scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agencyscbmt-105Research ArticleМЕТОДЫ БИОМЕДИЦИНСКИХ ИССЛЕДОВАНИЙMETHODS OF BIOMEDICAL RESEARCHESАнтагонистическая активность консорциума штаммов Lactobacillusfermentum ВКМ В-2793D и Lactobacillusreuteri ВКМ В-3144D, выделенных из женского молока, к мастит-индуцирующим патогенамAntagonistic activity of a consortium of strains Lactobacillus fermentum ВКМ В-2793D and Lactobacillus reuteri ВКМ В-3144D isolated from human milk to mastitis-inductioning pathogensАбрамовВ. М.AbramovV. M.noemail@neicon.ruХлебниковВ. С.KhlebnikovV. S.noemail@neicon.ruКосаревИ. В.KosarevI. V.kosarev-52@mail.ruПрипутневичТ. В.PriputnevichT. V.noemail@neicon.ruЛюбасовскаяЛ. А.LyubasovskayaL. A.noemail@neicon.ruВасиленкоР. Н.VasilenkoR. N.noemail@neicon.ruСакулинВ. К.SakulinV. K.noemail@neicon.ruКаркищенкоВ. Н.KarkischenkoV. N.noemail@neicon.ruКаркищенкоН. Н.KarkischenkoN. N.noemail@neicon.ruПчелинцевС. Ю.PchelintsevS. Yu.noemail@neicon.ruКарлышевА. В.KarlyshevA. V.noemail@neicon.ruСамойленкоВ. А.SamojlenkoV. A.noemail@neicon.ruМачулинА. В.MachulinA. V.noemail@neicon.ruОАО «Институт инженерной иммунологии»РоссияФГБУ «Национальный медицинский исследовательский центр акушерства, гинекологии и перинатологии имени акад. В.И. Кулакова» Минздрава РоссииРоссияФГБУН «Научный центр биомедицинских технологий ФМБА России»РоссияКингстонский университетРоссияФГБУН «Институт биохимии и физиологии микроорганизмов им. Г.К. Скрябина» РАНРоссия201826022019034654Copyright © Абрамов В.М., Хлебников В.С., Косарев И.В., Припутневич Т.В., Любасовская Л.А., Василенко Р.Н., Сакулин В.К., Каркищенко В.Н., Каркищенко Н.Н., Пчелинцев С.Ю., Карлышев А.В., Самойленко В.А., Мачулин А.В., 20192019Абрамов В.М., Хлебников В.С., Косарев И.В., Припутневич Т.В., Любасовская Л.А., Василенко Р.Н., Сакулин В.К., Каркищенко В.Н., Каркищенко Н.Н., Пчелинцев С.Ю., Карлышев А.В., Самойленко В.А., Мачулин А.В.Abramov V.M., Khlebnikov V.S., Kosarev I.V., Priputnevich T.V., Lyubasovskaya L.A., Vasilenko R.N., Sakulin V.K., Karkischenko V.N., Karkischenko N.N., Pchelintsev S.Y., Karlyshev A.V., Samojlenko V.A., Machulin A.V.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/105

Целью работы явилось изучение антагонистической активности штаммов Lactobacillus fermentum BKM B-2793D, Lactobacillus reuteri BKM B-3144D, выделенных из женского молока, а также их консорциума к патогенам, вызывающим мастит в лактационный период. Установлено, что лактобактерии и их консорциум проявляют антагонистическую активность к индикаторным культурам, использованным в работе, включая резистентные к антибиотикам патогены. Консорциум обладает более выраженной антагонистической активностью к мастит-индуцирующим патогенам по сравнению с отдельными штаммами лактобактерий, входящими в его состав. Консорциум защищает клетки MCF-7 молочной железы человека от адгезии и колонизации патогеном S.aureus 8325-4. Полученные результаты свидетельствуют о том, что лактобактерии L.fermentum BKM B-2793D, L.reuteri BKM B-3144D, а также их консорциум являются отличными кандидатами для разработки пробиотического препарата для профилактики и лечения лактационного мастита.

The aim of the study was to study the antagonistic activity of strains of Lactobacillus fermentum BKM B-2793D, Lactobacillus reuteri BKM B-3144D isolated from human breast milk and their consortium to pathogens that cause mastitis during the lactation period. The lactobacilli and their consortium reveal antimicrobial activity against the indicator microorganisms used in this study including pathogens resistant to antibiotics. The consortium protects human mammary gland MCF-7 cells against adhesion and colonization by Staphylococcus aureus 8325-4. The obtained results suggest that lactobacilli from breast milk of healthy woman L. fermentum ВКМ В-2793D and L. reuteri ВКМ В-3144D and their consortium are excellent candidates for the development of probiotic preparation for prophylactics and treatment of lactational mastitis.

