Summary of Study ST003440

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002123. The data can be accessed directly via it's Project DOI: 10.21228/M86G0K This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study ID	ST003440
Study Title	Non-targeted metabolomic analysis of Enterobacter hormaechei_B17 (Eh_B17),Enterobacter sichuanensis_B36(Es_B36), and Enterobacter hormaechei_B56 (Eh_B56)
Study Summary	All samples were acquired by the LC-MS system according to the machine instructions. Firstly, all chromatographic separations were performed using an UPLC system (SCIEX). An XBridge BEH C18 column (3.5um,2.1mm × 100mm, Waters) was used for the reversed-phase separation. The column oven was maintained at 50°C. The flow rate was 0.3 mL/min and the mobile phase consisted of solvent A (water containing 0.1% formic acid) and solvent B (acetonitrile containing 0.1% formic acid). The gradient conditions were set as follows: 0-0.5 min, 5% phase B; 0.5-5 min, 5% to 80% phase B; 5-7 min, 80%-100% phase B; 7-8 min,100% phase B; 8-8.1 min, 100%-5% phase B; 8.1-10 min, 5% phase B. The injection volume for each sample was 10 μL.A high-resolution tandem mass spectrometer TripleTOF5600 (SCIEX) was used to detect metabolites eluted from the column. The Q-TOF was operated in both positive ion and negative ion modes. For positive ion mode, the capillary voltages were set to 5 kV. For negative ion mode, the capillary voltages were set to -4.5 kV. Mass spectrometry data were acquired in IDA mode. The TOF mass range was 50 to 1200 Da. The top 8 precursors were fragmented for MS/MS detection. In addition, a quality control sample (pool of all samples) was acquired after every 10 samples to evaluate the stability of the LC-MS throughout the acquisition.
Institute	Institute of Zoology, Chinese Academy of Sciences
Last Name	Zou
First Name	Zhen
Address	1 Beichen West Road, Chaoyang District, Beijing, Beijing, 100101, China
Email	zouzhen@ioz.ac.cn
Phone	+86 15010747660
Submit Date	2024-08-27
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2024-09-23
Release Version	1

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Project:

Project ID:	PR002123
Project DOI:	doi: 10.21228/M86G0K
Project Title:	Gut symbiont-derived sphingosine modulates vector competence in Aedes mosquitoes
Project Summary:	The main vectors of Zika virus (ZIKV) and dengue virus (DENV) are Aedes aegypti and Ae. albopictus, with Ae. aegypti being more competent. However, the underlying mechanisms remain unclear. Here, we find Ae. albopictus shows comparable vector competence to ZIKV/DENV with Ae. aegypti by blood-feeding after antibiotic treatment or intrathoracic injection. This suggests that midgut microbiota can influence vector competence. Enterobacter hormaechei_B17 (Eh_B17) is isolated from field-collected Ae. albopictus and conferred resistance to ZIKV/DENV infection in Ae. aegypti after gut-transplantation. Sphingosine, a metabolite secreted by Eh_B17, effectively suppresses ZIKV infection in both Ae. aegypti and cell cultures by blocking viral entry during the fusion step, with an IC50 of approximately 10 μM. A field survey reveals that Eh_B17 preferentially colonizes Ae. albopictus compared to Ae. aegypti. And field Ae. aegypti positive for Eh_B17 are more resistant to ZIKV infection. These findings underscore the potential of gut symbiotic bacteria, such as Eh_B17, to modulate the arbovirus vector competence of Aedes mosquitoes. As a natural antiviral agent, Eh_B17 holds promise as a potential candidate for blocking ZIKV/DENV transmission.
Institute:	Institute of Zoology, Chinese Academy of Sciences
Last Name:	Zou
First Name:	Zhen
Address:	1 Beichen West Road, Chaoyang District, Beijing, Beijing, 100101, China
Email:	zouzhen@ioz.ac.cn
Phone:	+86 15010747660

Subject:

Subject ID:	SU003567
Subject Type:	Bacteria
Subject Species:	Enterobacter sp.

Factors:

Subject type: Bacteria; Subject species: Enterobacter sp. (Factor headings shown in green)

mb_sample_id	local_sample_id	Bacterium	Sample source
SA378912	B17-F6_N-1	Enterobacter hormaechei	bacterial supernatant
SA378913	B17-F6_N-2	Enterobacter hormaechei	bacterial supernatant
SA378914	B17-F6_N-3	Enterobacter hormaechei	bacterial supernatant
SA378915	B17-F6_P-1	Enterobacter hormaechei	bacterial supernatant
SA378916	B17-F6_P-2	Enterobacter hormaechei	bacterial supernatant
SA378917	B17-F6_P-3	Enterobacter hormaechei	bacterial supernatant
SA378918	B56_N	Enterobacter hormaechei	bacterial supernatant
SA378919	B56_P	Enterobacter hormaechei	bacterial supernatant
SA378920	B36_N	Enterobacter sichuanensis	bacterial supernatant
SA378921	B36_P	Enterobacter sichuanensis	bacterial supernatant
SA378922	QC1_N	mixture of B17-F6, B36, and B56	bacterial supernatant
SA378923	QC1_P	mixture of B17-F6, B36, and B56	bacterial supernatant
SA378924	QC2_N	mixture of B17-F6, B36, and B56	bacterial supernatant
SA378925	QC2_P	mixture of B17-F6, B36, and B56	bacterial supernatant

Showing results 1 to 14 of 14

Showing results 1 to 14 of 14

Collection:

