生物被膜形成的开始^[1]。

生物被膜在环境中普遍存在，相较于浮游细菌，其内的细菌对抗菌剂和免疫机制的敏感性显著降低，因此展现出更强的致病性和抵抗力^[2]。临床上，生物被膜常导致持续性感染^[3]；在工业领域，它则加剧了腐蚀过程并增加了有害副产物的释放^[4]。此外，生物被膜为食源性病原体提供了“保护屏障”，从而减少病原菌与抗菌物质的接触，对食品安全构成严重威胁^[4]。尤为值得关注的是，生物被膜的形成使得细菌对抗生素的耐药性提高约1 000倍，这无疑增加了生物被膜清除的难度^[5]。

在积极寻找有效抗生物被膜感染策略的过程中，噬菌体、螯合剂、超声波、群体感应(quorum sensing, QS)抑制剂和植物源性活性物质等均显示出优良抗生物被膜效果，如本课题组Zhang等^[6-7]改良设计的鼠源抗微生物肽具有显著清除铜绿假单胞菌生物被膜的能力。其中，噬菌体与其他物质相比，具有专一性强、残留少、自我增殖快、用量少且具有潜在可修饰性等优点，是防治生物被膜相关感染的有力武器^[8-9]。本文结合当前时代背景与专业研究进展，概述了噬菌体、噬菌体内溶素以及噬菌体协同抗生素防治生物被膜相关感染的作用效果、体内外研究方法与作用机理，旨在为完善抗生物被膜策略、寻找高效治疗药物提供参考。

1 作用效果

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1.1 噬菌体及内溶素作用效果

噬菌体作为特异性感染细菌的病毒，根据其感染宿主细菌的方式，可分为温和噬菌体和烈性噬菌体^[10]。研究表明，噬菌体T4D⁺能够成功感染大肠杆菌3000 XIII生物被膜内细菌，这是首次观察到噬菌体对生物被膜具有溶解性感染能力^[11]。随后，众多研究相继证实，噬菌体具有显著抑制多种病原菌生物被膜形成的作用。以铜绿假单胞菌为例，Manohar等^[12]研究发现，噬菌体Motto减少了至少75.0%的生物被膜，具有显著抗生物被膜活性。值得注意的是，噬菌体混合物的应用展现出与单一噬菌体同等甚至更佳的抗生物被膜感染效果，进一步突显了噬菌体的巨大潜力^[13]。

此外，噬菌体在裂解后期依赖宿主细菌合成并分泌内溶素，这是一种由噬菌体编码的水解酶，对多种病原菌具有致死作用，目前尚未有报道显示其对人类具有直接毒性^[14]。近年来，对噬菌体内溶素的研究主要聚焦于金黄色葡萄球菌、肺炎克雷伯氏菌及铜绿假单胞菌等。如Soontarach等^[15]研究发现，噬菌体LysAB1245能显著减少铜绿假单胞菌和金黄色葡萄球菌形成的生物被膜，并且呈浓度依赖性。另一方面，噬菌体内溶素具有比噬菌体更广泛的抗菌谱，细菌难以对内溶素产生耐药性^[16]。同时，在一定限度下，随着内溶素使用浓度和时间的增加，抑制效果往往增强^[17]。此外，噬菌体内溶素的应用已经从单一物种生物被膜的对抗扩展到多物种生物被膜的防控，如内溶素Ply113已被证实能抑制由粪肠球菌与金黄色葡萄球菌共同形成的双物种生物被膜^[18]。

1.2 噬菌体协同抗生素作用效果

噬菌体-抗生素联合疗法(phage-antibiotic combination therapy, PAC)是治疗生物被膜疾病潜在的抗生素替代方法。在抗生素的亚抑菌浓度下，噬菌体-抗生素协同作用(phage-antibiotic synergy, PAS)使细菌中后代噬菌体释放增多，并能降低抗生素对生物被膜的最小根除浓度，减少抗生素的使用量^[19]。然而，为达到最好的联合应用效果，应用PAC时必须考虑两者的给药顺序，不同组合、不同治疗目的的给药顺序不同，序贯治疗往往显示出高效的抗生物被膜感染能力^[20]。相较于单独使用抗生素或噬菌体，PAC在破坏生物被膜方面展现出了更为出色的能力(表1)。特别是在多物种生物被膜中，PAC显著减少细菌负荷的特性表现得尤为明显^[26]。此外，PAC还具有其他优点，如有效延长噬菌体的存活时间、显著减缓抗生素耐药性的发展以及降低耐噬菌体细菌的产生等^[20]。

2 研究方法

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2.1 体外研究

噬菌体防控生物被膜相关感染的体外研究主要包括2个方面：(1) 在具备生物被膜形成条件的环境中进行试验；(2) 将原位生长的生物被膜转移到体外环境中进行进一步的研究。

在具备生物被膜形成条件的环境中进行试验，避免了体内复杂因素的干扰，使结果更为客观且操作更为简便。例如，在培养基或96孔微量滴定板中构建生物被膜，利用染料(如结晶紫、二甲基亚甲基蓝)或指示细菌活力的染色剂(如刃天青、可溶性四唑鎓)进行染色，随后通过微孔板分光光度计测量特定波长处的光密度值，从而量化生物被膜的生物量，并据此评估生物被膜的形成水平(非生物被膜、弱生物被膜、中等生物被膜和强生物被膜)、生物被膜形成百分比与细菌活力等^[15]。此外，激光扫描共聚焦显微镜技术结合Biofilm Q软件，能够直观展现生物被膜的结构和厚度，为评估噬菌体的作用效果提供有力支持^[28-29]。同时，通过菌落形成单位(colony-forming units, CFU)计数能测定细菌存活率，从而直观反映噬菌体对生物被膜的影响^[23]。

另一方面，将原位生长的生物被膜转移到体外环境中进行进一步研究，如人尿模型和猪皮肤外植体模型等，为噬菌体的抗生物被膜感染作用提供了更为接近真实生理环境的模拟条件。例如，在人尿模型中，PAC显著减少了鲍曼不动杆菌生物被膜的生物量^[30]；在猪皮肤外植体模型中，噬菌体pB3074与头孢噻肟或美罗培南的联合使用，对离体伤口感染的治疗表现出显著效果^[31]。这种方法不仅使体外研究更加贴近体内环境，而且便于比较原位与体外生长的生物被膜，从而更准确地评估实验的真实性和有效性。

由于体外实验条件可控且周期短，能够快速准确地评估噬菌体对生物被膜的作用效果，为体内研究提供了有力的方法和数据参考。然而，当前体外研究仍存在一些局限性，如新生物被膜生长与现有生物被膜去除的区分不明确、未有效排除EPS解聚酶等其他物质对生物被膜的影响、浮游细菌能增加噬菌体滴度从而影响实验结果以及模拟体内真实情况存在误差等^[32]。未来研究可考虑在噬菌体处理前(即零时间)测量生物被膜特性、提高噬菌体滴度或多次给药、使用流动条件去除浮游细菌等方法来减少或消除这些不足。

2.2 体内研究

为提高研究数据的准确性和临床实验的实用性，体内研究显得尤为重要。在这一环节中，研究者通常会先构建动物模型(表2)，以便在体内模拟生物被膜的形成过程。随后，给药处理并结合临床症状，采集样本菌进行生物被膜染色(如结晶紫)和菌落形成单位计数，从而直观地反映噬菌体对生物被膜的体内治疗效果。同时，利用小动物活体成像仪等工具观察动物的存活率和组织变化，间接体现噬菌体的作用效果。例如，Fursov等^[17]采用植入式扩散室方法构建大鼠肺炎克雷伯氏菌生物被膜模型，通过测量生物被膜活菌计数值与观察腔室周围组织的炎症反应判断内溶素LysECD7的体内效果。另外，Łusiak-Szelachowska等^[41]通过创建绵羊鼻窦炎模型，利用光密度值与生物被膜厚度的观察，探究了噬菌体对金黄色葡萄球菌和铜绿假单胞菌生物被膜的体内抑制效果。