лактобактерии женского молокаантагонистическая активностьбиомоделированиеlactobacilli of human milkantagonistic activitybiomodellingReferencesРуководство по проведению доклинических исследований лекарственных средств (иммунобиологические лекарственные препараты). Ч. 2 / Под ред. А.Н. Миронова. - М.: Гриф и К. - 2012. - 536 с.Руководство по проведению доклинических исследований лекарственных средств (иммунобиологические лекарственные препараты). Ч. 2 / Под ред. А.Н. Миронова. - М.: Гриф и К. - 2012. - 536 с.Brouillette E., Talbot B., Malouin F. The fibronectin-binding proteins of Staphulococcus aureus may promote mammary gland colonization in a lactating mouse model of Mastitis // Infect. Immun. 2003. V. 71. No. 4. Pp. 2292-2295.Brouillette E., Talbot B., Malouin F. The fibronectin-binding proteins of Staphulococcus aureus may promote mammary gland colonization in a lactating mouse model of Mastitis // Infect. Immun. 2003. V. 71. No. 4. Pp. 2292-2295.Brouillette E., Martinez A., Boyll B., et al. Persistence of a Staphylococcus aureus Small-colony variant under antibiotic pressure in vivo // FEMS Immunol. and med. microbiol. 2004. V. 41. Issue 1. Pp. 35-41.Brouillette E., Martinez A., Boyll B., et al. Persistence of a Staphylococcus aureus Small-colony variant under antibiotic pressure in vivo // FEMS Immunol. and med. microbiol. 2004. V. 41. Issue 1. Pp. 35-41.Delgado S., Arroyo R., Jiménez E., et al. Staphylococcus epidermidis strains isolated from breast milk of women suffering infectious mastitis: potential virulence traits and resistance to antibiotics // BMC Microbiol. 2009. V. 9. Pp. 82-85.Delgado S., Arroyo R., Jiménez E., et al. Staphylococcus epidermidis strains isolated from breast milk of women suffering infectious mastitis: potential virulence traits and resistance to antibiotics // BMC Microbiol. 2009. V. 9. Pp. 82-85.Delgado S., Arroyo R., Jiménez E., et al. Infectious mastitis during lactation: an underrated condition (I) // Acta Pediatrica Espanola. 2009. V. 67. Pp. 77-84.Delgado S., Arroyo R., Jiménez E., et al. Infectious mastitis during lactation: an underrated condition (I) // Acta Pediatrica Espanola. 2009. V. 67. Pp. 77-84.Delgado S., Garcia P., Fernandez L., Jimenez E., Rodriguez-Banos M., Del Campo R., Rodriguez J.M. Characterization of Staphylococcus aureus strains involved in human and bovine mastitis // FEMS immunological and medical microbiology. 2011. V. 62. Pp. 225-235.Delgado S., Garcia P., Fernandez L., Jimenez E., Rodriguez-Banos M., Del Campo R., Rodriguez J.M. Characterization of Staphylococcus aureus strains involved in human and bovine mastitis // FEMS immunological and medical microbiology. 2011. V. 62. Pp. 225-235.Fernández L., Arroyo R., Espinosa I., Marín M, Jiménez E., Rodríguez J.M. Probiotics for human lactational mastitis // Benef. Microb. 2014. V. 5. Pp. 169-183.Fernández L., Arroyo R., Espinosa I., Marín M, Jiménez E., Rodríguez J.M. Probiotics for human lactational mastitis // Benef. Microb. 2014. V. 5. Pp. 169-183.Indrio F., Neu J. The intestinal microbiome of infants and the use of probiotics // Cur. Opin. Pediatr. 2011. V. 23. No. 2. Pp. 145-150.Indrio F., Neu J. The intestinal microbiome of infants and the use of probiotics // Cur. Opin. Pediatr. 2011. V. 23. No. 2. Pp. 145-150.Jost T., Lacroix C., Braegger C., Rochat F., Chassart C. Vertical mother-neonate transfer of maternal gut bacteria via breast-feeding // Environmental Microbiology. 2013. doi:10.1111/1462-2920.12238.Jost T., Lacroix C., Braegger C., Rochat F., Chassart C. Vertical mother-neonate transfer of maternal gut bacteria via breast-feeding // Environmental Microbiology. 