{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T06:22:17Z","timestamp":1783059737025,"version":"3.54.6"},"reference-count":59,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:00:00Z","timestamp":1783036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Bovine mastitis is a prevalent infectious disease in dairy herds worldwide, resulting in substantial economic losses.\n                      <jats:italic>Citrobacter freundii<\/jats:italic>\n                      has emerged as an opportunistic pathogen associated with mastitis, and its increasing antibiotic resistance poses significant therapeutic challenges.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      In this study, a novel lytic phage, Citro-6, was isolated from dairy farm sewage and fecal samples using\n                      <jats:italic>C. freundii<\/jats:italic>\n                      Z6 as the host bacterium. The phage was characterized through morphological and genomic analyses. Its host range, stability under varying temperature and pH conditions, and effects on bacterial adhesion, invasion, cytotoxicity, and pro-inflammatory cytokine expression in bovine mammary epithelial cells were evaluated in vitro.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      Morphological and genomic analyses identified Citro-6 as a member of the class Caudoviricetes. It encodes a complete endolysin sequence with an intact catalytic domain and lacks virulence or antibiotic resistance genes. Citro-6 exhibited broad lytic activity, lysing 83.3% of tested\n                      <jats:italic>Citrobacter<\/jats:italic>\n                      strains, and remained stable at temperatures up to 45\u202f\u00b0C and across a pH range of 5.0 to 9.0. In cellular assays, Citro-6 treatment significantly reduced bacterial adhesion and invasion by approximately 1.66 and 1.19 log units, respectively. Furthermore, it effectively attenuated pathogen-induced cytotoxicity, reducing cell death by approximately 75.2% at 8 h post-infection compared with the Z6-infected group. Citro-6 also prevented morphological cellular damage and suppressed the overexpression of pro-inflammatory cytokines, resulting in decreases in IL-1\u03b2 and TNF-\n                      <jats:italic>\u03b1<\/jats:italic>\n                      concentrations of approximately 37.1% and 47.6%, respectively, relative to the infected control.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      These findings demonstrate that phage Citro-6 is a promising biocontrol candidate against\n                      <jats:italic>C. freundii<\/jats:italic>\n                      -induced bovine mastitis; however, further\n                      <jats:italic>in vivo<\/jats:italic>\n                      evaluation is warranted to confirm its therapeutic potential.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fmicb.2026.1879434","type":"journal-article","created":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T05:41:26Z","timestamp":1783057286000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Isolation, characterization, and in vitro efficacy of phage Citro-6 against Citrobacter freundii-induced bovine mastitis"],"prefix":"10.3389","volume":"17","author":[{"given":"Ziyi","family":"Wang","sequence":"first","affiliation":[{"name":"School of Life Sciences, Inner Mongolia University","place":["Hohhot, Inner Mongolia, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hong","family":"Li","sequence":"additional","affiliation":[{"name":"Ejin Horo Animal Disease Prevention and Control Center","place":["Ordos, Inner Mongolia, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rihua","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Life Sciences, Inner Mongolia University","place":["Hohhot, Inner Mongolia, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ke","family":"Meng","sequence":"additional","affiliation":[{"name":"Ejin Horo Animal Disease Prevention and Control Center","place":["Ordos, Inner Mongolia, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Zhu","sequence":"additional","affiliation":[{"name":"School of Physical Science and Technology, Inner Mongolia University","place":["Hohhot, Inner Mongolia, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,7,3]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"635597","DOI":"10.3389\/fcimb.2021.635597","article-title":"Rekindling of a masterful precedent; bacteriophage: reappraisal and future pursuits","volume":"11","author":"Abd-Allah","year":"2021","journal-title":"Front. 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