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<article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:ali="http://www.niso.org/schemas/ali/1.0/" article-type="research-article" dtd-version="1.2" xml:lang="en"><front><journal-meta><journal-id journal-id-type="publisher-id">Journal of microbiology, epidemiology and immunobiology</journal-id><journal-title-group><journal-title xml:lang="en">Journal of microbiology, epidemiology and immunobiology</journal-title><trans-title-group xml:lang="ru"><trans-title>Журнал микробиологии, эпидемиологии и иммунобиологии</trans-title></trans-title-group></journal-title-group><issn publication-format="print">0372-9311</issn><issn publication-format="electronic">2686-7613</issn><publisher><publisher-name xml:lang="en">Central Research Institute for Epidemiology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="publisher-id">18821</article-id><article-id pub-id-type="doi">10.36233/0372-9311-690</article-id><article-id pub-id-type="edn">YNTISP</article-id><article-categories><subj-group subj-group-type="toc-heading" xml:lang="en"><subject>ORIGINAL RESEARCHES</subject></subj-group><subj-group subj-group-type="toc-heading" xml:lang="ru"><subject>ОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯ</subject></subj-group><subj-group subj-group-type="article-type"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title xml:lang="en">Stabilization of tigecycline solutions during susceptibility testing of microorganisms by broth microdilution method</article-title><trans-title-group xml:lang="ru"><trans-title>Стабилизация растворов тигециклина при тестировании чувствительности микроорганизмов методом микроразведений в бульоне</trans-title></trans-title-group></title-group><contrib-group><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-4020-0894</contrib-id><name-alternatives><name xml:lang="en"><surname>Kosilova</surname><given-names>Irina S.</given-names></name><name xml:lang="ru"><surname>Косилова</surname><given-names>Ирина Сергеевна</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Biol.), senior researcher, Nutrient medium development laboratory</p></bio><bio xml:lang="ru"><p>канд. биол. наук, c. н. с. лаб. разработки питательных сред</p></bio><email>kosilova.irina@gmail.com</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-4785-6418</contrib-id><name-alternatives><name xml:lang="en"><surname>Domotenko</surname><given-names>Lyubov V.</given-names></name><name xml:lang="ru"><surname>Домотенко</surname><given-names>Любовь Викторовна</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Chem.), leading researcher, Nutrient medium development laboratory</p></bio><bio xml:lang="ru"><p>канд. хим. наук, в. н. с. лаб. разработки питательных сред</p></bio><email>domotenko@obolensk.org</email><xref ref-type="aff" rid="aff1"/></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-4553-3826</contrib-id><name-alternatives><name xml:lang="en"><surname>Khramov</surname><given-names>Mikhail V.</given-names></name><name xml:lang="ru"><surname>Храмов</surname><given-names>Михаил Владимирович</given-names></name></name-alternatives><address><country country="RU">Russian Federation</country></address><bio xml:lang="en"><p>Cand. Sci. (Med.), Deputy director for quality and development</p></bio><bio xml:lang="ru"><p>канд. мед. наук, зам. директора по качеству и развитию</p></bio><email>khramov@obolensk.org</email><xref ref-type="aff" rid="aff1"/></contrib></contrib-group><aff-alternatives id="aff1"><aff><institution xml:lang="en">State Research Center for Applied Microbiology and Biotechnology</institution></aff><aff><institution xml:lang="ru">Государственный научный центр прикладной микробиологии и биотехнологии</institution></aff></aff-alternatives><pub-date date-type="pub" iso-8601-date="2025-09-15" publication-format="electronic"><day>15</day><month>09</month><year>2025</year></pub-date><volume>102</volume><issue>4</issue><issue-title xml:lang="en"/><issue-title xml:lang="ru"/><fpage>474</fpage><lpage>481</lpage><history><date date-type="received" iso-8601-date="2025-05-07"><day>07</day><month>05</month><year>2025</year></date></history><permissions><copyright-statement xml:lang="en">Copyright ©; 2025, Kosilova I.S., Domotenko L.V., Khramov M.V.