诸多研究表明，噬菌体能显著降低动物器官组织中细菌负荷，提高试验动物存活率，并且减少组织损伤。然而，这种疗效会随着细菌攻击后再使用噬菌体的时间间隔延长而降低^[42]。值得注意的是，PAC在体内应用中往往展现出更好的治疗效果，并且具有特异性^[36]；噬菌体内溶素在体内作用效果显著，通过基因工程改造，可以进一步增强其裂解活性并扩大宿主谱^[14]。

体内研究能够真实、准确地反映出噬菌体在体内对抗生物被膜感染的情况，为临床研究提供了坚实可靠的依据。然而，与体外研究相比，体内研究往往成本高昂、耗时长且操作难度较大，此外，其所涉及的病原菌种类相对单一，限制了其应用范围。因此，未来的研究可尝试结合基因工程技术进一步探索不同噬菌体在体内对抗多种病原菌的效果，并更新和丰富动物模型种类。

3 作用机理

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3.1 噬菌体防治细菌生物被膜相关感染的作用机理

噬菌体作为地球上一种极为丰富的微生物资源，其独特的破坏细菌生物被膜的作用机理显著区别于传统的抗生素和消毒剂。噬菌体不仅能高效抗细菌生物被膜，而且能有效作用于生物被膜内细菌，并展现出灵活多变的应对策略，为细菌生物被膜相关感染的防控提供新的思路和策略(图1)。

3.1.1 酶降解

噬菌体的基因组能编码合成多种酶，如内溶素、解聚酶以及与病毒粒子相关的肽聚糖水解酶等，这些酶能降解细菌肽聚糖、荚膜多糖等物质，协助噬菌体破坏细菌生物被膜的完整性从而促进噬菌体高效穿透生物被膜^[43-44]。在这些酶中，内溶素和解聚酶尤其重要，构成了噬菌体防治生物被膜相关感染的主要基础^[45]。

内溶素作为噬菌体复制周期的最后产物，能够特异性地降解细菌细胞壁中的肽聚糖，从而有效抑制生物被膜的形成，使噬菌体快速渗透生物被膜^[43]。这种酶通常编码在噬菌体的内部或尾部区域，使得它能够从细菌的内部和外部2个方向切割肽聚糖^[46]。

解聚酶的结构域则常以尾丝的形式展示在噬菌体的顶端，以尾刺蛋白或游离酶的形式降解荚膜多糖、脂多糖、O型多糖链或生物被膜外多糖，进而削弱生物被膜结构，促进噬菌体扩散^[47]。

值得一提的是，噬菌体不仅自身能够编码解聚酶，还能诱导宿主细菌产生EPS解聚酶。这种酶能够高效地分解EPS中的多糖和蛋白质，通过其裂解活性，进一步促进噬菌体渗透、复制^[48]。

3.1.2 利用生物被膜水通道扩散

在生物被膜的结构中，存在一类特殊的水通道，这些通道因尺寸限制而阻碍了体积较大的抗菌物质的渗透；然而，噬菌体却能凭借其较小的体积直接通过这些水通道进行扩散^[49]。在扩散过程中，噬菌体不仅利用重力作用穿透生物被膜的内部层次，而且其数量不断增加，无需消耗自身浓度^[50]。因此，相较于抗生素，噬菌体在扩散时更为高效。

3.1.3 利用载体细菌穿透生物被膜

在微生物群落生态系统中，噬菌体展现了一种独特的吸附机制，能够非特异性或特异性地附着于具有活动性的细菌(载体)表面，从而渗透到生物被膜内部^[48]。值得注意的是，载体细菌的运动机制促进了噬菌体向生物被膜内营养富集区域的定向迁移，相比于随机的扩散行为，这种迁移具有定向性与高效性^[49]。

3.1.4 影响细菌群体感应系统

群体感应作为细菌群体行为调控的关键机制，涉及细菌基因表达的调控以及生物被膜内部特定小分子信号(即自诱导物)的生成和检测^[51]。当生物被膜受到外界压力(如噬菌体)时，QS系统可通过特定的信号分子精细调控多糖、胞外DNA等关键物质的合成基因表达，进而维护生物被膜的稳定性^[52]。然而，当细菌通过调控QS系统抵抗噬菌体的感染时，噬菌体也能产生QS抑制剂破坏这一信息传导途径。以铜绿假单胞菌噬菌体DMS3为例，其编码的新型QS抗激活蛋白Aqs1能抑制群体感应的主要调节因子LasR，影响QS系统介导的多种抗噬菌体策略；同时，Aqs1还能与Ⅳ型菌毛组装蛋白PilB相互作用并抑制PilB，影响细菌抽搐运动，降低细菌毒力^[53]。此外，噬菌体除了能释放QS抑制剂外，还能借助QS信号分子进入裂解程序。如霍乱弧菌噬菌体VP882能编码弧菌QS受体转录因子的同源物，即受体VqmA，VqmA与弧菌QS系统信号分子3, 5-二甲基吡嗪-2-醇结合后不仅能激活宿主基因VqmR的转录从而影响弧菌QS途径，还能激活噬菌体表达抗阻遏蛋白Qtip从而启动噬菌体裂解程序^[54]。因此，噬菌体可通过多种手段调控生物被膜内细菌QS信号分子甚至借助QS信号分子诱导噬菌体进入裂解程序，从而阻断QS级联反应，抑制生物被膜的形成与成熟。

值得关注的是，QS系统在生物被膜分散中发挥着重要作用。以铜绿假单胞菌的QS系统Las I/Las R为例，该系统通过正向调节酪氨酸磷酸酶A的合成来抑制基因pel的表达并降低环鸟苷二磷酸(cyclic dimeric guanosine monophosphate, c-di-GMP)水平，从而促进生物被膜分散，其中降低胞内c-di-GMP水平已被认为是根除生物被膜的新策略^[1]。同时，本课题组Zhang等^[6]研究发现经过改良设计的鼠源抗微生物肽(cathelicidin related antimicrobial peptide, CRAMP)也具有显著降低c-di-GMP水平的能力，此外，CRAMP还能够显著上调参与铜绿假单胞菌喹诺酮信号系统合成的相关基因或蛋白的表达水平，从而增加铜绿假单胞菌(实验室菌株PAO1)生物被膜鼠李糖脂含量，其中鼠李糖脂的产生增加有助于铜绿假单胞菌PAO1生物被膜的扩散，因此CRAMP展现出优异的生物被膜分散能力。那么，噬菌体是否能通过调控QS系统来影响生物被膜中鼠李糖脂乃至c-di-GMP的含量，进而成为一种潜在的生物被膜分散剂？展望未来，相信相关研究将进一步深入揭示这一领域的更多可能性。

3.1.5 拦截持留菌

持留菌是指存在于生物被膜中或浮游细菌中的短暂的抗生素耐受和休眠亚群^[49]。抗生素主要针对代谢活跃的细菌产生杀菌作用，但治疗中断后，进入到休眠状态的持留菌可能被重新激活，导致生物被膜反复形成^[55]。与之相比，噬菌体展现出显著优势，不仅能有效感染并杀灭处于休眠状态的持留菌，而且在侵入宿主细菌后，能进入一种类似细菌休眠的可逆非活跃状态，等待营养恢复后重新激活，继续其裂解细菌和瓦解生物被膜的功能^[49]。

3.1.6 克服细菌RM和CRISPR-Cas防御系统

限制性修饰系统(restriction modification system, RM)和成簇规律间隔短回文重复序列及相关蛋白(clustered regularly interspaced short palindromic repeats and CRISPR-associated protein, CRISPR-Cas)系统是细菌广为人知的2种防御机制，这2种系统共同在特定基因位点识别和切割噬菌体DNA，以保护细菌自身的基因组免受侵害^[56]。然而，噬菌体也通过进化发展出了一系列逃逸细菌免疫系统的策略。

