Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains

Obligate anaerobic difficult culture bacteria, which make up the bulk of the microbiota of the large intestine, are potential candidates for the development of highly effective new generation probiotics, since they are capable of synthesizing a variety of metabolites, including short-chain fatty aci...

Full description

Saved in:
Bibliographic Details
Main Authors: B O. Bembeeva, E. L. Isaeva, V. V. Muravieva, K. N. Zhigalova, O. V. Nechaeva, D. Kh. Bazukheyr, T. V. Priputnevich
Format: Article
Language:Russian
Published: Numikom LLC 2025-01-01
Series:Эпидемиология и вакцинопрофилактика
Subjects:
Online Access:https://www.epidemvac.ru/jour/article/view/2120
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849411540606779392
author B O. Bembeeva
E. L. Isaeva
V. V. Muravieva
K. N. Zhigalova
O. V. Nechaeva
D. Kh. Bazukheyr
T. V. Priputnevich
author_facet B O. Bembeeva
E. L. Isaeva
V. V. Muravieva
K. N. Zhigalova
O. V. Nechaeva
D. Kh. Bazukheyr
T. V. Priputnevich
author_sort B O. Bembeeva
collection DOAJ
description Obligate anaerobic difficult culture bacteria, which make up the bulk of the microbiota of the large intestine, are potential candidates for the development of highly effective new generation probiotics, since they are capable of synthesizing a variety of metabolites, including short-chain fatty acids, which have a stimulating effect on both commensal bacteria and host cells. However, for their long-term preservation, it is necessary to carefully select the preservation method and protective components. To evaluate the efficacy of using different cryoprotectants to increase the viability of obligate anaerobic difficult culture bacteria when preserved by lyophilisation and cryopreservation methods. The studies showed that freeze-drying contributed most to the preservation of the viability of Faecalibacterium prausnitzii, Anaerostipes hadrus, Eubacterium hallii, provided that they were used as a stabilizing medium, which included inulin, cysteine and riboflavin, ensuring the preservation of the initial bacterial titer for 30 days. Among the studied bacteria, the effectiveness of cryopreservation was shown for A. hadrus, since it ensured the survival of bacteria at the initial level for 14 days of storage, regardless of the cryopreservative used, but by the 30th day their viability decreased significantly (by 100 times when using liquid as a cryopreservant). nutrient medium 110 with minced meat and carbohydrates with the addition of glycerin, 10 000 with the commercial cryopreservative CRYOINSTANT). The results obtained allow us to recommend lyophilization as the most optimal method for long-term storage of probiotic strains of bacteria using highly effective stabilizers.
format Article
id doaj-art-cc7553b058674bb590b99d6cb1436dc7
institution Kabale University
issn 2073-3046
2619-0494
language Russian
publishDate 2025-01-01
publisher Numikom LLC
record_format Article
series Эпидемиология и вакцинопрофилактика
spelling doaj-art-cc7553b058674bb590b99d6cb1436dc72025-08-20T03:34:44ZrusNumikom LLCЭпидемиология и вакцинопрофилактика2073-30462619-04942025-01-01236546010.31631/2073-3046-2024-23-6-54-601082Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic StrainsB O. Bembeeva0E. L. Isaeva1V. V. Muravieva2K. N. Zhigalova3O. V. Nechaeva4D. Kh. Bazukheyr5T. V. Priputnevich6National Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian FederationNational Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian Federation;National Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian FederationNational Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian FederationNational Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian FederationNational Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian FederationNational Medical Research Center for Obstetrics, Gynecology and Perinatology named after Academician V.I. Kulakov of Ministry of Health of Russian Federation; Federal State Autonomous Educational Institution of Higher Education «N.I. Pirogov Russian National Research Medical University» of the Ministry of Health of the Russian Federation; Federal State Budgetary Educational Institution of Further Professional Education "Russian Medical Academy of Continuous Professional Education" of the Ministry of Healthcare of the Russian FederationObligate anaerobic difficult culture bacteria, which make up the bulk of the microbiota of the large intestine, are potential candidates for the development of highly effective new generation probiotics, since they are capable of synthesizing a variety of metabolites, including short-chain fatty acids, which have a stimulating effect on both commensal bacteria and host cells. However, for their long-term preservation, it is necessary to carefully select the preservation method and protective components. To evaluate the efficacy of using different cryoprotectants to increase the viability of obligate anaerobic difficult culture bacteria when preserved by lyophilisation and cryopreservation methods. The studies showed that freeze-drying contributed most to the preservation of the viability of Faecalibacterium prausnitzii, Anaerostipes hadrus, Eubacterium hallii, provided that they were used as a stabilizing medium, which included inulin, cysteine and riboflavin, ensuring the preservation of the initial bacterial titer for 30 days. Among the studied bacteria, the effectiveness of cryopreservation was shown for A. hadrus, since it ensured the survival of bacteria at the initial level for 14 days of storage, regardless of the cryopreservative used, but by the 30th day their viability decreased significantly (by 100 times when using liquid as a cryopreservant). nutrient medium 110 with minced meat and carbohydrates with the addition of glycerin, 10 000 with the commercial cryopreservative CRYOINSTANT). The results obtained allow us to recommend lyophilization as the most optimal method for long-term storage of probiotic strains of bacteria using highly effective stabilizers.https://www.epidemvac.ru/jour/article/view/2120difficult culture bacteriaobligate anaerobeslyophilisationcryopreservationprobiotics no conflict of interest to declare
spellingShingle B O. Bembeeva
E. L. Isaeva
V. V. Muravieva
K. N. Zhigalova
O. V. Nechaeva
D. Kh. Bazukheyr
T. V. Priputnevich
Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
Эпидемиология и вакцинопрофилактика
difficult culture bacteria
obligate anaerobes
lyophilisation
cryopreservation
probiotics no conflict of interest to declare
title Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
title_full Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
title_fullStr Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
title_full_unstemmed Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
title_short Optimising Conditions for the Preservation of Extremely Oxygen Sensitive Obligate Anaerobes of the Gut Microbiota as Candidates for Probiotic Strains
title_sort optimising conditions for the preservation of extremely oxygen sensitive obligate anaerobes of the gut microbiota as candidates for probiotic strains
topic difficult culture bacteria
obligate anaerobes
lyophilisation
cryopreservation
probiotics no conflict of interest to declare
url https://www.epidemvac.ru/jour/article/view/2120
work_keys_str_mv AT bobembeeva optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT elisaeva optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT vvmuravieva optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT knzhigalova optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT ovnechaeva optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT dkhbazukheyr optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains
AT tvpriputnevich optimisingconditionsforthepreservationofextremelyoxygensensitiveobligateanaerobesofthegutmicrobiotaascandidatesforprobioticstrains