{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T16:32:25Z","timestamp":1791995545822,"version":"4.4.0"},"reference-count":29,"publisher":"American Society for Microbiology","issue":"11","license":[{"start":{"date-parts":[[2011,6,1]],"date-time":"2011-06-01T00:00:00Z","timestamp":1306886400000},"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":["Appl Environ Microbiol"],"published-print":{"date-parts":[[2011,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    We recently described acidified citrate cysteine medium (ACCM), which supports host cell-free (axenic) growth of\n                    <jats:named-content content-type=\"genus-species\">Coxiella burnetii<\/jats:named-content>\n                    . After 6 days of incubation, greater than 3 logs of growth was achieved with the avirulent Nine Mile phase II (NMII) strain. Here, we describe modified ACCM and culture conditions that support improved growth of\n                    <jats:named-content content-type=\"genus-species\">C. burnetii<\/jats:named-content>\n                    and their use in genetic transformation and pathogen isolation from tissue samples. ACCM was modified by replacing fetal bovine serum with methyl-\u03b2-cyclodextrin to generate ACCM-2. Cultivation of NMII in ACCM-2 with moderate shaking and in 2.5% oxygen yielded 4 to 5 logs of growth over 7 days. Similar growth was achieved with the virulent Nine Mile phase I and G isolates of\n                    <jats:named-content content-type=\"genus-species\">C. burnetii<\/jats:named-content>\n                    . Colonies that developed after 6 days of growth in ACCM-2 agarose were approximately 0.5 mm in diameter, roughly 5-fold larger than those formed in ACCM agarose. By electron microscopy, colonies consisted primarily of the\n                    <jats:named-content content-type=\"genus-species\">C. burnetii<\/jats:named-content>\n                    small cell variant morphological form. NMII was successfully cultured in ACCM-2 when medium was inoculated with as little as 10 genome equivalents contained in tissue homogenates from infected SCID mice. A completely axenic\n                    <jats:named-content content-type=\"genus-species\">C. burnetii<\/jats:named-content>\n                    genetic transformation system was developed using ACCM-2 that allowed isolation of transformants in about 2 1\/2 weeks. Transformation experiments demonstrated clonal populations in colonies and a transformation frequency of approximately 5 \u00d7 10\n                    <jats:sup>\u22125<\/jats:sup>\n                    . Cultivation in ACCM-2 will accelerate development of\n                    <jats:named-content content-type=\"genus-species\">C. burnetii<\/jats:named-content>\n                    genetic tools and provide a sensitive means of primary isolation of the pathogen from Q fever patients.\n                  <\/jats:p>","DOI":"10.1128\/aem.02826-10","type":"journal-article","created":{"date-parts":[[2011,4,8]],"date-time":"2011-04-08T22:35:10Z","timestamp":1302302110000},"page":"3720-3725","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":225,"title":["Isolation from Animal Tissue and Genetic Transformation of Coxiella burnetii Are Facilitated by an Improved Axenic Growth Medium"],"prefix":"10.1128","volume":"77","author":[{"given":"Anders","family":"Omsland","sequence":"first","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul A.","family":"Beare","sequence":"additional","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joshua","family":"Hill","sequence":"additional","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"},{"name":"Department of Microbial and Molecular Pathogenesis, Texas A&amp;M Health Sciences Center, College Station, Texas 77843"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diane C.","family":"Cockrell","sequence":"additional","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dale","family":"Howe","sequence":"additional","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bryan","family":"Hansen","sequence":"additional","affiliation":[{"name":"Electron Microscopy Unit, Research Technologies Branch, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James E.","family":"Samuel","sequence":"additional","affiliation":[{"name":"Department of Microbial and Molecular Pathogenesis, Texas A&amp;M Health Sciences Center, College Station, Texas 77843"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert A.","family":"Heinzen","sequence":"additional","affiliation":[{"name":"Coxiella Pathogenesis Section, Laboratory of Intracellular Parasites"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1196\/annals.1355.026"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.2005.04539.x"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01580-08"},{"key":"e_1_3_2_5_2","volume-title":"Abstr. 24th Meet. 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