RM系统通常依赖限制性核酸内切酶(restriction enzyme, REase)在特定的识别位点切割外源DNA^[57]。面对这一威胁，噬菌体展现出多种逃避策略，如通过甲基转移酶(methyltransferase, MTase)修饰其基因组以伪装成宿主DNA、与基因组共同注射特定的蛋白质(如噬菌体P1编码的DarA和DarB)以遮蔽限制性位点，以及编码特定的蛋白质(如噬菌体λ编码的抗限制蛋白Ral)来增强MTase活性并抑制REase活性等^[58]。此外，一些噬菌体通过在其基因组中拥有较少的限制性位点，或者这些位点之间距离过远而无法被宿主REase有效识别，从而巧妙地避免了成为细菌RM系统的攻击目标^[57]。

另一种防御机制CRISPR-Cas系统由CRISPR位点和Cas基因组成，是截至目前已知的唯一适应性和可遗传的防御系统^[59]。然而，噬菌体已经进化出多种策略来对抗这一防御系统。首先，噬菌体可以编码抗CRISPR蛋白(如Acr蛋白)，这些蛋白能够直接使CRISPR-Cas免疫蛋白失活，从而削弱细菌的免疫能力^[60]。此外，噬菌体还能通过原间隔物相邻基序或种子序列中的点突变、原间隔区的缺失以及原间隔区DNA的修饰等手段，有效避免CRISPR RNA的识别，进而绕过细菌的CRISPR-Cas防御^[49]。更为独特的是，巨型噬菌体甚至能产生核样结构，这一结构能够遏制Cas复合物的形成，阻止细菌对噬菌体DNA的靶向识别和切割^[61]。

研究表明噬菌体基因组的共价修饰(如羟甲基化和葡萄糖基化)也赋予了细菌CRISPR系统一定程度的抗性，并允许突变噬菌体在CRISPR压力下快速进化^[56]。这一发现揭示了细菌RM和CRISPR-Cas防御系统之间存在紧密的相互关联，而非孤立存在。更重要的是，这一现象为噬菌体逃避细菌免疫系统，进而对抗生物被膜感染提供了一种潜在的机制。

3.2 噬菌体协同抗生素防治细菌生物被膜相关感染的作用机理

当噬菌体与抗生素联合使用时，研究人员提出了多种作用机制以阐释其协同作用：(1) EPS具有阻碍抗生素扩散的效应，然而噬菌体能够穿透EPS并降解细菌细胞壁，从而促进抗生素的渗透和效用发挥^[62]；(2) 适量抗生素能增加噬菌斑大小与噬菌体裂解量，如哌拉西林和头孢他啶已被证实能显著提升铜绿假单胞菌噬菌体KPP22的噬菌斑大小，使其增加至原先的2−3倍^[63]；(3) 抗生素的作用会促使细菌发生丝状化，这种变化进而扰动肽聚糖层，使得细菌对噬菌体编码的裂解酶的作用变得更为敏感^[64]；(4) 抗生素与噬菌体联合使用有效减缓了抗生素耐药性的发展，并显著减少噬菌体抗性突变体的产生^[20]；(5) 噬菌体和抗生素的协同作用也可能是由于抗生素最低抑菌浓度的降低^[65]。

然而，关于噬菌体与抗生素在生物被膜内相互作用并影响宿主免疫应答的具体机制，目前尚缺乏明确的研究成果，有待进一步深入研究。

4 应用前景及潜在障碍

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4.1 应用前景

在医疗领域，噬菌体能显著抑制并破坏组织器官上的细菌生物被膜。例如，噬菌体及其酶能显著减少由口腔细菌如放线菌属、梭杆菌属及金黄色葡萄球菌等形成的生物被膜，从而有效防治龋齿、牙周病等口腔疾病^[66]。此外，噬菌体有助于清除植入材料上的生物被膜，如噬菌体Isf-Pm1与噬菌体Isf-Pm2的混合制剂能显著控制奇异变形杆菌在导管相关尿路感染中的表面定植^[67]。在其他领域，噬菌体也展现出显著的价值。如噬菌体AZO145A能有效减少不锈钢上产志贺毒素大肠杆菌O145生物被膜的形成，进而降低牛肉的污染风险^[68]；噬菌体T7、SPI1及GTE7能够有效去除废水中的生物被膜，从而显著提升废水处理的效率^[69]。

目前，噬菌体治疗的高效性已得到实际应用，而研究者们进一步改良设计的工程化噬菌体疗法更是展现出巨大应用潜力。以丝状大肠杆菌噬菌体M13为例，经过多肽修饰的M13噬菌体相较于单独使用野生多价噬菌体，在减少活细菌生物量和降低生物被膜表面覆盖率方面展现出更为显著的效果，并且由于噬菌体改造技术允许根据特定使用场景进行特异性修饰，因此可以实现靶向性的灵活调整^[70]。此外，噬菌体工程方法种类丰富，如同源重组、电穿孔介导的噬菌体重组、CRISPR基因编辑技术、体内重组以及基于酵母的噬菌体基因组组装工程等，这些方法在高级基因组层面上对噬菌体进行精确修改，从而赋予噬菌体各种独特的能力^[71]。除了工程化噬菌体外，研究者还开发了多种包封递送技术^[72]。例如，Wroe等^[73]成功设计了一种可注射的水凝胶，该水凝胶能够有效封装铜绿假单胞菌噬菌体，并能调节噬菌体递送至骨感染部位的释放速率，封装在水凝胶中的噬菌体与游离噬菌体相比具有更高的抗生物被膜效率。由此可见，噬菌体疗法正逐渐融合多种先进技术，在实际应用中，结合包封递送技术与噬菌体工程化，或许能为产业型研究人员提供新的实践方向。然而，多种技术的结合是否能达到“强强联合”的效果？这值得在未来的研究中深入探讨。

4.2 潜在障碍

噬菌体疗法在防治细菌生物被膜相关感染领域取得了显著成效，然而，其临床应用仍面临一些挑战和局限性。首先，噬菌体的抗菌谱相对较窄，这一特性使其能够精准地抑制生物被膜的形成，同时避免对正常菌群的干扰，但在治疗涉及多种生物被膜的感染时，还需进行噬菌体的选择，这可能会在一定程度上影响临床治疗的效率和速度^[74]。其次，细菌在面对噬菌体攻击时会产生抵抗性，尽管研究表明PAS能有效减少耐药细菌的出现，但细菌的噬菌体抗性问题仍尚未完全解决^[75]。此外，噬菌体进入机体后，会被机体的免疫系统识别并清除，这一过程中可能涉及体液免疫和细胞免疫的激活^[76]。因此，如何平衡噬菌体的治疗效果与机体的免疫反应，是噬菌体疗法在临床应用中需要解决的一个重要问题。最后，关于噬菌体制剂的安全性，尚需进行更为详尽且全面的研究。已有研究揭示，噬菌体治疗可能诱导嗜中性粒细胞的更新与功能转变，并在裂解细菌的过程中释放毒素和超级抗原等潜在有害物质，这些物质具有引发包括发热、炎症以及内毒素性休克在内的多种生物学效应的风险^[74]。因此，未来的研究务必充分评估并解决这些潜在的安全风险。

5 展望

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噬菌体凭借其独特的作用机制，成为防治生物被膜相关感染的有效替代疗法。然而，噬菌体疗法依旧存在一些局限性，未来研究仍需聚焦于以下5点：(1) 利用分子生物学领域的前沿技术，从基因层面对噬菌体进行改造(如通过同源重组技术及“共演化-筛选”等方法扩大抗菌谱)，这是当前研究的重要趋势；(2) 进一步研发“噬菌体+”联合治疗方案是抗生物被膜感染的关键策略，如将噬菌体与抗菌肽、中药物质、铁拮抗分子或氯等物质进行组合治疗，从而降低细菌对噬菌体和抗生素形成耐受性的概率；(3) 包被噬菌体以规避中和抗体的作用、延长噬菌体在体内的半衰期，以及去除噬菌体上赋予其免疫原性的蛋白等方法是避免免疫原性影响的重要方向；(4) 未来研究还应重点关注嵌合体内溶素、更新多种给药形式(例如开发气雾剂)、寻求纳米递送载体(如使用藻酸盐-壳聚糖纳米颗粒)及对噬菌体进行基因测序与纯化等途径，从而增强噬菌体及内溶素的临床应用效果与安全性；(5) 针对噬菌体疗法在临床应用中的适用性，未来研究也需要通过更加明确和系统的临床试验来加以验证和评估，以促进噬菌体疗法的安全性评估标准和相关法规的不断完善。

