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

NCBI references	Organisms	Length	Score	Identities	Gaps
NR_075005.2	Bacillus velezensis strain FZB42	1,550	2,728 bits (1,477)	1,492/1,499 (99%)	2/1,499 (0%)
NR_116240.1	Bacillus velezensis strain CBMB205	1,445	2,615 bits (1,416)	1,427/1,432 (99%)	2/1,432 (0%)
NR_112685.1	Bacillus amyloliquefaciens strain NBRC 15535	1,475	2,699 bits (1,461)	1,470/1,475 (99%)	0/1,475 (0%)
NR_041455.1	Bacillus amyloliquefaciens strain NBRC 15535	1,472	2,697 bits (1,460)	1,468/1,472 (99%)	0/1,472 (0%)
NR_102783.2	Bacillus subtilis subsp. subtilis strain 168	1,550	2712 bits (1,468)	1,489/1,499 (99%)	2/1,499 (0%)
NR_118996.1	Bacillus licheniformis strain DSM 13	1,545	2,579 bits (1,396)	1,466/1,500 (98%)	4/1,500 (0%)
NR_116023.1	Bacillus licheniformis strain BCRC 11702	1,468	2,545 bits (1,378)	1,439/1,469 (98%)	2/1,469 (0%)

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.

Table II

The ability of the candidate probiotic strain to dehydrate carbon sources.

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

[i] (+) - positive reaction (yellow) (No. 25: black); (-) - negative reaction (red)

Table III

Identification of the isolated bacterial strain by qRT-PCR.

Species-specific primer for-	Oligo name	Amplification results
Species-specific primer for-	Oligo name	Candidate probiotic strain	B. velezensis KCTC 13417	B. atnyloliquefaciens KCTC 3002	B. subtilis KCTC 3135	B. lichenifortnis KCTC 1659
Bacillus velezensis	Bvel	+	+	-	-	-
Bacillus subtilis	YtcP	-	-	-	+	-
Bacillus atnyloliquefaciens	spBamyphes	+	+	+	-	-
Bacillus lichenifortnis	Blich	-	-	-	-	+
Macrolactin	Min	+	+	-	-	-

[i] (+) - amplification, (-) - no amplification

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 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).

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.

Articles in this issue

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

Published on: Dec 12, 2022

Published by: Polish Society of Microbiologists

In partnership with: Paradigm Publishing Services

Publication frequency: 4 issues per year

Keywords:

Bacillus velezensis,

Klebsiella pneumoniae,

carbapenem-resistant Enterobacteriaceae (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.

Volume 71 (2022): Issue 4 (December 2022)

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

Figures & Tables

Table I

Fig. 1

Table II

Table III

Fig. 2

Fig. 3

Fig. 4

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