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In Vivo Efficacy of Bacillus velezensis Isolated from Korean Gochang Bokbunja Vinegar against Carbapenem-Resistant Klebsiella pneumoniae Infections Cover

In Vivo Efficacy of Bacillus velezensis Isolated from Korean Gochang Bokbunja Vinegar against Carbapenem-Resistant Klebsiella pneumoniae Infections

Polish Journal of Microbiology
Volume 71 (2022): Issue 4 (December 2022)
By: Fatemeh Ghorbanian,  Hoonhee Seo,  Hanieh Tajdozian,  Youngkyoung Lee,  MD Abdur Rahim,  Sukyung Kim,  Il-Yun Jung,  Saebim Lee and  Ho-Yeon Song  
Open Access
|Dec 2022

Figures & Tables

Fig. 1

Bacterial growth and pH measurement in food grade medium with time. The probiotic strain was inoculated and cultivated. Its A) growth and B) pH were checked every 2 h.
Bacterial growth and pH measurement in food grade medium with time. The probiotic strain was inoculated and cultivated. Its A) growth and B) pH were checked every 2 h.

Fig. 2

Effect of B. velezensis on a clinical isolate of CRKP in the preventive infection murine model. A) Body weight and B) illness severity were observed during the experimental period. Illness severity score was evaluated as (1 - healthy, 2 - minimally ill, 3 - moderately ill, 4 - severely ill, 5 - dead). C) Survival rate was observed during the entire experimental period. D) CRKP treated group (left image), CRKP + B. velezensis treated group (right image). Statistical significance with the control was analyzed using unpaired Student’s f-test (*** p<0.001; **p<0.01; * p<0.05).
Effect of B. velezensis on a clinical isolate of CRKP in the preventive infection murine model. A) Body weight and B) illness severity were observed during the experimental period. Illness severity score was evaluated as (1 - healthy, 2 - minimally ill, 3 - moderately ill, 4 - severely ill, 5 - dead). C) Survival rate was observed during the entire experimental period. D) CRKP treated group (left image), CRKP + B. velezensis treated group (right image). Statistical significance with the control was analyzed using unpaired Student’s f-test (*** p<0.001; **p<0.01; * p<0.05).

Fig. 3

Effect of B. velezensis on a clinical isolate of CRKP in a therapeutic infection murine model. A) Body weight, B) severity of illness, and C) survival rate were observed during the experimental period. Illness severity score was evaluated (1 - healthy, 2 - minimally ill, 3 - moderately ill, 4 - severely ill, 5 - dead). D) CRKP treated group (left image), and CRKP + B. velezensis treated group (right image). Statistical significance with the control was analyzed using unpaired Students f-test (*** p <0.001; ** p< 0.01; * p <0.05).
Effect of B. velezensis on a clinical isolate of CRKP in a therapeutic infection murine model. A) Body weight, B) severity of illness, and C) survival rate were observed during the experimental period. Illness severity score was evaluated (1 - healthy, 2 - minimally ill, 3 - moderately ill, 4 - severely ill, 5 - dead). D) CRKP treated group (left image), and CRKP + B. velezensis treated group (right image). Statistical significance with the control was analyzed using unpaired Students f-test (*** p <0.001; ** p< 0.01; * p <0.05).

Fig. 4

Two-week repeated oral dose toxicity test of B. velezensis using guinea pigs. The body weights of guinea pigs were measured on days 1, 3, 5, 7, 9, 10, and 14. The candidate probiotic was orally administered at 2 x 10s CFU/mice/day at 200 pi per animal once a day, every day, for 2 weeks.
Two-week repeated oral dose toxicity test of B. velezensis using guinea pigs. The body weights of guinea pigs were measured on days 1, 3, 5, 7, 9, 10, and 14. The candidate probiotic was orally administered at 2 x 10s CFU/mice/day at 200 pi per animal once a day, every day, for 2 weeks.

