{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T13:19:50Z","timestamp":1753881590932,"version":"3.41.2"},"reference-count":53,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2025,2,4]],"date-time":"2025-02-04T00:00:00Z","timestamp":1738627200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Microbiol Spectr"],"published-print":{"date-parts":[[2025,2,4]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:sec>\n            <jats:title\/>\n            <jats:p>\n              Mycoplasma (Class: Mollicutes) contamination in cell cultures is a universal concern for research laboratories. Some estimates report contamination in up to 35% of continuous cell lines. Various commercial antibiotic treatments can successfully decontaminate clean cell lines\n              <jats:italic>in vitro<\/jats:italic>\n              ; however,\n              <jats:italic>in vitro<\/jats:italic>\n              decontamination of bacterial cultures remains challenging. Intracellular bacteria like those in the genera\n              <jats:italic>Rickettsia<\/jats:italic>\n              and\n              <jats:italic>Ehrlichia<\/jats:italic>\n              require cell culture for primary isolation and propagation and are thus vulnerable to contamination with mycoplasmas. Some analyses have reported successful antibiotic clearance of contaminating mycoplasmas in\n              <jats:italic>Rickettsia<\/jats:italic>\n              cultures; however, many of these studies do not identify the contaminating mycoplasma species and often include only a few isolates. To our knowledge, there are no published studies reporting decontamination of mycoplasmas from\n              <jats:italic>Ehrlichia<\/jats:italic>\n              cultures. In this study, we developed a specific multiplex assay to identify two of the most common mycoplasma culture contaminants,\n              <jats:italic>Mycoplasma arginini<\/jats:italic>\n              and\n              <jats:italic>Mycoplasma orale<\/jats:italic>\n              , in cell cultures infected with\n              <jats:italic>Rickettsia<\/jats:italic>\n              or\n              <jats:italic>Ehrlichia<\/jats:italic>\n              species. We further describe the successful\n              <jats:italic>in vitro<\/jats:italic>\n              decontamination of\n              <jats:italic>M. arginini<\/jats:italic>\n              ,\n              <jats:italic>M. orale,<\/jats:italic>\n              and co-contaminations with both mycoplasmas from multiple\n              <jats:italic>Rickettsia<\/jats:italic>\n              and\n              <jats:italic>Ehrlichia<\/jats:italic>\n              cultures using daptomycin and clindamycin.\n            <\/jats:p>\n            <jats:sec>\n              <jats:title>IMPORTANCE<\/jats:title>\n              <jats:p>\n                Mycoplasma contamination is a frequent problem in bacterial cell culture. These prolific organisms thrive in the extracellular environment\n                <jats:italic>in vitro<\/jats:italic>\n                and can persist in cell lines indefinitely without treatment. Historically, mycoplasma-contaminated\n                <jats:italic>Rickettsia<\/jats:italic>\n                cultures were cleared of contaminants by inoculating laboratory mice and re-isolating mycoplasma-free\n                <jats:italic>Rickettsia<\/jats:italic>\n                from brain endothelial cells. However, this method requires the sacrifice of live animals and is not always effective. Mycoplasma clearance via mouse inoculation requires a patent infection of murine central nervous system endothelial cells, which may not occur with some mildly pathogenic or nonpathogenic rickettsial species.\n                <jats:italic>In vitro<\/jats:italic>\n                antibiotic treatment represents an alternate method to eliminate contaminating mycoplasmas from rickettsial cultures. This method requires minimal adjustment of laboratories that already maintain rickettsial cultures and is not dependent on the use of laboratory animals. As such, the comprehensive strategy for\n                <jats:italic>Mycoplasma arginini<\/jats:italic>\n                and\n                <jats:italic>Mycoplasma orale<\/jats:italic>\n                elimination presented here can improve laboratory efficiency for\n                <jats:italic>in vitro<\/jats:italic>\n                research with intracellular bacteria.\n              <\/jats:p>\n            <\/jats:sec>\n          <\/jats:sec>","DOI":"10.1128\/spectrum.01743-24","type":"journal-article","created":{"date-parts":[[2025,1,16]],"date-time":"2025-01-16T14:01:28Z","timestamp":1737036088000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Molecular identification and antibiotic clearance of\n            <i>Mycoplasma arginini<\/i>\n            and\n            <i>Mycoplasma orale<\/i>\n            from cell cultures infected with\n            <i>Rickettsia<\/i>\n            or\n            <i>Ehrlichia<\/i>\n            species"],"prefix":"10.1128","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2758-3245","authenticated-orcid":true,"given":"Michelle E. J.","family":"Allerdice","sequence":"first","affiliation":[{"name":"Division of Vector-borne Diseases, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Savannah L.","family":"Shooter","sequence":"additional","affiliation":[{"name":"Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6823-0185","authenticated-orcid":true,"given":"Maria F. B. M.","family":"Galletti","sequence":"additional","affiliation":[{"name":"Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2786-0057","authenticated-orcid":false,"given":"Joy A.","family":"Hecht","sequence":"additional","affiliation":[{"name":"Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sandor E.","family":"Karpathy","sequence":"additional","affiliation":[{"name":"Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1615-8526","authenticated-orcid":false,"given":"Christopher D.","family":"Paddock","sequence":"additional","affiliation":[{"name":"Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services","place":["Atlanta, USA"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2010.43"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.32607\/20758251-2014-6-3-41-51"},{"key":"e_1_3_3_4_2","volume-title":"Medical microbiolgy","author":"Razin S","year":"1996","unstructured":"Razin S. 1996. 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