Russian Journal of Infection and ImmunityRussian Journal of Infection and ImmunityИнфекция и иммунитет2220-76192313-7398SPb RAACI158910.15789/2220-7619-MDR-1589ORIGINAL ARTICLESОРИГИНАЛЬНЫЕ СТАТЬИMulti-drug resistance and biofilm production among diarrheagenic Escherichia coli pathotypes isolated from stools of children with acute diarrheal diseaseМножественная лекарственная устойчивость и образование биопленок среди диарейных патотипов Escherichia coli, выделенных из стула детей с острой диареейhttps://orcid.org/0000-0001-6903-6392DavidE. E.ДэвидЭ. Э.
Nigeria

Ebuka Elijah David,  MSc (Biochemstry), Department of Chemistry/Biochemistry 

1010, Ebonyi State, Abakaliki

Phone: +234 803 318-88-23 

Эбука Элайджа Давид,  магистр биохимии, кафедра химии/биохимии 

1010, штат Эбони, Абакалики

Тел.: +234 803 318-88-23

david.ebuka@funai.edu.ngYameenM. A.ЯминМ. А.
Pakistan

PhD (Microbial Pharmacology), Department of Pharmacy 

Abbottabad

к.н. (фармакология антимикробных препаратов), кафедра аптечной службы 

г. Абботтабад

arfatyameen@gmail.comIgwenyiI.ИгвениИ.
Nigeria

PhD (Biochemistry), Associate Professor, Department of Biochemistry 

Ebonyi State

к.н. (биохимия), доцент кафедры биохимии 

штат Эбони

igweyike@yahoo.comOkaforA. C.ОкафорА. С.
Nigeria

PhD (Microbiology), Department of Microbiology 

Edo State

к.н. (фармакология), кафедра микробиологии 

г. Иямхо, штат Эдо

arthur.okafor@edouniversity.edu.ngObetenU. N.ОбетенУ. Н.
Nigeria

MSc (Biochemstry), Department of Chemistry/Biochemistry 

Ikwo, Ebonyi State

магистр биохимии, кафедра химии/биохимии 

Икво, штат Эбони

uketgabriel@gmail.comObasiD. O.ОбасиД. О.
Nigeria

MSc (Biochemstry), Department of Chemistry/Biochemistry 

Ikwo, Ebonyi State

магистр биохимии, кафедра химии/биохимии 

Икво, штат Эбони

obasidoris19@gmail.comEzeiloU. R.ЭзейлоУ. Р.
Nigeria

PhD (Enzymology), Department of Chemistry/Biochemistry 

Ikwo, Ebonyi State

к.н. (энзимология), кафедра химии/биохимии 

Икво, штат Эбони

ucmgbenka@gmail.comEmeriboleM. N.ЭмериболеМ. Н.
Nigeria

MSc (Biochemstry), Department of Chemistry/Biochemistry 

Ikwo, Ebonyi State

магистр биохимии, кафедра химии/биохимии 

Икво, штат Эбони

mnnanyere@gmail.comDavidC. N.ДэвидЧ. Н.
Nigeria

MSc (Biochemstry), Department of Microbiology 

Ebonyi State

магистр микробиологии, кафедра микробиологии 

штат Эбони

nnenne.nnaji@gmail.comAlex Ekwueme Federal University Ndufu-AlikeФедеральный университет Алекса Эквуэме «Ндуфу Алик Икво»COMSATS University Islamabad, Abbottabad CampusУниверситет COMSATS в ИсламабадеEbonyi State UniversityГосударственный университет ЭбониEdo University, IyamhoУниверситет Эдо211120211159589642208202017042021Copyright ©; 2021, David E.E., Yameen M.A., Igwenyi I., Okafor A.C., Obeten U.N., Obasi D.O., Ezeilo U.R., Emeribole M.N., David C.N.Copyright ©; 2021, Дэвид Э.Э., Ямин М.А., Игвени И., Окафор А.С., Обетен У.Н., Обаси Д.О., Эзейло У.Р., Эмериболе М.Н., Дэвид Ч.Н.2021David E.E., Yameen M.A., Igwenyi I., Okafor A.C., Obeten U.N., Obasi D.O., Ezeilo U.R., Emeribole M.N., David C.N.Дэвид Э.Э., Ямин М.А., Игвени И., Окафор А.С., Обетен У.Н., Обаси Д.О., Эзейло У.Р., Эмериболе М.Н., Дэвид Ч.Н.https://creativecommons.org/licenses/by/4.0https://iimmun.ru/iimm/article/view/1589