2013. doi:10.1111/1462-2920.12238.Lowy F.D. Staphulococcus aureus infections // N. Engl. J. Med. 1998. V. 339. Pp. 520-532.Lowy F.D. Staphulococcus aureus infections // N. Engl. J. Med. 1998. V. 339. Pp. 520-532.Lehri B., Seddon A., Karlyshev A. Potential probiotic-associated traits revealed from completed high quality genome sequence of Lacobacillus fermentum 3872 // Standards in Genomic Sciences. 2017. doi:10.1186/s40793-017-0228-4.Lehri B., Seddon A., Karlyshev A. Potential probiotic-associated traits revealed from completed high quality genome sequence of Lacobacillus fermentum 3872 // Standards in Genomic Sciences. 2017. doi:10.1186/s40793-017-0228-4.Lehri B., Seddon A., Karlyshev A. Lactobacillus fermentum 3872 genome sequencing reveals plasmid and chromosomal genes potentially involved in a probiotic activity // FEMS Microbiol. Let. 2015. V. 362. Pp. 1-5.Lehri B., Seddon A., Karlyshev A. Lactobacillus fermentum 3872 genome sequencing reveals plasmid and chromosomal genes potentially involved in a probiotic activity // FEMS Microbiol. Let. 2015. V. 362. Pp. 1-5.Martin V., Maldonado A., Moles L., et al. Sharing of bacterial strains between breast milk and infant feces // J. of Human Lactation. 2012. V. 28. Pp. 36-44.Martin V., Maldonado A., Moles L., et al. Sharing of bacterial strains between breast milk and infant feces // J. of Human Lactation. 2012. V. 28. Pp. 36-44.Ramiah K., van Reenen C., Dicks L. Surface-bound proteins of Lactobacillus plantarum 423 that contribute to adhesion of Caco-2 cells and their role in competitive exclusion and displacement of Clostridium sporogenes and Enterococcus faecalis // Res. Microbiol. 2008. V. 6. Pp. 470-475.Ramiah K., van Reenen C., Dicks L. Surface-bound proteins of Lactobacillus plantarum 423 that contribute to adhesion of Caco-2 cells and their role in competitive exclusion and displacement of Clostridium sporogenes and Enterococcus faecalis // Res. Microbiol. 2008. V. 6. Pp. 470-475.Saiman L., Okeefe M., Graham P., Wu F., et al. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women // Clin. Infect. Dis. 2003. V. 37. Pp. 1313-19.Saiman L., Okeefe M., Graham P., Wu F., et al. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women // Clin. Infect. Dis. 2003. V. 37. Pp. 1313-19.Schoenfelder S.M., Lange С., Eckart M., Hennig S., Kozytska S., Ziebuhr W. Successthrough diversity - how Staphylococcus epidermidis establishes as a nosocomial pathogen // Int. J. of Medical Microbiology. 2010. V. 300. Pp. 380-386.Schoenfelder S.M., Lange С., Eckart M., Hennig S., Kozytska S., Ziebuhr W. Successthrough diversity - how Staphylococcus epidermidis establishes as a nosocomial pathogen // Int. J. of Medical Microbiology. 2010. V. 300. Pp. 380-386.Sears P.M., McCarthy K.K. Diagnosis of mastitis for therapy decisions // Veterinary Clinics of North America: Food Animal Practice. 2003. V. 19. Pp. 93-108.Sears P.M., McCarthy K.K. Diagnosis of mastitis for therapy decisions // Veterinary Clinics of North America: Food Animal Practice. 2003. V. 19. Pp. 93-108.Soto A., Martín V., Jiménez E., Mader I., Rodríguez J.M., Fernández L. Lactobacilli and bifidobacteria in human breast milk: influence of antibiotherapy and other host and clinical factors // J. Pediatr. Gastroenterol. Nutr. 2014. V. 59. Pp. 78-88.Soto A., Martín V., Jiménez E., Mader I., Rodríguez J.M., Fernández L. Lactobacilli and bifidobacteria in human breast milk: influence of antibiotherapy and other host and clinical factors // J. Pediatr. Gastroenterol. Nutr. 2014. V. 59. Pp. 78-88.Westerlund B., Korhonen T. Bacterial proteins binding to the mammalian extracellular matrics // Mol. Microbiol. 1993. V. 9. Pp. 687-694.Westerlund B., Korhonen T. Bacterial proteins binding to the mammalian extracellular matrics // Mol. Microbiol. 1993. V. 9. Pp. 687-694.World Health Organization. Mastitis: causes and management. - Geneva: WHO, 2000.World Health Organization. Mastitis: causes and management. - Geneva: WHO, 2000.Rukovodstvo po provedeniyu doklinicheskih issledovanij lekarstvennyh sredstv (immunobiologicheskie lekarstvennye preparaty). Ch. 2 [Guidelines for conducting preclinical studies of drugs (immunobiological drugs). Part 2]. Ed. by A.N. Mironov. Moscow: Grif i K. 2012. 