Журнал микробиологии, эпидемиологии и иммунобиологии. 2019; : 8-13
Переключение морфотипов колоний Burkholderia pseudomallei при переходе культур в стационарное состояние и в организме экспериментальных животных
https://doi.org/10.36233/0372-9311-2019-2-8-13Аннотация
Цель. Характеристика морфотипов колоний Burkholderia pseudomallei 110, полученных в различных условиях культивирования, изучение их фенотипических свойств и переключения морфологии колоний после снятия стрессового воздействия и в организме экспериментальных животных. Материалы и методы. Морфотипы индуцировали при пассаже B. pseudomallei 110 в LB, стерильной речной воде и клетках Tetrahymena pyriformis, идентифицировали на среде Эшдауна, классифицировали по схеме Chantratita et al. и определяли некоторые фенотипические свойства. Культуры хранили 6-10 месяцев в 0,4% Nutrient agar под вазелиновым маслом и анализировали морфологию колоний. Результаты. Идентифицировали 7 морфотипов колоний, обозначенных I Chl, II, III Chl, IV Chl, V, VI, VII Chl. Вариабельность морфотипов и их соотношение зависели от условий культивирования. Морфотипы отличались активностью внеклеточных ферментов, подвижностью, увеличением продукции поринов, варьированием белковых масс-спектров и снижением вирулентности. От животных при заражении всеми морфотипами выделялись культуры I Chl; при хранении отдельные морфотипы приобретали структуру исходного варианта VI (VII Chl), сходную с ним ферментативную активность и частично восстанавливали вирулентность. Заключение. Морфотип VI (VII Chl) B. pseudomallei 110 в стрессовых условиях дает начало 5 другим морфотипам, которые в организме животных переключаются в морфотип I Chl, а после снятия стрессового воздействия ревертируют в исходный морфологический вариант и восстанавливают его фенотипические свойства.
Список литературы
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5. Cheng A.C., Currie B.J. Melioidosis: epidemiology, pathophysiology and management. Clin. Microbiol. Rev. 2005, 18:383-416.
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7. Currie B. J., Fisher D. A., Howard D. M. et al. Endemic melioidosis in tropical northern Australia: a 10-year prospective study and review of the literature. Clin. Infect. Dis. 2000, 31:981-986.
8. Gierok P., Kohler C., Steinmetz I. et al. Burkholderia pseudomallei colony morphotypes show a synchronized metabolic pattern after acute infection. PLOS Neglected Tropical Diseases. 2016, 10(3): e0004483.
9. Howard K., Inglis T.J. Novel selective medium for isolation of Burkholderia pseudomallei. J. Clin. Microbiol. 2003, 41:3312-3316.
10. Inglis T.J., Sagripanti J.L. Environtmental factors that affect the survival and persistance of Burkholderia pseudomallei. Appl. Environ. Microbiol. 2006, 72:6865-6875.
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12. Tandhavanant S., Thanwisai A., Limmathurotsakul D. et al. Effect of colony morphology variation of Burkholderia pseudomallei on intracellular survival and resistance to antimicrobial environments in human macrophages in vitro. BMC Microbiol. 2010, 10:303.
Journal of microbiology, epidemiology and immunobiology. 2019; : 8-13
Switching of Burkholderia pseudomallei colony morphotypes in stationary condition and in the organism of experimental animals
https://doi.org/10.36233/0372-9311-2019-2-8-13Abstract
Aim. Сharacterization of Burkholderia pseudomallei 110 morphotypes, obtained under various cultivation conditions, studying phenotypic characteristics and switching of colony morphology after removal of stress and in the organism of experimental animals. Materials and methods. Morphotypes were induced by passage of B. pseudomallei 110 in LB, sterile river water and in Tetrahymena pyriformis cells, identified on Ashdown medium, classified according to Chantratita et al., some phenotypic characteristics have been determined. Cultures of morphotypes were stored for 6-10 months in 0,4% Nutrient agar under liquid petrolatun and colony morphology was analyzed. Results. Seven morphotypes of colonies were identified and designated I Chl, II, III Chl, IV Chl, V, VI, and VII Chl. The variability of morphotypes and their ratio depended on cultivation conditions. Morphotypes were distinguished by the activity of extracellular enzymes, mobility, characterized by increase of porin proteins production, variation in protein mass-spectrums, and decrease of virulence. From animals infected with all morphotypes was obtained I Chl morphotype; during storage, all cultures acquired the structure of morphotype VI (VII Chl) of the original strain, similar enzymatic activity and partially restored virulence. Conclusion. The morphotype VI (VII Chl) B. pseudomallei 110 under stress conditions gives rise to 5 other morphotypes that in the animals are switched to the morphotype I Chl; after removal of the stressful effect they are reverted to the initial morphological variant and its phenotypic properties are restored.
References
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2. Ashdown L. R. An improved screening technique for isolation of Pseudomonas pseudomallei from clinical specimens. Pathology. 1979, 11(2):293-297.
3. Chantratita N., Wuthiekanun V., Boonbumrung K. et al. Biological relevance of colony morphology and phenotypic switching by Burkholderia pseudomallei. J. Bacteriol. 2007, 189:807-817.
4. Chen Y. S., Lin H.H., Hung C. C. et al. Phenotypic characteristics and pathogenic ability across distinct morphotypes of Burkholderia pseudomallei DT. Microbiol. Immunol. 2009, 53:184-189.
5. Cheng A.C., Currie B.J. Melioidosis: epidemiology, pathophysiology and management. Clin. Microbiol. Rev. 2005, 18:383-416.
6. Chin C.Y., Hara Y., Ghazali A.K. et al. Global transcriptional analysis of Burkholderia pseudomallei high and low biofilm producers reveals insights into biofilm production and virulence. J. BMC Genomics. 2015, 16:471.
7. Currie B. J., Fisher D. A., Howard D. M. et al. Endemic melioidosis in tropical northern Australia: a 10-year prospective study and review of the literature. Clin. Infect. Dis. 2000, 31:981-986.
8. Gierok P., Kohler C., Steinmetz I. et al. Burkholderia pseudomallei colony morphotypes show a synchronized metabolic pattern after acute infection. PLOS Neglected Tropical Diseases. 2016, 10(3): e0004483.
9. Howard K., Inglis T.J. Novel selective medium for isolation of Burkholderia pseudomallei. J. Clin. Microbiol. 2003, 41:3312-3316.
10. Inglis T.J., Sagripanti J.L. Environtmental factors that affect the survival and persistance of Burkholderia pseudomallei. Appl. Environ. Microbiol. 2006, 72:6865-6875.
11. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970, 227:680-685.
12. Tandhavanant S., Thanwisai A., Limmathurotsakul D. et al. Effect of colony morphology variation of Burkholderia pseudomallei on intracellular survival and resistance to antimicrobial environments in human macrophages in vitro. BMC Microbiol. 2010, 10:303.