坚信随着科研工作者对噬菌体疗法的不断深入研究和探索，噬菌体将在未来更好地造福于人类健康，并为多个领域的发展贡献其独特价值。

基金

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国家生猪技术创新中心先导科技项目(NCTIP-XD/B12)
国家生猪技术创新中心先导科技项目(NCTIP-XD/C17)
中央高校基本科研业务费(SWU-KQ22045)
重庆市技术创新与应用发展专项面上项目(CSTB2023TIAD-LDX0006)
鲁渝科技协作项目(2022LYXZ030)
云南省科技厅重点研发计划(202403AC100013)

参考文献

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文献

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参考文献引证文献

排序方式：

[1]

刘子豪, 梁媛, 刘丹, 陈红伟, 杨洪早. 根除细菌生物被膜的新视角: 分散[J]. 微生物学报, 2023, 63 (12):4451-4466.

https://doi.org/10.13343/j.cnki.wsxb.20230263

LIU ZH, LIANG Y, LIU D, CHEN HW, YANG HZ. Dispersion: a new perspective for eradicating bacterial biofilm[J]. Acta Microbiologica Sinica, 2023, 63 (12):4451-4466 (in Chinese).

https://doi.org/10.13343/j.cnki.wsxb.20230263

[2]

YAN J, BASSLER BL. Surviving as a community: antibiotic tolerance and persistence in bacterial biofilms[J]. Cell Host & Microbe, 2019, 26 (1):15-21.

[3]

SRIVASTAVA A, VERMA N, KUMAR V, APOORVA P, AGARWAL V. Biofilm inhibition/eradication: exploring strategies and confronting challenges in combatting biofilm[J]. Archives of Microbiology, 2024, 206 (5):212.

https://doi.org/10.1007/s00203-024-03938-0

[4]

李南薇, 刘功良, 高苏娟, 白卫东, 贾爱娟. 生物被膜对食品工业的危害与控制[J]. 食品工业, 2020, 41 (7):317-322.

LI NW, LIU GL, GAO SJ, BAI WD, JIA AJ. Hazards and control of biofilms on the food industry[J]. The Food Industry, 2020, 41 (7):317-322.

[5]

Di DOMENICO EG, OLIVA A, GUEMBE M. The current knowledge on the pathogenesis of tissue and medical device-related biofilm infections[J]. Microorganisms, 2022, 10 (7):1259.

https://doi.org/10.3390/microorganisms10071259

[6]

ZHANG Y, CHENG P, WANG SY, LI XF, PENG LC, FANG RD, XIONG J, LI H, MEI C, GAO JY, SONG ZH, XU DF, FU LZ, LI CH, WU XQ, HE YZ, CHEN HW. Pseudomonas aeruginosa biofilm dispersion by the mouse antimicrobial peptide CRAMP[J]. Veterinary Research, 2022, 53 (1):80.

https://doi.org/10.1186/s13567-022-01097-y

[7]

张阳, 程鹏, 熊静, 李晓芬, 李会, 王士源, 彭练慈, 韩玉竹, 刘娟, 陈红伟. 基于RNA-Seq技术分析鼠源宿主防御肽对铜绿假单胞菌成熟生物被膜的清除作用[J]. 微生物学报, 2021, 61 (9):2843-2853.

https://doi.org/10.13343/j.cnki.wsxb.20200704

ZHANG Y, CHENG P, XIONG J, LI XF, LI H, WANG SY, PENG LC, HAN YZ, LIU J, CHEN HW. Eradication effect of Pseudomonas aeruginosa biofilms by mouse host defense peptide based on RNA-Seq technology[J]. Acta Microbiologica Sinica, 2021, 61 (9):2843-2853 (in Chinese).

https://doi.org/10.13343/j.cnki.wsxb.20200704

[8]

杭柏林, 李雯琪, 曹荔. 治疗用噬菌体的优点、问题与对策[J]. 广东饲料, 2023, 32 (11):35-38.

https://doi.org/10.3969/j.issn.1005-8613.2023.11.012

HANG BL, LI WQ, CAO L. Advantages, problems, and solutions of therapeutic bacteriophages[J]. Guangdong Feed, 2023, 32 (11):35-38 (in Chinese).

https://doi.org/10.3969/j.issn.1005-8613.2023.11.012

[9]

汪宗林, 陈建荣, 尤忠毓, 刘晓侠, 张瑾. 金黄色葡萄球菌生物被膜形成与耐药机制的研究进展[J]. 生物工程学报, 2024, 40 (7):2038-2051.

WANG ZL, CHEN JR, YOU ZY, LIU XX, ZHANG J. Advances in mechanisms of biofilm formation and drug resistance of Staphylococcus aureus[J]. Chinese Journal of Biotechnology, 2024, 40 (7):2038-2051.

[10]

ELOIS MA, SILVA RD, PILATI GVT, RODRÍGUEZ-LÁZARO D, FONGARO G. Bacteriophages as biotechnological tools[J]. Viruses, 2023, 15 (2):349.

https://doi.org/10.3390/v15020349

[11]

DOOLITTLE MM, COONEY JJ, CALDWELL DE. Lytic infection of Escherichia coli biofilms by bacteriophage T4[J]. Canadian Journal of Microbiology, 1995, 41 (1):12-18.

https://doi.org/10.1139/m95-002

[12]

MANOHAR P, LOH B, TURNER D, TAMIZHSELVI R, MATHANKUMAR M, ELANGOVAN N, NACHIMUTHU R, LEPTIHN S. In vitro and in vivo evaluation of the biofilm-degrading Pseudomonas phage Motto, as a candidate for phage therapy[J]. Frontiers in Microbiology, 2024, 15:1344962.

https://doi.org/10.3389/fmicb.2024.1344962

[13]

ZURABOV F, GLAZUNOV E, KOCHETOVA T, USKEVICH V, POPOVA V. Bacteriophages with depolymerase activity in the control of antibiotic resistant Klebsiella pneumoniae biofilms[J]. Scientific Reports, 2023, 13 (1):15188.

https://doi.org/10.1038/s41598-023-42505-3

[14]

LIU BX, GUO QC, LI Z, GUO XX, LIU XC. Bacteriophage endolysin: a powerful weapon to control bacterial biofilms[J]. The Protein Journal, 2023, 42 (5):463-476.

https://doi.org/10.1007/s10930-023-10139-z

[15]

SOONTARACH R, SRIMANOTE P, VORAVUTHIKUNCHAI SP, CHUSRI S. Antibacterial and anti-biofilm efficacy of endolysin LysAB1245 against a panel of important pathogens[J]. Pharmaceuticals, 2024, 17 (2):155.

https://doi.org/10.3390/ph17020155

[16]

NING HQ, LIN H, WANG JX, HE XB, LV XQ, JU L. Characterizations of the endolysin Lys84 and its domains from phage qdsa002 with high activities against Staphylococcus aureus and its biofilms[J]. Enzyme and Microbial Technology, 2021, 148:109809.

https://doi.org/10.1016/j.enzmictec.2021.109809

[17]

FURSOV MV, ABDRAKHMANOVA RO, ANTONOVA NP, VASINA DV, KOLCHANOVA AD, BASHKINA OA, RUBALSKY OV, SAMOTRUEVA MA, POTAPOV VD, MAKAROV VV, YUDIN SM, GINTSBURG AL, TKACHUK AP, GUSHCHIN VA, RUBALSKII EO. Antibiofilm activity of a broad-range recombinant endolysin LysECD7: in vitro and in vivo study[J]. Viruses, 2020, 12 (5):545.

https://doi.org/10.3390/v12050545

[18]

