Śliwka Paulina, Moreno David Sáez, Korzeniowski Paweł, Milcarz Agata, Kuczkowski Maciej, Kolenda Rafał, Kozioł Sylwia, Narajczyk Magdalena, Roesler Uwe, Tomaszewska-Hetman Ludwika, Kuźmińska-Bajor Marta
Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Sciences, Wrocław University of Environmental and Life Sciences, Poland.
CEB-Centre of Biological Engineering, University of Minho, Braga 4710-057, Portugal.
Vet Microbiol. 2025 Feb;301:110363. doi: 10.1016/j.vetmic.2024.110363. Epub 2025 Jan 2.
Avian pathogenic Escherichia coli (APEC) is a principal etiologic agent of avian colibacillosis, responsible for significant economic losses in the poultry industry due to high mortality and disease treatment with antibiotics. APEC and its ability to form biofilms on food and processing surfaces contributes to its persistence within farms. Bacteriophages are promising antibacterial agents for combating APEC. This study focused on characterization of the newly isolated phages UPWr_E1, UPWr_E2, and UPWr_E4 as well as the UPWr_E124 phage cocktail containing these three phages. Methods included efficiency of plating assay, transmission electron microscopy, and characterization of their resistance to different pH values and temperatures. Moreover, phage genomes were sequenced, annotated and analyzed, and were compared with previously sequenced E. coli phages. All three phages are virulent and devoid of undesirable genes for therapy. Phage UPWr_E1 belongs to the genus Krischvirus within the order Straboviridae and both UPWr_E2 and UPWr_E4 belong to the genus Tequatrovirus within the subfamily Tevenvirinae, sharing over 95 % nucleotide identity between them. For their use on poultry farms, UPWr_E phages and the UPWr_E124 phage cocktail were tested for their anti-biofilm activity on two E. coli strains - 158B (APEC) and the strong biofilm producer NCTC 17848 - on two abiotic surfaces: a 96-well microplate, a stainless steel surface, and one biotic surface, represented by lettuce leaves. The reduction of biofilm formed by both strains in the 96-well microplate, on the stainless steel and lettuce leaf surface for bacteriophage treatment was very efficient, reducing biofilms by ranges of 50.2-83.6, 58.2-88.4 and 53-99.4 %, respectively. Therefore, we conclude that UPWr_E phages and the UPWr_E124 phage cocktail are promising candidates for APEC biocontrol.
禽致病性大肠杆菌（APEC）是禽大肠杆菌病的主要病原体，因其高死亡率和抗生素疾病治疗导致家禽业重大经济损失。APEC及其在食品和加工表面形成生物膜的能力有助于其在养殖场内持续存在。噬菌体是对抗APEC的有前景的抗菌剂。本研究重点对新分离的噬菌体UPWr_E1、UPWr_E2和UPWr_E4以及包含这三种噬菌体的UPWr_E124噬菌体鸡尾酒进行特性分析。方法包括平板效率测定、透射电子显微镜以及对其不同pH值和温度抗性的表征。此外，对噬菌体基因组进行测序、注释和分析，并与先前测序的大肠杆菌噬菌体进行比较。所有三种噬菌体均为烈性噬菌体且无用于治疗的不良基因。噬菌体UPWr_E1属于Straboviridae目Krischvirus属，UPWr_E2和UPWr_E4均属于Tevenvirinae亚科Tequatrovirus属，它们之间共享超过95%的核苷酸同一性。为了在养禽场使用，对UPWr_E噬菌体和UPWr_E124噬菌体鸡尾酒在两种大肠杆菌菌株——158B（APEC）和强生物膜产生菌NCTC 17848——在两种非生物表面：96孔微孔板、不锈钢表面以及一种生物表面（以生菜叶为代表）上的抗生物膜活性进行了测试。在96孔微孔板、不锈钢和生菜叶表面，噬菌体处理对两种菌株形成的生物膜的减少非常有效，分别将生物膜减少了50.2 - 83.6%、58.2 - 88.4%和53 - 99.4%。因此，我们得出结论，UPWr_E噬菌体和UPWr_E124噬菌体鸡尾酒是APEC生物防治的有前景的候选物。