Background. Diarrheagenic E. coli (DEC) is an etiological agent of childhood diarrhea. Resistance against commonly used drugs in the empirical treatment of enteric infections has increased among DEC. Relationship between antibiotic resistance and biofilm formation in microorganisms have been widely reported. This study was aimed to determine the antibiotic resistance and biofilm production pattern among DEC pathotypes isolated from stools of children aged 0–5 years with acute diarrheal disease in Abakaliki, Nigeria. Materials and methods. Diarrheal stool samples were obtained from 60 children and E. coli were isolated and identified using standard guidelines provided for laboratory diagnosis of enteric pathogens. Molecular identification was done by amplification of E. coli universal stress protein A (uspA) using polymerase chain reaction (PCR) method. Detection of virulent genes of DEC pathotypes was performed in a group of multiplex PCR using their specific primers. Kirby–Bauer disk diffusion method was used to determine the antibiotic susceptibility patterns of the isolates while biofilms production was detected by thiazolyl blue tetrazolium bromide dye in a 96-well plate. Results. DEC was isolated in 40 stools among which EIEC [40% (n = 16)] was commonly detected followed by ETEC [30% (n = 12)], EAEC [20% (n = 8)] and typical EPEC [10% (n = 4)]. Half of EAEC showed the highest multidrug resistance against ampicillin, cefoxitin, ciprofloxacin, levofloxacin, and tetracycline with the strongest biofilm production followed by all the EPEC which were resistant to ampicillin, ciprofloxacin, levofloxacin, and tetracycline with moderate biofilm production. All the LT-ETEC exhibited the least resistance to ampicillin and tetracycline with the weakest biofilm production. Conclusion. High frequency of the EIEC pathotype suggests its role as the primary etiological agent of diarrhea in children. Correlation between high drug resistance and biofilm production among the pathotype may indicate that biofilms may provide compatible uptake of resistance genes.

Актуальность. Диареагенная кишечная палочка (DEC) является этиологическим агентом диареи у детей. Устойчивость к лекарствам, обычно используемым при эмпирическом лечении кишечных инфекций, среди DEC увеличилась. Широко известна взаимосвязь между устойчивостью к антибиотикам и образованием биопленок у микроорганизмов. Настоящее исследование было направлено на определение устойчивости к анти биотикам и характера биопленкообразования среди патотипов DEC, выделенных из стула детей в возрасте 0–5 лет с острой диареей в Абакалики, Нигерия. Материалы и методы. Образцы стула были получены от 60 детей, и с использованием стандартных подходов, предусмотренных для лабораторной диагностики кишечных патогенов, были выделены и идентифицированы E. coli. Молекулярную идентификацию проводили путем амплификации участка гена универсального стрессового белка А (uspA) E. coli с использованием метода полимеразной цепной реакции (ПЦР). Выявление вирулентных генов патотипов DEC проводили с применением мультиплексной ПЦР со специфическими праймерами. Метод дисковой диффузии Кирби–Бауэра был использован для определения характеристик чувствительности изолятов к антибиотикам, в то время как образование биопленок определяли с помощью красителя тиазолила синего тетразолия бромида в 96-луночном планшете. Результаты. DEC была выделена в 40 образцах. Наиболее часто выявлялась энтероинвазивная E. coli (EIEC) [40% (n = 16)], затем энтеротоксигенная E. coli (ЕТЕС) [30% (n = 12)], энтероагрегативная к E. coli (ЕАЕС) [20% (n = 8)] и типичная энтеропатогенная E. coli (ЕРЕС) [10% (n = 4)]. Половина штаммов EAEC обнаружила самую высокую множественную лекарственную устойчивость к ампициллину, цефокситину, ципрофлоксацину, левофлоксацину и тетрациклину с самым сильным образованием биопленок. Далее по степени устойчивости к антибиотикам и уровню образования биопленок следовали EPEC, которые были устойчивы к ампициллину, ципрофлоксацину, левофлоксацину и тетрациклину и характеризовались умеренным биопленкобразованием. ЕТЕС с термолабильным токсином LT-ETEC проявляли наименьшую устойчивость к ампициллину и тетрациклину с наименьшим образованием биопленок. Заключение. Высокая частота патотипа EIEC свидетельствует о его роли в качестве основного этиологического агента диареи у детей. Корреляция между высокой лекарственной устойчивостью и производством биопленок среди патотипов может указывать на то, что биопленки могут обеспечивать оптимальный уровень захвата генов устойчивости.

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