WANG J, LIANG SY, LU XF, XU Q, ZHU Y, YU SY, ZHANG WJ, LIU SG, XIE F. Bacteriophage endolysin Ply113 as a potent antibacterial agent against polymicrobial biofilms formed by enterococci and Staphylococcus aureus[J]. Frontiers in Microbiology, 2023, 14:1304932.

https://doi.org/10.3389/fmicb.2023.1304932

[19]

ŁUSIAK-SZELACHOWSKA M, MIĘDZYBRODZKI R, DRULIS-KAWA Z, CATER K, KNEŽEVIĆ P, WINOGRADOW C, AMARO K, JOŃCZYK-MATYSIAK E, WEBER-DĄBROWSKA B, RĘKAS J, GÓRSKI A. Bacteriophages and antibiotic interactions in clinical practice: what we have learned so far[J]. Journal of Biomedical Science, 2022, 29 (1):23.

https://doi.org/10.1186/s12929-022-00806-1

[20]

MUKHOPADHYAY S, ZHANG PF, TO KKW, LIU YN, BAI CQ, LEUNG SSY. Sequential treatment effects on phage-antibiotic synergistic application against multi-drug-resistant Acinetobacter baumannii[J]. International Journal of Antimicrobial Agents, 2023, 62 (5):106951.

https://doi.org/10.1016/j.ijantimicag.2023.106951

[21]

SOONTARACH R, NWABOR OF, VORAVUTHIKUNCHAI SP. Interaction of lytic phage T1245 with antibiotics for enhancement of antibacterial and anti-biofilm efficacy against multidrug-resistant Acinetobacter baumannii[J]. Biofouling, 2022, 38 (10):994-1005.

https://doi.org/10.1080/08927014.2022.2163479

[22]

胡紫萌, 陈伟叶, 陈欣仪, 李培, 李敏, 周璐, 杜鸿, 刘玉庆, 张炜, 谈忠鸣. PH826噬菌体与抗生素联用有效抑制多重耐药铜绿假单胞菌生物被膜的形成[J]. 微生物学报, 2024, 64 (2):473-488.

https://doi.org/10.13343/j.cnki.wsxb.20230435

HU ZM, CHEN WY, CHEN XY, LI P, LI M, ZHOU L, DU H, LIU YQ, ZHANG W, TAN ZM. Combination of phage PH826 and antibiotics inhibits biofilm formation of multidrug-resistant Pseudomonas aeruginosa[J]. Acta Microbiologica Sinica, 2024, 64 (2):473-488 (in Chinese).

https://doi.org/10.13343/j.cnki.wsxb.20230435

[23]

TAHA M, ARNAUD T, LIGHTLY TJ, PETERS D, WANG LY, CHEN WX, COOK BWM, THERIAULT SS, ABDELBARY H. Combining bacteriophage and vancomycin is efficacious against MRSA biofilm-like aggregates formed in synovial fluid[J]. Frontiers in Medicine, 2023, 10:1134912.

https://doi.org/10.3389/fmed.2023.1134912

[24]

SHREE S, SUMAN E, KOTIAN H, PAUL SH, SUCHITRA SHENOY M. Effect of Klebsiella-specific phage on multidrug-resistant Klebsiella pneumoniae- an experimental study[J]. Indian Journal of Medical Microbiology, 2024, 47:100515.

https://doi.org/10.1016/j.ijmmb.2023.100515

[25]

JDEED G, MOROZOVA V, KOZLOVA Y, TIKUNOV A, USHAKOVA T, BARDASHEVA A, MANAKHOV A, MITINA M, ZHIRAKOVSKAYA E, TIKUNOVA N. StM171, a Stenotrophomonas maltophilia bacteriophage that affects sensitivity to antibiotics in host bacteria and their biofilm formation[J]. Viruses, 2023, 15 (12):2455.

https://doi.org/10.3390/v15122455

[26]

ROSZAK M, DOŁĘGOWSKA B, CECERSKA-HERYĆ E, SERWIN N, JABŁOŃSKA J, GRYGORCEWICZ B. Bacteriophage-Ciprofloxacin combination effectiveness depends on Staphylococcus aureus-Candida albicans dual-species communities' growth model[J]. Microbial Drug Resistance, 2022, 28 (6):613-622.

https://doi.org/10.1089/mdr.2021.0324

[27]

NECEL A, BLOCH S, TOPKA-BIELECKA G, JANISZEWSKA A, ŁUKASIAK A, NEJMAN-FALEŃCZYK B, WĘGRZYN G. Synergistic effects of bacteriophage vB_Eco4-M7 and selected antibiotics on the biofilm formed by shiga Toxin-producing Escherichia coli[J]. Antibiotics, 2022, 11 (6):712.

https://doi.org/10.3390/antibiotics11060712

[28]

HARTMANN R, JECKEL H, JELLI E, SINGH PK, VAIDYA S, BAYER M, RODE DKH, VIDAKOVIC L, DÍAZ-PASCUAL F, FONG JCN, DRAGOŠ A, LAMPRECHT O, THÖMING JG, NETTER N, HÄUSSLER S, NADELL CD, SOURJIK V, KOVÁCS ÁT, YILDIZ FH, DRESCHER K. Quantitative image analysis of microbial communities with BiofilmQ[J]. Nature Microbiology, 2021, 6 (2):151-156.

https://doi.org/10.1038/s41564-020-00817-4

[29]

DRILLING A, MORALES S, JARDELEZA C, VREUGDE S, SPECK P, WORMALD PJ. Bacteriophage reduces biofilm of Staphylococcus aureus ex vivo isolates from chronic rhinosinusitis patients[J]. American Journal of Rhinology & Allergy, 2014, 28 (1):3-11.

[30]

GRYGORCEWICZ B, WOJCIUK B, ROSZAK M, ŁUBOWSKA N, BŁAŻEJCZAK P, JURSA-KULESZA J, RAKOCZY R, MASIUK H, DOŁĘGOWSKA B. Environmental phage-based cocktail and antibiotic combination effects on Acinetobacter baumannii biofilm in a human urine model[J]. Microbial Drug Resistance, 2021, 27 (1):25-35.

https://doi.org/10.1089/mdr.2020.0083

[31]

LUO J, XIE LB, YANG M, LIU M, LI QY, WANG P, FAN JH, JIN J, LUO CH. Synergistic antibacterial effect of phage PB3074 in combination with antibiotics targeting cell wall against Multidrug-resistant Acinetobacter baumannii in vitro and ex vivo[J]. Microbiology Spectrum, 2023, 11 (4):e0034123.

https://doi.org/10.1128/spectrum.00341-23

[32]

ABEDON ST, DANIS-WLODARCZYK KM, WOZNIAK DJ, SULLIVAN MB. Improving phage-biofilm in vitro experimentation[J]. Viruses, 2021, 13 (6):1175.

https://doi.org/10.3390/v13061175

[33]

LUO J, XIE LB, LIU M, LI QY, WANG P, LUO CH. Bactericidal synergism between phage YC#06 and antibiotics: a combination strategy to target multidrug-resistant Acinetobacter baumannii in vitro and in vivo[J]. Microbiology Spectrum, 2022, 10 (4):e0009622.

https://doi.org/10.1128/spectrum.00096-22

[34]

WINTACHAI P, NAKNAEN A, THAMMAPHET J, POMWISED R, PHAONAKROP N, ROYTRAKUL S, SMITH DR. Characterization of extended-spectrum-β-lactamase producing Klebsiella pneumoniae phage KP1801 and evaluation of therapeutic efficacy in vitro and in vivo[J]. Scientific Reports, 2020, 10 (1):11803.

https://doi.org/10.1038/s41598-020-68702-y

[35]

SUNDARAMOORTHY NS, THOTHATHRI S, BHASKARAN M, GANESHPRASAD A, NAGARAJAN S. Phages from Ganges River curtail in vitro biofilms and planktonic growth of drug resistant Klebsiella pneumoniae in a zebrafish infection model[J]. AMB Express, 2021, 11 (1):27.

