B subtilis

Manufactured by Merck Group

Sourced in United States

B. subtilis is a bacterium commonly used as a laboratory organism for various research and testing purposes. It is a gram-positive, spore-forming bacterium that is known for its ability to thrive in a variety of environments. B. subtilis is frequently employed as a model organism in microbiology, genetics, and biotechnology studies due to its well-understood biology and ease of cultivation.

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Lab products found in correlation

E coli dh5α, by Merck Group (1 mentions) C albicans, by Microbiologics (1 mentions) E faecalis, by Merck Group (1 mentions) K pneumoniae, by Microbiologics (1 mentions) A brasiliensis, by Microbiologics (1 mentions) K pneumoniae, by Merck Group (1 mentions) P aeruginosa, by Microbiologics (1 mentions) S aureus, by Merck Group (1 mentions) P aeruginosa, by Merck Group (1 mentions) S aureus, by American Type Culture Collection (1 mentions) E faecalis, by American Type Culture Collection (1 mentions) Nanoject 2 apparatus, by Drummond (1 mentions) Lps e coli serotype 0111 b4, by Merck Group (1 mentions) Avilamycin, by Merck Group (1 mentions)

8 protocols using b subtilis

Dietary Modulation of Poultry Gut Health

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Chickens were fed a commercial diet as a base (Table-1) without adding antibiotic, which was added in some antimicrobial treatments (AGPP; avilamycin, Elanco, Medellín, Colombia). B. subtilis (Alterion^® Global Company, São Paulo^, Brazil) was used according to the manufacturer’s recommendation, 50 g/ton of feed to guarantee a dose of 10⁸ colony-forming units (CFU) and LPS from E. coli (LPS E. coli, serotype 0111:B4; Sigma-Aldrich, St. Louis, MO, USA) to induce intestinal inflammation in vivo [15 ], which are supplied directly in the feed as follows:

Basal diet 1 (D1): Commercial diet without antimicrobial addition, LPS, and B. subtilis

Diet 2 (D2): D1 plus antimicrobial AGP (avilamycin, 10 ppm)

Diet 3 (D3): D1 plus B. subtilis (50 ppm) to guarantee a daily consumption of 10⁸ CFU/animal and have adequate viability

Diet 4 (D4): D1 plus 1.0 μg LPS/g feed (1 ppm)

Diet 5 (D5): D1 plus 1.0 μg LPS/g feed (1 ppm) and antimicrobial AGP (avilamycin, 10 ppm)

Diet 6 (D6): D1 plus 1.0 μg LPS/g feed (1 ppm) and B. subtilis at a rate of 50 ppm feed

No information on production parameters, the sacrifice of chickens, sample collection procedures, days on which chickens were sacrificed, etc.

Rodríguez S.P., Herrera A.L, & Parra J.E. (2023). Gene expression of pro-inflammatory (IL-8, IL-18, TNF-α, and IFN-γ) and anti-inflammatory (IL-10) cytokines in the duodenum of broiler chickens exposed to lipopolysaccharides from Escherichia coli and Bacillus subtilis. Veterinary World, 16(3), 564-570.

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Isolation and Characterization of Lactobacilli

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All the lactobacilli used in this study were originally isolated from vaginal swabs of healthy women using culture-based methods (27 ). The present study was approved by the Ethical Committee of the Second People's Hospital of Yunnan Province. All volunteers provided written consent and were informed of the purposes of the study. Six strains were identified as L. crispatus, L. gasseri, L. jensenii, L. fermentum, L. delbrueckii and L. johnsonii (detailed in the Results and Table S1). All strains were stored at −80°C in MRS media containing 30% glycerol (v/v) and plated on agarose containing MRS media (Hopebio, Qingdao, China) at 37°C for 96 h before individual colonies were used for routine cultures in liquid MRS medium (27 ).
For the indicated experiments, bacterial cells were exposed to ⁶⁰Co radiation (10 KGy) for 10 h, and inactivation was assessed by inoculating MRS plates. E. coli JM109, E. coli DH5α, and B. subtilis purchased from Sigma-Aldrich (Gallen, Switzerland) were used as controls.

