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This is not only a huge burden on the global economy but also a growing threat to animal, environmental, plant, and human health, and new strategies are needed to avoid resistance and improve treatment. Novel antimicrobial resistance genes are normally first detected once they cause problems in clinical infections, and we have limited knowledge on their evolutionary trajectories. Current antimicrobial susceptibility testing and research have a limited focus on key environmental factors in pathogen-reservoir-host interactions, possibly leading to inaccurate results that do not reflect the\n                      <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                      conditions. Focusing on differences in pH, we determined the MIC of a panel of isogenic strains expressing CTX-M-15 and CMY-2 \u03b2-lactamases. We found that pH has a large impact on the activity of \u03b2-lactamases, and treatment of these resistant isolates could be possible if the pH of the environment is modified. We verified this using enzyme kinetics, co-cultures, and growth experiments, suggesting that exposure to different environmental conditions may lead to distinct evolutionary trajectories for specific \u03b2-lactamases. Exploring the effect of different temperatures, we also observed a differential effect of avian and mammal host temperatures. Environmental factors such as pH and temperature may have a large unnoticed effect on antimicrobial resistance, and we might use this knowledge to renew and extend the use of old antibiotics for certain infections.\n                    <\/jats:p>\n                    <jats:sec>\n                      <jats:title>IMPORTANCE<\/jats:title>\n                      <jats:p>Antimicrobial resistance is a huge burden to global health and economy. We need new options for avoiding selection of resistance and improved treatment. Overlooked aspect: current susceptibility testing does not take pH into account. With this study, we show that pH and temperature can have large and contrasting effects on the activity (and therefore MIC) of specific \u03b2-lactamases. This might help to explain the phenomenon of bacteria often harboring multiple \u03b2-lactamases seemingly with the same function as well as be utilized to enable treatment of genotypically resistant strains under very specific conditions, that is, treatment of CTX-M-15, the most prevalent ESBL in healthcare, under alkaline conditions.<\/jats:p>\n                    <\/jats:sec>\n                  <\/jats:sec>","DOI":"10.1128\/aem.01775-25","type":"journal-article","created":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T05:00:19Z","timestamp":1766984419000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":3,"title":["Contrasting pH optima of \u03b2-lactamases CTX-M and CMY influence\n                    <i>Escherichia coli<\/i>\n                    fitness and resistance ecology"],"prefix":"10.1128","volume":"92","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0685-1055","authenticated-orcid":false,"given":"Mikkel","family":"Anbo","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2538-8086","authenticated-orcid":true,"given":"Saria","family":"Otani","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0077-5207","authenticated-orcid":true,"given":"Mirena","family":"Ivanova","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanne N\u00f8rgaard","family":"Nielsen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jacob Dyring","family":"Jensen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christina Aaby","family":"Svendsen","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chengfang","family":"Pang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7116-2723","authenticated-orcid":true,"given":"Frank M.","family":"Aarestrup","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04qtj9h94","id-type":"ROR","asserted-by":"publisher"}],"name":"DTU National Food Institute, Technical University of Denmark","place":["Lyngby, Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_4_2_2","unstructured":"FAO UNEP WHO WOAH. 2022. 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