https://doi.org/10.1186/s13568-021-01181-0

[36]

WANG L, TKHILAISHVILI T, BERNAL ANDRES B, TRAMPUZ A, GONZALEZ MORENO M. Bacteriophage-antibiotic combinations against ciprofloxacin/ceftriaxone-resistant Escherichia coli in vitro and in an experimental Galleria mellonella model[J]. International Journal of Antimicrobial Agents, 2020, 56 (6):106200.

https://doi.org/10.1016/j.ijantimicag.2020.106200

[37]

ABDELGHAFAR A, EL-GANINY A, SHAKER G, ASKOURA M. Isolation of a bacteriophage targeting Pseudomonas aeruginosa and exhibits a promising in vivo efficacy[J]. AMB Express, 2023, 13 (1):79.

https://doi.org/10.1186/s13568-023-01582-3

[38]

KAŹMIERCZAK N, GRYGORCEWICZ B, ROSZAK M, BOCHENTYN B, PIECHOWICZ L. Comparative assessment of bacteriophage and antibiotic activity against multidrug-resistant Staphylococcus aureus biofilms[J]. International Journal of Molecular Sciences, 2022, 23 (3):1274.

https://doi.org/10.3390/ijms23031274

[39]

ALREJA AB, APPEL AE, ZHU JC, RILEY SP, GONZALEZ-JUARBE N, NELSON DC. SP-CHAP, an endolysin with enhanced activity against biofilm pneumococci and nasopharyngeal colonization[J]. mBio, 2024, 15 (4):e0006924.

https://doi.org/10.1128/mbio.00069-24

[40]

VANDER ELST N, BELLEMANS J, LAVIGNE R, BRIERS Y, MEYER E. Endolysin NC5 improves early cloxacillin treatment in a mouse model of Streptococcus uberis mastitis[J]. Applied Microbiology and Biotechnology, 2024, 108 (1):118.

https://doi.org/10.1007/s00253-023-12820-w

[41]

ŁUSIAK-SZELACHOWSKA M, WEBER-DĄBROWSKA B, ŻACZEK M, GÓRSKI A. Anti-biofilm activity of bacteriophages and lysins in chronic rhinosinusitis[J]. Acta Virologica, 2021, 65 (2):127-140.

https://doi.org/10.4149/av_2021_203

[42]

SCHNEIDER G, SZENTES N, HORVÁTH M, DORN Á, COX A, NAGY G, DOFFKAY Z, MARÓTI G, RÁKHELY G, KOVÁCS T. Kinetics of targeted phage rescue in a mouse model of systemic Escherichia coli K1[J]. BioMed Research International, 2018 7569645.

[43]

杨佩霓, 李庆蓉, 李江, 何薇, 和平安, 吕梅, 杨旭. 噬菌体抗细菌生物膜机制及应用策略的研究进展[J]. 现代检验医学杂志, 2024, 39 (1):199-204.

https://doi.org/10.3969/j.issn.1671-7414.2024.01.037

YANG PN, LI QR, LI J, HE W, HE PA, LV M, YANG X. Advances in the mechanism of phage resistance to bacterial biofilms and strategies for its application[J]. Journal of Modern Laboratory Medicine, 2024, 39 (1):199-204 (in Chinese).

https://doi.org/10.3969/j.issn.1671-7414.2024.01.037

[44]

宋军. 金黄色葡萄球菌噬菌体GH15穿孔素功能研究[D]. 长春: 吉林大学博士学位论文, 2016.

SONG J. Study on the function of perforin of Staphylococcus aureus phage GH15[D]. Changchun: Doctoral Dissertation of Jilin University, 2016 (in Chinese).

[45]

CHANG C, YU XB, GUO WN, GUO CY, GUO XK, LI QT, ZHU YZ. Bacteriophage-mediated control of biofilm: a promising new dawn for the future[J]. Frontiers in Microbiology, 2022, 13:825828.

https://doi.org/10.3389/fmicb.2022.825828

[46]

SHARMA U, VIPRA A, CHANNABASAPPA S. Phage-derived lysins as potential agents for eradicating biofilms and persisters[J]. Drug Discovery Today, 2018, 23 (4):848-856.

https://doi.org/10.1016/j.drudis.2018.01.026

[47]

TOPKA-BIELECKA G, DYDECKA A, NECEL A, BLOCH S, NEJMAN-FALEŃCZYK B, WĘGRZYN G, WĘGRZYN A. Bacteriophage-derived depolymerases against bacterial biofilm[J]. Antibiotics, 2021, 10 (2):175.

https://doi.org/10.3390/antibiotics10020175

[48]

AMANKWAH S, ABDELLA K, KASSA T. Bacterial biofilm destruction: a focused review on the recent use of phage-based strategies with other antibiofilm agents[J]. Nanotechnology, Science and Applications, 2021, 14:161-177.

https://doi.org/10.2147/NSA.S325594

[49]

张小妹, 林建涵, 阮士龙, 陈翊平. 控制细菌生物膜感染的可替代策略: 噬菌体疗法[J]. 食品安全质量检测学报, 2023, 14 (21):55-62.

ZHANG XM, LIN JH, RUAN SL, CHEN YP. An alternative strategy to control bacterial biofilm infections: phage therapy[J]. Journal of Food Safety & Quality, 2023, 14 (21):55-62.

[50]

FIGUEIREDO CM, MALVEZZI KARWOWSKI MS, Da SILVA RAMOS RCP, de OLIVEIRA NS, PEÑA LC, CARNEIRO E, FREITAS de MACEDO RE, ROSA EAR. Bacteriophages as tools for biofilm biocontrol in different fields[J]. Biofouling, 2021, 37 (6):689-709.

https://doi.org/10.1080/08927014.2021.1955866

[51]

周彤, 周秀娟, 龙坤兰, 陈骏, 高培阳. 肺炎克雷伯菌生物膜治疗的研究进展[J]. 中国抗生素杂志, 2023, 48 (6):636-642.

https://doi.org/10.3969/j.issn.1001-8689.2023.06.005

ZHOU T, ZHOU XJ, LONG KL, CHEN J, GAO PY. Recent advances in the treatment of Klebsiella pneumoniae biofilms[J]. Chinese Journal of Antibiotics, 2023, 48 (6):636-642 (in Chinese).

https://doi.org/10.3969/j.issn.1001-8689.2023.06.005

[52]

WANG YX, DAI JJ, WANG XH, WANG Y, TANG F. Mechanisms of interactions between bacteria and bacteriophage mediate by quorum sensing systems[J]. Applied Microbiology and Biotechnology, 2022, 106 (7):2299-2310.

https://doi.org/10.1007/s00253-022-11866-6

[53]

van KESSEL JC, MUKHERJEE S. Another battle won in the phage-host arms race: Pseudomonas phage blocks quorum sensing regulator LasR[J]. Molecular Cell, 2021, 81 (3):420-422.

https://doi.org/10.1016/j.molcel.2021.01.007

[54]

DUDDY OP, SILPE JE, FEI CY, BASSLER BL. Natural silencing of quorum-sensing activity protects Vibrio parahaemolyticus from lysis by an autoinducer-detecting phage[J]. PLoS Genetics, 2023, 19 (7):e1010809.

https://doi.org/10.1371/journal.pgen.1010809

[55]

PATEL H, BUCHAD H, GAJJAR D. Pseudomonas aeruginosa persister cell formation upon antibiotic exposure in planktonic and biofilm state[J]. Scientific Reports, 2022, 12 (1):16151.

https://doi.org/10.1038/s41598-022-20323-3

[56]

LIU YP, DAI L, DONG JH, CHEN C, ZHU JG, RAO VB, TAO P. Covalent modifications of the bacteriophage genome confer a degree of resistance to bacterial CRISPR systems[J]. Journal of Virology, 2020, 94 (23):e01630-20.