Song J., Lang F., Zhao N., Guo Y, & Zhang H. (2018). Vaginal Lactobacilli Induce Differentiation of Monocytic Precursors Toward Langerhans-like Cells: in Vitro Evidence. Frontiers in Immunology, 9, 2437.

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Standardized Microbial Inoculum Preparation

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S. aureus (American Type Culture Collection, ATCC 6538), E. faecalis (ATCC 29212), B. subtilis spizizenii (ATCC 6633), E. cloacae (ATCC 35030), K. pneumoniae (ATCC 10031), A. brasiliensis (ATCC 16404), P. aeruginosa (ATCC 9027) and C. albicans (ATCC 10231) were obtained from Microbiologics (Minnesota, USA). F. solani (Deutsche Sammlung von Mikroorganismen, DSM 1164) was supplied by Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Germany.
Inocula of S. aureus, E. faecalis, B. subtilis, E. cloacae, A. brasiliensis, P. aeruginosa, C. albicans, K. pneumoniae and F. solani were prepared from lyophilised pellets according to the manufacturer’s protocols or from fresh culture according to McFarland standards to obtain 1.0×10⁷ colony forming units (CFU/mL in physiological solution (0.9% (w/v) NaCl); Merck-Sigma-Aldrich, Darmstadt, Germany.
Tryptone soya broth (20 mL; Bioculturalab, Italy) was used as growth control for C. albicans, A. brasiliensis and F. solani, and thioglycolate medium (20 mL; Bioculturalab) was used as growth control for S. aureus, B. subtilis, P. aeruginosa and K. pneumoniae. Nutrient broth (Nutrient Broth, 20 mL; BioLife, Italy) medium was used as growth control for E. faecalis and E. cloacae.

Giurgola L., Gatto C., Honisch C., Rossi O., Ragazzi E, & D'Amato Tothova J. (2021). Killing efficacy of a new hypothermic corneal storage medium against the micro-organisms frequently found in human donor cornea intended for transplantation. BMJ Open Ophthalmology, 6(1), e000833.

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BpiP Binding to Peptidoglycan Cross-Linking Assay

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Recombinant BpiP (100 μg/ml) was incubated with PG from Bb, E. coli (InvivoGen) or B. subtilis (Sigma) (200 μM muramic acid units of peptidoglycan) in Buffer A (40mM sodium phosphate [pH 6.5] with 0.5% Tween 20) on ice for 2 hrs [85 (link), 86 (link), 89 ]. An equal volume of Buffer B (50mM sodium phosphate [pH8.5] containing DTSSP (10mM) on ice for 30 mins was added followed by centrifugation at 21, 000 x g to pull down the cross-linked complexes. The samples were mixed with 2X SDS-PAGE sample buffer with 10% β-mercaptoethanol (β-ME) and boiled at 95°C for 5 minutes and the supernatant were analyzed on SDS-12% PAGE gel and transferred to PVDF membranes. The membranes were developed with mouse antiserum directed at C-terminal region of BpiP, goat anti-mouse IgG conjugated to HRPO and developed using ECL. BpiP-PG crosslinked complexes are unable to enter the gel but the cleavage of the cross-linker by β-mercaptoethanol treatment prior to electrophoresis facilitates analysis of the interactions of wild type BpiP or BpiP with site-specific changes with borrelial or heterologous PG in the supernatant as described in the flow chart in S3 Fig [86 (link), 87 (link), 89 ].

Chen Y., Vargas S.M., Smith TC I.I., Karna S.L., MacMackin Ingle T., Wozniak K.L., Wormley FL J.r, & Seshu J. (2021). Borrelia peptidoglycan interacting Protein (BpiP) contributes to the fitness of Borrelia burgdorferi against host-derived factors and influences virulence in mouse models of Lyme disease. PLoS Pathogens, 17(4), e1009535.