The ability of the candidate probiotic strain to dehydrate carbon sources_

No.Type of testCandidate strainBacillus velezensis KCTC 13417Bacillus amyloliquefaciens KCTC 3002Bacillus licheniformis KCTC 1659
24 h48 h24 h48 h24 h48 h24 h48 h
0Control--------
1Glycerol+++++-++
2Erythritol--------
3D-arabinose--------
4L-arabinose-+++++-+
5D-ribose++++++++
6D-xylose-+++++--
7L-xylose--------
8D-adonite--------
9Methyl-pD-xylopyranoside--------
10D-galactose++++----
11D-glucose+++++-+-
12D-fructose+++++-+-
13D-mannose+++++-+-
14L-sorbose---+----
15L-rhamnose++++----
16Dulcitol--------
17Inocitol++++++--
18D-mannitol+++++-+-
19D-sorbito+++++-++
20Methyl-aD-mannopyranoside--------
21Methyl-aD-glucopyranoside-+++++++
22N-acetylglucosamine++--+-+-
23Amygdaline++++-++-
24Arbutine+++++-+-
25Esculine++++++++
26Salicin+++-+++-
27D-Cellibiose++++++++
28D-Maltose++++++++
29D-Lactose++--++++
30D-Melibiose-+-+++-+
31D-Sacharose+++-+++-
32D-Trehalose++++++--
33Inulin---+----
34D-Melezitose+-------
35D-Raffinose++++++++
36Amidon+++-+-+-
37Glycogen-++-+-+-
38Xylitol--------
39Gentibiose+-----+-
40D-Turanose--++--++
41D-Lyxose--------
42D-Tagatose+++-----
42D-Fucose--------
44L-Fucose--------
45D-arabitol--------
46L-arabitol--------
47Potassium gluconate+-------
48Potassium 2 ketogluconate--------
49Potassium 5 ketogluconate--------

Identification of isolated bacterial strain based on 16S rRNA gene sequence analysis_

NCBI referencesOrganismsLengthScoreIdentitiesGaps
NR_075005.2Bacillus velezensis strain FZB421,5502,728 bits (1,477)1,492/1,499 (99%)2/1,499 (0%)
NR_116240.1Bacillus velezensis strain CBMB2051,4452,615 bits (1,416)1,427/1,432 (99%)2/1,432 (0%)
NR_112685.1Bacillus amyloliquefaciens strain NBRC 155351,4752,699 bits (1,461)1,470/1,475 (99%)0/1,475 (0%)
NR_041455.1Bacillus amyloliquefaciens strain NBRC 155351,4722,697 bits (1,460)1,468/1,472 (99%)0/1,472 (0%)
NR_102783.2Bacillus subtilis subsp. subtilis strain 1681,5502712 bits (1,468)1,489/1,499 (99%)2/1,499 (0%)
NR_118996.1Bacillus licheniformis strain DSM 131,5452,579 bits (1,396)1,466/1,500 (98%)4/1,500 (0%)
NR_116023.1Bacillus licheniformis strain BCRC 117021,4682,545 bits (1,378)1,439/1,469 (98%)2/1,469 (0%)

Identification of the isolated bacterial strain by qRT-PCR_

Species-specific primer for-Oligo nameAmplification results
Candidate probiotic strainB. velezensis KCTC 13417B. atnyloliquefaciens KCTC 3002B. subtilis KCTC 3135B. lichenifortnis KCTC 1659
Bacillus velezensisBvel++---
Bacillus subtilisYtcP---+-
Bacillus atnyloliquefaciensspBamyphes+++--
Bacillus lichenifortnisBlich----+
MacrolactinMin++---
DOI: https://doi.org/10.33073/pjm-2022-051 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 553 - 562
Submitted on: Sep 9, 2022
|
Accepted on: Nov 3, 2022
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Published on: Dec 12, 2022
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
carbapenem-resistant (CRE),
Gochang Bokbunja vinegar,
probiotics
Related subjects:
Life sciences,
Microbiology and virology

© 2022 Fatemeh Ghorbanian, Hoonhee Seo, Hanieh Tajdozian, Youngkyoung Lee, MD Abdur Rahim, Sukyung Kim, Il-Yun Jung, Saebim Lee, Ho-Yeon Song, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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