</copyright-statement><copyright-statement xml:lang="ru">Copyright ©; 2025, Косилова И.С., Домотенко Л.В., Храмов М.В.</copyright-statement><copyright-year>2025</copyright-year><copyright-holder xml:lang="en">Kosilova I.S., Domotenko L.V., Khramov M.V.</copyright-holder><copyright-holder xml:lang="ru">Косилова И.С., Домотенко Л.В., Храмов М.В.</copyright-holder><ali:free_to_read xmlns:ali="http://www.niso.org/schemas/ali/1.0/"/><license><ali:license_ref xmlns:ali="http://www.niso.org/schemas/ali/1.0/">https://creativecommons.org/licenses/by/4.0</ali:license_ref></license></permissions><self-uri xlink:href="https://microbiol.crie.ru/jour/article/view/18821">https://microbiol.crie.ru/jour/article/view/18821</self-uri><abstract xml:lang="en"><p><bold>Introduction.</bold> Tigecycline is widely used in the treatment of infections, leading to the emergence of resistant bacteria. Determining susceptibility by broth microdilution method is recommended to be conducted using freshly prepared Mueller–Hinton broth (MHB) or MHB with antioxidant additives, due to the oxidation of the antibiotic. At the same time, there is no information on the possibility of storing and further using antibiotic solutions.</p> <p><bold>The aim</bold> of the study is to determine the feasibility of stabilizing and rationally using tigecycline solutions to achieve acceptable values of minimum inhibitory concentrations (MIC) when testing control strains by the reference method.</p> <p><bold>Materials and methods.</bold> In the study, the MIC of tigecycline for <italic>Escherichia coli</italic> ATCC 25922, <italic>Staphylococcus aureus</italic> ATCC 29213, <italic>Enterococcus faecalis </italic>ATCC 29212, and <italic>Streptococcus pneumoniae</italic> ATCC 49619 was determined using the microdilution method in MHB, which was pre-prepared and stored for 24–48 hours prior to the study. For the study, a tigecycline stock solution was prepared in water with the addition of various concentrations of Оxyrase, which were stored at 2–6°C and –70°C, and then testing was conducted in accordance with GOST R ISO 20776-1-2022.</p> <p><bold>Results.</bold> The stability of the tigecycline stock solutions does not exceed 5 hours at 2–6°C, but with the addition of 5.0–8.0% Оxyrase, it increases to 16 days, allowing for the determination of MIC values for all test strains within the acceptable range. Changing the storage temperature to –70°C increases the stability of the solutions to 43 days, and with the addition of 5.0–8.0% Оxyrase, to at least 48 weeks</p> <p><bold>Conclusion.</bold> The possibility of stabilizing tigecycline solutions stored at negative temperatures (with and without the addition of Оxyrase) has been determined to obtain acceptable MIC values when determining the susceptibility of control strains to antimicrobial agents. The use of these solutions allowed for a reduction in testing costs through the rational use of the antibiotic.</p></abstract><trans-abstract xml:lang="ru"><p><bold>Введение.</bold> Тигециклин широко используется при лечении инфекций, что приводит к появлению устойчивых к нему бактерий. Определение чувствительности методом микроразведений в бульоне рекомендовано проводить, используя свежеприготовленный бульон Мюллера–Хинтон (МХБ) или МХБ с антиоксидантными добавками, что обусловлено окислением антибиотика. Вместе с тем отсутствует информация о возможности хранения и дальнейшего использования растворов антибиотика.