Activation of the xenobiotics biotransformation of E. coii with development of resistance to Cefotaxime and Apramycin and their composition

Prolonged use of antibiotics of one class leads to the formation of stable populations of infectious agents, the etiology of which is often due to resistant forms and types of pathogenic and opportunistic microorganisms. Factors contributing to the emergence of antibacterial resistance are the cyclic use of antibiotics of one class, the use of nonspecific, low-performance antibiotics, the choice of the drug without taking into account the localization of the inflammatory process and species of the causative agent, failure to comply with the dosage and duration of the course of treatment, etc. To demonstrate the bactericidal effect of the antimicrobial drug against resistant strains of microorganisms, the concentration of the active substance is higher than therapeutics The main reason for the formation of bacterial resistance to many antimicrobial drugs used is the preparation of R-plasmids conjugated from other bacteria carrying antibiotic resistance genes that are capable of transferring from one bacterium to another, forming resistance in the whole population. A promising direction in solving the problem of reducing the formation of resistance in bacteria is the creation of complex preparations containing components with different mechanisms and spectrum of antimicrobial action. Laboratory studies were conducted on various clones of the E. coli strain 866, resistant to previously developed complex antimicrobial agents and active ingredients that make up them. After every ten passages, the method of serial dilutions was used to study the antimicrobial activity of the preparations with respect to these microorganisms. Isolation of plasmid DNA from bacterial cells was performed by alkaline lysis and subsequent binding on selective membranes of microspin columns using a plasmid DNA extraction kit. Isolation of total cellular RNA was carried out by phenolchloroform extraction using guanidinisocyanate as a chaotropic agent followed by purification of the lysate on microspin columns. Amplification of fragments of the studied genes was carried out according to the three-step polymerase chain reaction scheme. The number of amplification cycles was 55. The conducted studies showed that the developed complex preparation based on apramycin and cefotaxime has a pronounced antimicrobial activity against potential causative agents of gastrointestinal and respiratory diseases of animals. It was found that the formation of resistance of E. coli 866 strain to cefotaxime and apramycin, as well as to the complex preparation on their basis, is accompanied by significant changes in the functioning of the genetic apparatus of the bacterial cell.

биотрансформация, экспрессия, резистентность, кишечная палочка, ксенобиотики, biotransformation, expression, resistance, Escherichia coli, xenobiotic

1. Гуськова Т.А. Токсикология лекарственных средств / Т.А. Гуськова. -М.: Издательский дом «Русский врач», 2003. -154 с.

2. Коваленко А.М. Иммунопрофилактика и терапия болезней свиней / А.М. Коваленко, Д.А. Евглевский // Белгород-Курск: Курская государственная сельскохозяйственная академия имени профессора И.И. Иванова, 2011. -146 с.

3. Райс Р.Х. Биологические эффекты токсических соединений: курс лекций / Р.Х. Райс, Л.Ф. Гуляева. - Новосибирск: Новосиб. гос. ун-т, 2003. - 208с.

4. Шабунин С.В. Перинатальная патология у крупного рогатого скота - актуальная проблема ветеринарной медицины / С.В.Шабунин, Ю.Н. Алехин, А.Г. Нежданов // Ветеринария. - 2015. - № 1. - С. 3-10.

5. Шабунин С.В. Стратегия и тактика антибактериальной терапии гнойно-воспалительных заболеваний матки у сельскохозяйственных животных / С.В. Шабунин, И.Т. Шапошников, Г.А. Востроилова // Вопросы нормативно-правового регулирования в ветеринарии // 2014. - № 3. - С. 178-181.

6. Шабунин, С.В. Бактериальные и вирусные инфекции в патологии воспроизводительной функции у коров / С.В. Шабунин, А.Г. Шахов, А.Г. Нежданов // Ветеринария. -2012. -№ 10. - С. 3-8.

7. Шахов А.Г. Экологические проблемы здоровья животных и пути их решения/ А.Г. Шахов, М.Н. Аргунов, В.С. Бузлама // Ветеринария. - 2003. - №5. - С. 3-6.

8. Шахов А.Г. Этиология факторных инфекций животных и меры их профилактики/ Шахов А.Г. // Ветеринарная патология. - 2005. - №3. - С. 22-24.

9. Alonso CA. Diversity of Ochrobactrum species in food animals, antibiotic resistance phenotypes and polymorphisms in the blaOCH gene./ Alonso CA, Kwabugge YA, Anyanwu MU, Torres C, Chah KF./ FEMS Microbiol Lett. 2017 Sep 15;364(17). doi: 10.1093/femsle/fnx178. PMID:28911188

10. Chen Z. Effects of chlortetracycline on the fate of multiantibiotic resistance genes and the microbial community during swine manure composting./ Chen Z, Wang Y, Wen Q./ Environ Pollut. 2017 Nov 11. pii: S0269-7491(17)32420-X. doi: 10.1016/ j.envpol.2017.11.009. Epub ahead of print PMID:29137887

11. Livak K.J. Analysis of relative gene expression data using real-time quantitative PCR and the 2-AACt method / K.J. Livak, T.D. Schmittgen // Methods. - 2001. - Vol. 25. - P. 402-408.

12. Kulangara V. Genotypic and phenotypic β-lactam resistance and presence of PVL gene in Staphylococci from dry bovine udder. /Kulangara V, Nair N, Sivasailam A, Sasidharan S, Kollannur JD, Syam R./ PLoS One. 2017 Nov 1;12 (11):e0187277. doi: 10.1371/journal.pone.0187277. eCollection 2017. PMID:29091956

13. Manageiro V. New insights into resistance to colistin and third-generation cephalosporins of Escherichia coli in poultry, Portugal: Novel bla_CTX-M-166 and bla_ESAC genes. / Manageiro V, Clemente L, Graça R, Correia I, Albuquerque T, Ferreira E, Caniça M./ Int J Food Microbiol. 2017 Dec 18;263:67-73. doi: 10.1016/j.ijfoodmicro.2017.10.007. Epub 2017 Oct 4. PMID:29031106

14. Nazir R. Fungal networks serve as novel ecological routes for enrichment and dissemination of antibiotic resistance genes as exhibited by microcosm experiments./ Nazir R, Shen JP, Wang JT, Hu HW, He JZ./ Sci Rep. 2017 Nov 13;7(1): 15457. doi: 10.1038/s41598-017-15660-7. PMID:29133838

15. Wen X. No evidential correlation between veterinary antibiotic degradation ability and resistance genes in microorganisms during the biodegradation of doxycycline./ Wen X, Wang Y, Zou Y, Ma B, Wu Y. / Ecotoxicol Environ Saf. 2018 Jan;147:759-766. doi: 10.1016/j.ecoenv.2017.09.025. Epub 2017 Oct 10. PMID:28942279.