[57]

SAMSON JE, MAGADÁN AH, SABRI M, MOINEAU S. Revenge of the phages: defeating bacterial defences[J]. Nature Reviews Microbiology, 2013, 11 (10):675-687.

https://doi.org/10.1038/nrmicro3096

[58]

STOKAR-AVIHAIL A, FEDORENKO T, HÖR J, GARB J, LEAVITT A, MILLMAN A, SHULMAN G, WOJTANIA N, MELAMED S, AMITAI G, SOREK R. Discovery of phage determinants that confer sensitivity to bacterial immune systems[J]. Cell, 2023, 186 (9):1863-1876.e16.

https://doi.org/10.1016/j.cell.2023.02.029

[59]

HAMPTON HG, WATSON BNJ, FINERAN PC. The arms race between bacteria and their phage foes[J]. Nature, 2020, 577 (7790):327-336.

https://doi.org/10.1038/s41586-019-1894-8

[60]

高苗, 凌保东. 群体感应介导细菌-噬菌体间相互作用的研究进展[J]. 生物学杂志, 2024. http://kns.cnki.net/kcms/detail/34.1081.Q.20240403.1526.002.html.

GAO M, LING BD. Research progress on quorum sensing mediating bacterial-bacteriophage interactions[J]. Journal of Biology, 2024. http://kns.cnki.net/kcms/detail/34.1081.Q.20240403.1526.002.html (in Chinese).

[61]

WATSON BNJ, STEENS JA, STAALS RHJ, WESTRA ER, van HOUTE S. Coevolution between bacterial CRISPR-Cas systems and their bacteriophages[J]. Cell Host & Microbe, 2021, 29 (5):715-725.

[62]

KEBRIAEI R, LEV KL, SHAH RM, STAMPER KC, HOLGER DJ, MORRISETTE T, KUNZ COYNE AJ, LEHMAN SM, RYBAK MJ. Eradication of biofilm-mediated methicillin-resistant Staphylococcus aureus infections in vitro: bacteriophage-antibiotic combination[J]. Microbiology Spectrum, 2022, 10 (2):e0041122.

https://doi.org/10.1128/spectrum.00411-22

[63]

UCHIYAMA J, SHIGEHISA R, NASUKAWA T, MIZUKAMI K, TAKEMURA-UCHIYAMA I, UJIHARA T, MURAKAMI H, IMANISHI I, NISHIFUJI K, SAKAGUCHI M, MATSUZAKI S. Piperacillin and ceftazidime produce the strongest synergistic phage-antibiotic effect in Pseudomonas aeruginosa[J]. Archives of Virology, 2018, 163 (7):1941-1948.

https://doi.org/10.1007/s00705-018-3811-0

[64]

LIU CX, HONG QX, CHANG RYK, KWOK PCL, CHAN HK. Phage-antibiotic therapy as a promising strategy to combat multidrug-resistant infections and to enhance antimicrobial efficiency[J]. Antibiotics, 2022, 11 (5):570.

https://doi.org/10.3390/antibiotics11050570

[65]

LOGANATHAN A, BOZDOGAN B, MANOHAR P, NACHIMUTHU R. Phage-antibiotic combinations in various treatment modalities to manage MRSA infections[J]. Frontiers in Pharmacology, 2024, 15:1356179.

https://doi.org/10.3389/fphar.2024.1356179

[66]

CHEN Z, GUO ZM, LIN HB, TIAN Y, ZHANG PP, CHEN HS, WANG YW, SHEN YQ. The feasibility of phage therapy for periodontitis[J]. Future Microbiology, 2021, 16:649-656.

https://doi.org/10.2217/fmb-2020-0161

[67]

MIRZAEI A, WAGEMANS J, NASR ESFAHANI B, LAVIGNE R, MOGHIM S. A phage cocktail to control surface colonization by Proteus mirabilis in catheter-associated urinary tract infections[J]. Microbiology Spectrum, 2022, 10 (5):e0209222.

https://doi.org/10.1128/spectrum.02092-22

[68]

WANG CB, HANG H, ZHOU SB, NIU YD, DU HC, STANFORD K, McALLISTER TA. Bacteriophage biocontrol of Shiga toxigenic Escherichia coli (STEC) O145 biofilms on stainless steel reduces the contamination of beef[J]. Food Microbiology, 2020, 92:103572.

https://doi.org/10.1016/j.fm.2020.103572

[69]

SHIVARAM KB, BHATT P, APPLEGATE B, SIMSEK H. Bacteriophage-based biocontrol technology to enhance the efficiency of wastewater treatment and reduce targeted bacterial biofilms[J]. Science of the Total Environment, 2023, 862:160723.

https://doi.org/10.1016/j.scitotenv.2022.160723

[70]

SUN RN, YU PF, ZUO PX, VILLAGRÁN D, MATHIEU J, ALVAREZ PJJ. Biofilm control in flow-through systems using polyvalent phages delivered by peptide-modified M13 coliphages with enhanced polysaccharide affinity[J]. Environmental Science & Technology, 2022, 56 (23):17177-17187.

[71]

LV SX, WANG YH, JIANG KX, GUO XG, ZHANG J, ZHOU F, LI QM, JIANG Y, YANG CY, TENG TS. Genetic engineering and biosynthesis technology: keys to unlocking the chains of phage therapy[J]. Viruses, 2023, 15 (8):1736.

https://doi.org/10.3390/v15081736

[72]

BRIOT T, KOLENDA C, FERRY T, MEDINA M, LAURENT F, LEBOUCHER G, PIROT F. Paving the way for phage therapy using novel drug delivery approaches[J]. Journal of Controlled Release, 2022, 347:414-424.

https://doi.org/10.1016/j.jconrel.2022.05.021

[73]

WROE JA, JOHNSON CT, GARCÍA AJ. Bacteriophage delivering hydrogels reduce biofilm formation in vitro and infection in vivo[J]. Journal of Biomedical Materials Research Part A, 2020, 108 (1):39-49.

https://doi.org/10.1002/jbm.a.36790

[74]

孟祥朋, 孙建和. 防控细菌生物被膜的新武器: 噬菌体[J]. 畜牧与兽医, 2010, 42 (1):97-101.

MENG XP, SUN JH. New insights into the control of bacterial biofilm: bacteriophage[J]. Animal Husbandry & Veterinary Medicine, 2010, 42 (1):97-101.

[75]

ALQAHTANI A. Bacteriophage treatment as an alternative therapy for multidrug-resistant bacteria[J]. Saudi Medical Journal, 2023, 44 (12):1222-1231.

https://doi.org/10.15537/smj.2023.44.12.20230366

[76]

HATFULL GF, DEDRICK RM, SCHOOLEY RT. Phage therapy for antibiotic-resistant bacterial infections[J]. Annual Review of Medicine, 2022, 73:197-211.

https://doi.org/10.1146/annurev-med-080219-122208

2024年第64卷第10期

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doi: 10.13343/j.cnki.wsxb.20240259

接收时间：2024-04-24
首发时间：2026-03-21
出版时间：2024-08-06

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收稿日期：2024-04-24
录用日期：2024-08-02

基金

Pilot Technology Project of National Center of Technology Innovation for Pigs(NCTIP-XD/B12)

国家生猪技术创新中心先导科技项目(NCTIP-XD/B12)

Pilot Technology Project of National Center of Technology Innovation for Pigs(NCTIP-XD/C17)

国家生猪技术创新中心先导科技项目(NCTIP-XD/C17)

Fundamental Research Funds for the Central Universities(SWU-KQ22045)

中央高校基本科研业务费(SWU-KQ22045)

Chongqing Technical Innovation and Application Development Special General Project(CSTB2023TIAD-LDX0006)

重庆市技术创新与应用发展专项面上项目(CSTB2023TIAD-LDX0006)

Science and Technology Cooperation Project of Shandong and Chongqing(2022LYXZ030)

鲁渝科技协作项目(2022LYXZ030)

Yunnan Province Science and Technology Department Key Research and Development Plan(202403AC100013)

云南省科技厅重点研发计划(202403AC100013)