</p> <p><bold>Цель </bold>исследования <bold>— </bold>определить возможность стабилизации и рационального использования растворов тигециклина для получения допустимых значений минимальных подавляющих концентраций (МПК) при тестировании контрольных штаммов референтным методом.</p> <p><bold>Материалы и методы.</bold> В работе определяли МПК тигециклина для <italic>Escherichia соli</italic> АТСС 25922, <italic>Staphylococcus aureus</italic> АТСС 29213, <italic>Enterococcus faecalis</italic> ATCC 29212, <italic>Streptococcus pneumoniae</italic> ATCC 49619 методом микроразведений в МХБ, заранее приготовленном и хранившемся 24–48 ч до исследования. Для исследования готовили базовый раствор тигециклина в воде и с добавлением различных концентраций оксиразы, которые хранили при 2–6°С и –70°С и затем проводили тестирование в соответствии с ГОСТ Р ИСО 20776-1-2022.</p> <p><bold>Результаты.</bold> В ходе исследования установлено, что при 2–6°С стабильность базовых растворов тигециклина сохраняется не более 5 ч, а с добавлением 5,0–8,0% оксиразы увеличивается до 16 сут, что позволяет получать значения МПК для всех тест-штаммов в допустимом диапазоне. Изменение температуры хранения до –70°С увеличивает стабильность растворов до 43 сут, а с добавлением 5,0–8,0% оксиразы — минимум до 48 нед.</p> <p><bold>Заключение.</bold> Определена возможность стабилизации растворов тигециклина, хранившихся при отрицательных температурах (с добавлением оксиразы и без неё) для получения допустимых значений МПК при определении чувствительности контрольных штаммов к антимикробным препаратам. Применение данных растворов позволило снизить затраты на тестирование за счёт рационального использования антибиотика.</p></trans-abstract><kwd-group xml:lang="en"><kwd>tigecycline</kwd><kwd>susceptibility testing</kwd><kwd>broth microdilution method</kwd><kwd>Оxyrase</kwd><kwd>Mueller–Hinton broth</kwd></kwd-group><kwd-group xml:lang="ru"><kwd>тигециклин</kwd><kwd>определение чувствительности</kwd><kwd>метод микроразведений в бульоне</kwd><kwd>оксираза</kwd><kwd>бульон Мюллера–Хинтон</kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution xml:lang="en">Federal Service for the Oversight of Consumer Protection and Welfare of the Russian Federation</institution></institution-wrap><institution-wrap><institution xml:lang="ru">Федеральная служба по надзору в сфере защиты прав потребителей и благополучия человека</institution></institution-wrap></funding-source></award-group></funding-group></article-meta></front><body></body><back><ref-list><ref id="B1"><label>1.</label><mixed-citation>Laxminarayan R., Sridhar D., Blaser M., et al. Achieving global targets for antimicrobial resistance. Science. 2016; 353(6302): 874–9. DOI: https://doi.org/10.1126/science.aaf9286</mixed-citation></ref><ref id="B2"><label>2.</label><mixed-citation>Karageorgopoulos D.E., Falagas M.E. Current control and treatment of multidrug-resistant Acinetobacter baumannii infections. Lancet Infect. Dis. 2008;8(12):751–62. DOI: https://doi.org/10.1016/s1473-3099(08)70279-2</mixed-citation></ref><ref id="B3"><label>3.</label><mixed-citation>Rodríguez-Baño J., Gutiérrez-Gutiérrez B., Machuca I., et al. Treatment of infections caused by extended-spectrum-beta-lactamase-, AmpC-, and carbapenemase-producing Enterobacteriaceae. Clin. Microbiol. Rev. 2018;31(2):e00079-17. DOI: https://doi.org/10.1128/cmr.00079-17</mixed-citation></ref><ref id="B4"><label>4.</label><mixed-citation>IACG Discussion Paper. Interagency Coordination Group on Antimicrobial Resistance. Reduce unintentional exposure and the need for antimicrobials, and optimize their use;2018.</mixed-citation></ref><ref id="B5"><label>5.</label><mixed-citation>Шедько Е.Д., Тимошина О.Ю., Азизов И.С. Молекулярная эпидемиология генов группы mcr. Клиническая микробиология и антимикробная химиотерапия. 2020;22(4): 287–300. Shedko E.D., Timoshina O.Yu., Azizov I.S. Molecular epidemiology of mcr group genes. Clinical Microbiology and Antimicrobial Chemotherapy. 