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Fungal Pathogen Infection in Flies

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For infection by septic injury, flies were injured with a thin tungsten needle previously dipped in a microorganism suspension diluted in PBS at the indicated concentrations. Flies were challenged by natural infection with B. bassiana. 18.4 nL of a solution of B. subtilis (P5985; Sigma-Aldrich) or A. oryzae (P6110; Sigma-Aldrich) proteases, diluted at (1:2000) in PBS or 9.2 nL of a sonicated suspension of M. luteus peptidoglycans (5mg/ml; Sigma-Aldrich) were injected into the fly body cavity (Nanoject II apparatus; Drummond Scientific). When needed, 18.4 nL of E-64 (Sigma) diluted in PBS at 0.5 or 2 mM were injected 2 hours before immune challenge.

Issa N., Guillaumot N., Lauret E., Matt N., Schaeffer-Reiss C., Van Dorsselaer A., Reichhart J.M, & Veillard F. (2018). The Circulating Protease Persephone Is an Immune Sensor for Microbial Proteolytic Activities Upstream of the Drosophila Toll Pathway. Molecular Cell, 69(4), 539-550.e6.

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Isolation and preservation of microorganisms

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Strain FJAT-46737 was isolated from a soil sample from the Huanggang Mountain area, Fujian Province, China, and preserved at the China General Microbiological Culture Collection Center (CGMCC No. 14661). The pathogenic strains R. solanacearum FJAT-91 (CGMCC No. 10692, tomato pathogen), R. solanacearum FJAT-77 (peanut pathogen), Escherichia coli FJAT-301, F. oxysporum f. sp. capsicum FJAT-831, F. oxysporum f. sp. niveum FJAT-30265 and F. oxysporum f. sp. melonis FJAT-9230 were preserved at the Agricultural Bioresources Research Institute, Fujian Academy of Agricultural Sciences, Fujian, China. All the culture media used for bacterial and fungal growth were purchased from AoBoXing Biological Technology Co., Ltd. (Beijing, China). The cultivation substrates used for the pot experiments were purchased from Xiamen Jiang Ping biological Technology Co., Ltd. (Xiamen, China). The reference standards for iturin and surfactin were produced from B. subtilis and purchased from Sigma (St. Louis, MO, USA).

Chen M., Wang J., Liu B., Zhu Y., Xiao R., Yang W., Ge C, & Chen Z. (2020). Biocontrol of tomato bacterial wilt by the new strain Bacillus velezensis FJAT-46737 and its lipopeptides. BMC Microbiology, 20, 160.

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Culturing Bacillus subtilis in Nutrient Broth

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B. subtilis was purchased from Sigma-Aldrich. B. subtilis was routinely cultured in Nutrient Broth medium (Fisher Scientific, UK).

, & Jebril N.M. (2020). Evaluation of two fixation techniques for direct observation of biofilm formation of Bacillus subtilis in situ, on Congo red agar, using scanning electron microscopy. Veterinary World, 13(6), 1133-1137.

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Isolation and Preservation of Microbial Strains

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Strain FJAT-46737 was isolated from a soil sample from the Huanggang Mountain area, Fujian Province, China, and preserved at the China General Microbiological Culture Collection Center (CGMCC No. 14661). The pathogenic strains R. solanacearum FJAT-91 (CGMCC No. 10692, tomato pathogen), R. solanacearum FJAT-77 (peanut pathogens), Escherichia coli FJAT-301, F. oxysporum f. sp. capsicum FJAT-831, F. oxysporum f. sp. niveum FJAT-30265 and F. oxysporum f. sp. melonis FJAT-9230 were preserved at the Agricultural Bioresources Research Institute, Fujian Academy of Agricultural Sciences, Fujian, China. All the culture media used for bacterial and fungal growth were purchased from AoBoXing Biological Technology Co., Ltd. (Beijing, China). The cultivation substrates used for the pot experiments were purchased from Xiamen Jiang Ping biological Technology Co., Ltd. (Xiamen, China). The reference standards for iturin and surfactin were produced from B. subtilis and purchased from Sigma (St. Louis, MO, USA).

Chen M., Wang J., Liu B., Zhu Y., Xiao R., Yang W., Ge C., & Zheng C. (2020). Biocontrol of tomato bacterial wilt by the new strain Bacillus velezensis FJAT-46737 and its lipopeptides.

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