作者信息

1 西南大学动物医学院, 重庆 402460

2 国家生猪技术创新中心, 重庆 402460

3 西南大学医学研究院, 免疫学研究中心, 重庆 402460

4 西南大学, 中兽医药研究所, 重庆 402460

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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Phage	Antibiotic	Pathogenic bacteria	Results
T1245	Ceftazidime, colistin, imipenem, and meropenem	Acinetobacter baumannii	PAC significantly reduced bacterial density by up to 80.0%, disrupted biofilm structure, and altered cell morphology. Additionally, it notably decreased the biomass and bacterial viability of 3-day-old biofilms^[21]
PH826	Ceftazidime and colistin	Pseudomonas aeruginosa	PAC reduced biofilm formation by more than 80.0%^[22]
Remus	Vancomycin	Staphylococcus aureus	In comparison to the use of Remus or vancomycin alone, PAC exhibited greater efficacy in reducing bacterial load. Furthermore, when tested in vivo, PAC achieved the highest survival rate^[23]
Klebsiella-specific phage	Imipenem	Klebsiella pneumoniae	PAC demonstrates a marginal improvement in efficacy compared to phage treatment alone, potentially inhibiting the regrowth of bacteria resistant to the phage to a certain extent^[24]
StM171	Ampicillin, chloramphenicol, levofloxacin, tetracycline, and gentamicin	Stenotrophomonas maltophilia	In most instances, StM171 augmented the inhibitory effect exerted by antibiotics^[25]
vB_SauM-A and vB_SauM-D	Ciprofloxacin	Staphylococcus aureus and Candida albicans	PAC achieved a substantial reduction of 90.0% in biofilm-specific activity for mono-species biofilms and 69.0% for dual-species biofilms, outperforming single phage or antibiotics alone^[26]
vB_Eco4M-7 and ECML-117	Rifampicin and ciprofloxacin	Escherichia coli	PAC effectively reduces the bacterial count and biofilm density. Furthermore, under PAC treatment, the efficiency of emergence of bacteria resistant to phages and rifampicin was significantly diminished^[27]

Phage

Antibiotic

Pathogenic bacteria

Results

T1245

Ceftazidime, colistin, imipenem, and meropenem

Acinetobacter baumannii

PAC significantly reduced bacterial density by up to 80.0%, disrupted biofilm structure, and altered cell morphology. Additionally, it notably decreased the biomass and bacterial viability of 3-day-old biofilms^[21]

PH826

Ceftazidime and colistin

Pseudomonas aeruginosa

PAC reduced biofilm formation by more than 80.0%^[22]

Remus

Vancomycin

Staphylococcus aureus

In comparison to the use of Remus or vancomycin alone, PAC exhibited greater efficacy in reducing bacterial load. Furthermore, when tested in vivo, PAC achieved the highest survival rate^[23]

Klebsiella-specific phage

Imipenem

Klebsiella pneumoniae

PAC demonstrates a marginal improvement in efficacy compared to phage treatment alone, potentially inhibiting the regrowth of bacteria resistant to the phage to a certain extent^[24]

StM171

Ampicillin, chloramphenicol, levofloxacin, tetracycline, and gentamicin

Stenotrophomonas maltophilia

In most instances, StM171 augmented the inhibitory effect exerted by antibiotics^[25]

vB_SauM-A and vB_SauM-D

Ciprofloxacin

Staphylococcus aureus and Candida albicans

PAC achieved a substantial reduction of 90.0% in biofilm-specific activity for mono-species biofilms and 69.0% for dual-species biofilms, outperforming single phage or antibiotics alone^[26]

vB_Eco4M-7 and ECML-117

Rifampicin and ciprofloxacin

Escherichia coli

PAC effectively reduces the bacterial count and biofilm density. Furthermore, under PAC treatment, the efficiency of emergence of bacteria resistant to phages and rifampicin was significantly diminished^[27]

Model	Phage or endolysin	Pathogenic bacteria	Antibiotic	Result
Zebrafish infection model	YC#06	Acinetobacter baumannii	Antibiotic mixtures (chloramphenicol, imipenem, and cefotaxime)	PAC significantly increased the survival rate of zebrafish^[33]
Galleria mellonella larvae survival model	KP1801	Klebsiella pneumoniae	None	Bacteriophage significantly improved the survival rate of larvae. The number of intracellular bacteria in larvae showed a significant decrease while the number of phage increased^[34]
Zebrafish infection model	KpG	Klebsiella pneumoniae	Streptomycin	Treatment of zebrafish muscle tissue with KpG phage and PAC showed a significant 77.7% and 97.2% decline in CFU/ml, respectively^[35]
Galleria mellonella invertebrate infection model	ɸWL-3	Escherichia coli	Fosfomycin	PAC can improve the survival rate of larvae^[36]
Caenorhabditis elegans	Motto	Pseudomonas aeruginosa	None	A 90% survival rate when treated with the phage at a multiplicity of infection of 10.0^[12]
Mice infection model	vB_PaeP_PS28	Pseudomonas aeruginosa	None	After phage treatment, the mortality rate decreased significantly and the bacterial colonization in organs was reduced^[37]
Galleria mellonella model	vB_SauM-D	Staphylococcus aureus	None	Within 60 hours, the control group exhibited 100% lethality, whereas the larvae treated with phage displayed a survival rate of 86% after 120 hours^[38]
Spn mouse nasopharyngeal colonization model	SP-CHAP (endolysin)	Streptococcus pneumoniae	None	The application of SP-CHAP resulted in a notable decrease in the colony-forming units of Spn, surpassing even the effectiveness of Cpl-1^[39]
A mouse model of Streptococcus uberis mastitis	NC5 (VersaTile engineered endolysin)	Streptococcus uberis	Cloxacillin	The high-dose combination therapy significantly reduced the hallmarks of mastitis in fast responders, exhibiting a 13 000-fold reduction in bacterial load, a 5.7-fold decrease in neutrophil infiltration, and a 13.0-fold decrement in the key pro-inflammatory chemokine IL-8^[40]

Model

Phage or endolysin

Pathogenic bacteria

Antibiotic

Result

Zebrafish infection model

YC#06

Acinetobacter baumannii

Antibiotic mixtures (chloramphenicol, imipenem, and cefotaxime)

PAC significantly increased the survival rate of zebrafish^[33]

Galleria mellonella larvae survival model

KP1801

Klebsiella pneumoniae

None

Bacteriophage significantly improved the survival rate of larvae. The number of intracellular bacteria in larvae showed a significant decrease while the number of phage increased^[34]

Zebrafish infection model

KpG

Klebsiella pneumoniae

Streptomycin

Treatment of zebrafish muscle tissue with KpG phage and PAC showed a significant 77.7% and 97.2% decline in CFU/ml, respectively^[35]

Galleria mellonella invertebrate infection model

ɸWL-3

Escherichia coli

Fosfomycin

PAC can improve the survival rate of larvae^[36]

Caenorhabditis elegans

Motto

Pseudomonas aeruginosa

None

A 90% survival rate when treated with the phage at a multiplicity of infection of 10.0^[12]

Mice infection model

vB_PaeP_PS28

Pseudomonas aeruginosa

None

After phage treatment, the mortality rate decreased significantly and the bacterial colonization in organs was reduced^[37]

Galleria mellonella model

vB_SauM-D

Staphylococcus aureus

None

Within 60 hours, the control group exhibited 100% lethality, whereas the larvae treated with phage displayed a survival rate of 86% after 120 hours^[38]

Spn mouse nasopharyngeal colonization model

SP-CHAP (endolysin)

Streptococcus pneumoniae

None

The application of SP-CHAP resulted in a notable decrease in the colony-forming units of Spn, surpassing even the effectiveness of Cpl-1^[39]

A mouse model of Streptococcus uberis mastitis

NC5 (VersaTile engineered endolysin)

Streptococcus uberis

Cloxacillin

The high-dose combination therapy significantly reduced the hallmarks of mastitis in fast responders, exhibiting a 13 000-fold reduction in bacterial load, a 5.7-fold decrease in neutrophil infiltration, and a 13.0-fold decrement in the key pro-inflammatory chemokine IL-8^[40]