536 p. (In Russian).Rukovodstvo po provedeniyu doklinicheskih issledovanij lekarstvennyh sredstv (immunobiologicheskie lekarstvennye preparaty). Ch. 2 [Guidelines for conducting preclinical studies of drugs (immunobiological drugs). Part 2]. Ed. by A.N. Mironov. Moscow: Grif i K. 2012. 536 p. (In Russian).Brouillette E., Talbot B., Malouin F. The fibronectin-binding proteins of Staphulococcus aureus may promote mammary gland colonization in a lactating mouse model of Mastitis. Infect. Immun. 2003. V. 71. No. 4. Pp. 2292-2295.Brouillette E., Talbot B., Malouin F. The fibronectin-binding proteins of Staphulococcus aureus may promote mammary gland colonization in a lactating mouse model of Mastitis. Infect. Immun. 2003. V. 71. No. 4. Pp. 2292-2295.Brouillette E., Martinez A., Boyll B., et al. Persistence of a Staphylococcus aureus Small-colony variant under antibiotic pressure in vivo. FEMS Immunol. and med. microbiol. 2004. V. 41. Issue 1. Pp. 35-41.Brouillette E., Martinez A., Boyll B., et al. Persistence of a Staphylococcus aureus Small-colony variant under antibiotic pressure in vivo. FEMS Immunol. and med. microbiol. 2004. V. 41. Issue 1. Pp. 35-41.Delgado S., Arroyo R., Jiménez E., et al. Staphylococcus epidermidis strains isolated from breast milk of women suffering infectious mastitis: potential virulence traits and resistance to antibiotics. BMC Microbiol. 2009. V. 9. Pp. 82-85.Delgado S., Arroyo R., Jiménez E., et al. Staphylococcus epidermidis strains isolated from breast milk of women suffering infectious mastitis: potential virulence traits and resistance to antibiotics. BMC Microbiol. 2009. V. 9. Pp. 82-85.Delgado S., Arroyo R., Jiménez E., et al. Infectious mastitis during lactation: an underrated condition (I). Acta Pediatrica Espanola. 2009. V. 67. Pp. 77-84.Delgado S., Arroyo R., Jiménez E., et al. Infectious mastitis during lactation: an underrated condition (I). Acta Pediatrica Espanola. 2009. V. 67. Pp. 77-84.Delgado S., Garcia P., Fernandez L., Jimenez E., Rodriguez-Banos M., Del Campo R., Rodriguez J.M. Characterization of Staphylococcus aureus strains involved in human and bovine mastitis. FEMS immunological and medical microbiology. 2011. V. 62. Pp. 225-235.Delgado S., Garcia P., Fernandez L., Jimenez E., Rodriguez-Banos M., Del Campo R., Rodriguez J.M. Characterization of Staphylococcus aureus strains involved in human and bovine mastitis. FEMS immunological and medical microbiology. 2011. V. 62. Pp. 225-235.Fernández L., Arroyo R., Espinosa I., Marín M, Jiménez E., Rodríguez J.M. Probiotics for human lactational mastitis. Benef. Microb. 2014. V. 5. Pp. 169-183.Fernández L., Arroyo R., Espinosa I., Marín M, Jiménez E., Rodríguez J.M. Probiotics for human lactational mastitis. Benef. Microb. 2014. V. 5. Pp. 169-183.Indrio F., Neu J. The intestinal microbiome of infants and the use of probiotics. Cur. Opin. Pediatr. 2011. V. 23. No. 2. Pp. 145-150.Indrio F., Neu J. The intestinal microbiome of infants and the use of probiotics. Cur. Opin. Pediatr. 2011. V. 23. No. 2. Pp. 145-150.Jost T., Lacroix C., Braegger C., Rochat F., Chassart C. Vertical mother-neonate transfer of maternal gut bacteria via breast-feeding. Environmental Microbiology. 2013. doi:10.1111/1462-2920.12238.Jost T., Lacroix C., Braegger C., Rochat F., Chassart C. Vertical mother-neonate transfer of maternal gut bacteria via breast-feeding. Environmental Microbiology. 2013. doi:10.1111/1462-2920.12238.Lowy F.D. Staphulococcus aureus infections. N. Engl. J. Med. 1998. V. 339. Pp. 520-532.Lowy F.D. Staphulococcus aureus infections. N. Engl. J. Med. 1998. V. 339. Pp. 520-532.Lehri B., Seddon A., Karlyshev A. Potential probiotic-associated traits revealed from completed high quality genome sequence of Lacobacillus fermentum 3872. Standards in Genomic Sciences. 