Collection ID:	CO003560
Collection Summary:	Enterobacter hormaechei_B17 (B17) and Enterobacter sichuanensis_B36 (B36) were isolated from the midgut of wild Aedes albopictus, while Enterobacter hormaechei_B56 (B56) was isolated from the midgut of lab-adapted Aedes aegypti. The flow-through of Eh_B17 was then separated into six fractions by semi-preparative high performance liquid chromatography (SPHPLC) according to the peak time. The parts were named F1 (0–6.5 min), F2 (6.5–10 min), F3 (10–15.5 min), F4 (15.5–20 min), F5 (20–25.5 min), and F6 (25.5–55 min).
Sample Type:	bacteria

Treatment:

Treatment ID:	TR003576
Treatment Summary:	No treatment

Sample Preparation:

Sampleprep ID:	SP003574
Sampleprep Summary:	B36 and B56 were grown overnight in LB medium at 37°C with an OD600 of 0.6. Then, a 200-μL bacterial solution was applied to LB plates and maintained for 3 d. The bacterial colonies were collected in 1 mL methanol and then put in a water bath at 80°C until the methanol evaporated completely. Samples were reconstituted in 0.5 mL of methanol and centrifuged at 12,000 × g for 3 min to collect the supernatant. The liquid was evaporated using a rotary evaporator, and the dry powder was dissolved in ddH2O to at a concentration of 50 mg/mL. Liquid was filtrated with a membrane filter (0.45 µm) and stored at −20°C for further testing. At least three independent experiments were performed. B17 was grown in LB medium at 37°C until OD600 reached 0.6, and then transferred to 16°C and incubated for 48 h. The bacterial supernatant was collected and concentrated using vacuum freeze-drying (ALPHA LDplus, Germany). The dry powder was resuspended in ddH2O in a 1/20 volume of the original supernatant. The resuspended supernatant was separated into retention (mainly protein) and flow-through (mainly metabolites) by a 3 kDa centrifugal filter. The fluids were stored at −80°C for further experiments.

Sampleprep ID:

SP003574

Sampleprep Summary:

B36 and B56 were grown overnight in LB medium at 37°C with an OD600 of 0.6. Then, a 200-μL bacterial solution was applied to LB plates and maintained for 3 d. The bacterial colonies were collected in 1 mL methanol and then put in a water bath at 80°C until the methanol evaporated completely. Samples were reconstituted in 0.5 mL of methanol and centrifuged at 12,000 × g for 3 min to collect the supernatant. The liquid was evaporated using a rotary evaporator, and the dry powder was dissolved in ddH2O to at a concentration of 50 mg/mL. Liquid was filtrated with a membrane filter (0.45 µm) and stored at −20°C for further testing. At least three independent experiments were performed. B17 was grown in LB medium at 37°C until OD600 reached 0.6, and then transferred to 16°C and incubated for 48 h. The bacterial supernatant was collected and concentrated using vacuum freeze-drying (ALPHA LDplus, Germany). The dry powder was resuspended in ddH2O in a 1/20 volume of the original supernatant. The resuspended supernatant was separated into retention (mainly protein) and flow-through (mainly metabolites) by a 3 kDa centrifugal filter. The fluids were stored at −80°C for further experiments.

Combined analysis:

Analysis ID	AN005655	AN005656
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	ExionUPLC（SCIEX）	ExionUPLC（SCIEX）
Column	Waters XBridge BEH C18 (100 x 2.1mm,3.5um)	Waters XBridge BEH C18 (100 x 2.1mm,3.5um)
MS Type	ESI	ESI
MS instrument type	QTOF	QTOF
MS instrument name	ABI Sciex 5600 TripleTOF	ABI Sciex 5600 TripleTOF
Ion Mode	POSITIVE	NEGATIVE
Units	Peak area	Peak area

Chromatography:

Chromatography ID:	CH004293
Instrument Name:	ExionUPLC（SCIEX）
Column Name:	Waters XBridge BEH C18 (100 x 2.1mm,3.5um)
Column Temperature:	50°C
Flow Gradient:	0–0.5 min, 5% phase B; 0.5–5 min, 5%–80% phase B; 5–7 min, 80%–100% phase B; 7–8 min,100% phase B; 8–8.1 min, 100%–5% phase B; 8.1–10 min, 5% phase B
Flow Rate:	0.3 mL/min
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100% acetonitrile; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS005379
Analysis ID:	AN005655
Instrument Name:	ABI Sciex 5600 TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	A high-resolution tandem mass spectrometer TripleTOF5600（SCIEX）was used to detect metabolites eluted form the column. The Q-TOF was operated in both positive and negative ion modes. For positive ion mode, the capillary voltages were set at 5 kV. For negative ion mode, the capillary voltages were set at -4.5 kV. The mass spectrometry data were acquired in IDA mode. The TOF mass range was from 50 to 1200 Da. For the MS/MS detection, top 8 precursors were fragmented. Furthermore, in order to evaluate the stability of the LC-MS during the whole acquisition, a quality control sample (Pool of all samples) was acquired after every 10 samples.
Ion Mode:	POSITIVE

MS ID:	MS005380
Analysis ID:	AN005656
Instrument Name:	ABI Sciex 5600 TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	A high-resolution tandem mass spectrometer TripleTOF5600（SCIEX）was used to detect metabolites eluted form the column. The Q-TOF was operated in both positive and negative ion modes. For positive ion mode, the capillary voltages were set at 5 kV. For negative ion mode, the capillary voltages were set at -4.5 kV. The mass spectrometry data were acquired in IDA mode. The TOF mass range was from 50 to 1200 Da. For the MS/MS detection, top 8 precursors were fragmented. Furthermore, in order to evaluate the stability of the LC-MS during the whole acquisition, a quality control sample (Pool of all samples) was acquired after every 10 samples.
Ion Mode:	NEGATIVE