2020;22(4):287–300. DOI: https://doi.org/10.36488/cmac.2020.4.287-300 EDN: https://elibrary.ru/ycsxgu</mixed-citation></ref><ref id="B6"><label>6.</label><mixed-citation>Partridge S.R., Pilato V.D., Doi Y., et al. Proposal for assignment of allele numbers for mobile colistin resistance (mcr) genes. J. Antimicrob. Chemother. 2018;73(10):2625–30. DOI: https://doi.org/10.1093/jac/dky262</mixed-citation></ref><ref id="B7"><label>7.</label><mixed-citation>Cheng Y., Li Y., Yu R., et al. Identification of novel tet(X3) variants resistant to tigecycline in Acinetobacter species. Microbiol. Spectr. 2022;10(6):e0133322. DOI: https://doi.org/10.1128/spectrum.01333-22</mixed-citation></ref><ref id="B8"><label>8.</label><mixed-citation>Tasina E., Haidich A.B., Kokkali S., et al. Efficacy and safety of tigecycline for the treatment of infectious diseases: a meta-analysis. Lancet Infect. Dis. 2011;11(11): 834–44. DOI: https://doi.org/10.1016/s1473-3099(11)70177-3</mixed-citation></ref><ref id="B9"><label>9.</label><mixed-citation>Korczak L., Majewski P., Iwaniuk D., et al. Molecular mechanisms of tigecycline-resistance among Enterobacterales. Front. Cell. Infect. Microbiol. 2024;14:1289396. DOI: https://doi.org/10.3389/fcimb.2024.1289396</mixed-citation></ref><ref id="B10"><label>10.</label><mixed-citation>He T., Wang R., Liu D., et al. Emergence of plasmid-mediated high-level tigecycline resistance genes in animals and humans. Nat. Microbiol. 2019;4(9):1450–6. DOI: https://doi.org/10.1038/s41564-019-0445-2</mixed-citation></ref><ref id="B11"><label>11.</label><mixed-citation>Liu C., Liu J., Lu Q., et al. The mechanism of tigecycline resistance in Acinetobacter baumannii under sub-minimal inhibitory concentrations of tigecycline. Int. J. Mol. Sci. 2024;25(3):1819. DOI: https://doi.org/10.3390/ijms25031819</mixed-citation></ref><ref id="B12"><label>12.</label><mixed-citation>Bradford P.A., Petersen P.J., Young M., et al. Tigecycline MIC testing by broth dilution requires use of fresh medium or addition of the biocatalytic oxygen-reducing reagent Oxyrase to standardize the test method. Antimicrob. Agents Chemother. 2005;49(9):3903–9. DOI: https://doi.org/10.1128/aac.49.9.3903-3909.2005</mixed-citation></ref><ref id="B13"><label>13.</label><mixed-citation>Brown S.D., Traczewski M.M. Comparative in vitro antimicrobial activity of tigecycline, a new glycylcycline compound, in freshly prepared medium and quality control. J. Clin. Microbiol. 2007;45(7):2173–9. DOI: https://doi.org/10.1128/jcm.02351-06</mixed-citation></ref><ref id="B14"><label>14.</label><mixed-citation>Petersen P.J., Bradford P.A. Effect of medium age and supplementation with the biocatalytic oxygen-reducing reagent Oxyrase on in vitro activities of tigecycline against recent clinical isolates. Antimicrob. Agents Chemother. 2005;49(9):3910–8. DOI: https://doi.org/10.1128/aac.49.9.3910-3918.2005</mixed-citation></ref><ref id="B15"><label>15.</label><mixed-citation>Jitkova Y., Gronda M., Hurren R., et al. A novel formulation of tigecycline has enhanced stability and sustained antibacterial and antileukemic activity. PLoS One. 2014;9(5):e95281. DOI: https://doi.org/10.1371/journal.pone.0095281</mixed-citation></ref><ref id="B16"><label>16.</label><mixed-citation>Amann L.F., Vicente E.R., Rathke M., et al. Stability studies with tigecycline in bacterial growth medium and impact of stabilizing agents. Eur. J. Clin. Microbiol. Infect. Dis. 2021;40(1):215–8. DOI: https://doi.org/10.1007/s10096-020-03970-0</mixed-citation></ref><ref id="B17"><label>17.</label><mixed-citation>Zhou H., Sun X., Lyu S., et al. Evaluation of tigecycline utilization and trends in antibacterial resistance from 2018 to 2021 in a comprehensive teaching hospital in China. Infect. Drug Resist. 2023;16:879–89. DOI: https://doi.org/10.2147/idr.s395158</mixed-citation></ref></ref-list></back></article>