2017. doi:10.1186/s40793-017-0228-4.Lehri B., Seddon A., Karlyshev A. Potential probiotic-associated traits revealed from completed high quality genome sequence of Lacobacillus fermentum 3872. Standards in Genomic Sciences. 2017. doi:10.1186/s40793-017-0228-4.Lehri B., Seddon A., Karlyshev A. Lactobacillus fermentum 3872 genome sequencing reveals plasmid and chromosomal genes potentially involved in a probiotic activity. FEMS Microbiol. Let. 2015. V. 362. Pp. 1-5.Lehri B., Seddon A., Karlyshev A. Lactobacillus fermentum 3872 genome sequencing reveals plasmid and chromosomal genes potentially involved in a probiotic activity. FEMS Microbiol. Let. 2015. V. 362. Pp. 1-5.Martin V., Maldonado A., Moles L., et al. Sharing of bacterial strains between breast milk and infant feces. J. of Human Lactation. 2012. V. 28. Pp. 36-44.Martin V., Maldonado A., Moles L., et al. Sharing of bacterial strains between breast milk and infant feces. J. of Human Lactation. 2012. V. 28. Pp. 36-44.Ramiah K., van Reenen C., Dicks L. Surface-bound proteins of Lactobacillus plantarum 423 that contribute to adhesion of Caco-2 cells and their role in competitive exclusion and displacement of Clostridium sporogenes and Enterococcus faecalis. Res. Microbiol. 2008. V. 6. Pp. 470-475.Ramiah K., van Reenen C., Dicks L. Surface-bound proteins of Lactobacillus plantarum 423 that contribute to adhesion of Caco-2 cells and their role in competitive exclusion and displacement of Clostridium sporogenes and Enterococcus faecalis. Res. Microbiol. 2008. V. 6. Pp. 470-475.Saiman L., Okeefe M., Graham P., Wu F., et al. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women. Clin. Infect. Dis. 2003. V. 37. Pp. 1313-19.Saiman L., Okeefe M., Graham P., Wu F., et al. Hospital transmission of community-acquired methicillin-resistant Staphylococcus aureus among postpartum women. Clin. Infect. Dis. 2003. V. 37. Pp. 1313-19.Schoenfelder S.M., Lange С., Eckart M., Hennig S., Kozytska S., Ziebuhr W. Successthrough diversity - how Staphylococcus epidermidis establishes as a nosocomial pathogen. Int. J. of Medical Microbiology. 2010. V. 300. Pp. 380-386.Schoenfelder S.M., Lange С., Eckart M., Hennig S., Kozytska S., Ziebuhr W. Successthrough diversity - how Staphylococcus epidermidis establishes as a nosocomial pathogen. Int. J. of Medical Microbiology. 2010. V. 300. Pp. 380-386.Sears P.M., McCarthy K.K. Diagnosis of mastitis for therapy decisions. Veterinary Clinics of North America: Food Animal Practice. 2003. V. 19. Pp. 93-108.Sears P.M., McCarthy K.K. Diagnosis of mastitis for therapy decisions. Veterinary Clinics of North America: Food Animal Practice. 2003. V. 19. Pp. 93-108.Soto A., Martín V., Jiménez E., Mader I., Rodríguez J.M., Fernández L. Lactobacilli and bifidobacteria in human breast milk: influence of antibiotherapy and other host and clinical factors. J. Pediatr. Gastroenterol. Nutr. 2014. V. 59. Pp. 78-88.Soto A., Martín V., Jiménez E., Mader I., Rodríguez J.M., Fernández L. Lactobacilli and bifidobacteria in human breast milk: influence of antibiotherapy and other host and clinical factors. J. Pediatr. Gastroenterol. Nutr. 2014. V. 59. Pp. 78-88.Westerlund B., Korhonen T. Bacterial proteins binding to the mammalian extracellular matrics. Mol. Microbiol. 1993. V. 9. Pp. 687-694.Westerlund B., Korhonen T. Bacterial proteins binding to the mammalian extracellular matrics. Mol. Microbiol. 1993. V. 9. Pp. 687-694.World Health Organization. Mastitis: causes and management. Geneva: WHO, 2000.World Health Organization. Mastitis: causes and management. Geneva: WHO, 2000.

The authors declare that there are no conflicts of interest present.