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MB Sample ID: SA183497

Local Sample ID:	15_1H_11Abril2018
Subject ID:	SU002023
Subject Type:	Mammal
Subject Species:	Hydrochoerus hydrochaeris
Gender:	Female

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Subject:

Subject ID:	SU002023
Subject Type:	Mammal
Subject Species:	Hydrochoerus hydrochaeris
Gender:	Female

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
15_1H_11Abril2018	SA183497	FL022349	Rectal 3.1	group

Collection:

Collection ID:	CO002016
Collection Summary:	This study was carried out in strict accordance with the Animal Management Rule of the Brazilian Ministry of Environment (Sisbio 59826-1). The samples were obtained from three euthanized female animals in Tatuí/São Paulo State, Brazil (September 2017) as a measure of management of Rocky Mountain Spotted Fever (RMSF) hosts. After euthanasia, 20 g of intestinal contents were collected from the cecum and recto of each animal. All samples were placed in sterile containers and immediately frozen in liquid nitrogen. Samples were kept at −80 °C until processing.
Sample Type:	Cecum

Treatment:

Treatment ID:	TR002035
Treatment Summary:	Approximately 30 mg of dry cecal and rectal contents from three female animals were used for NRM-based metabolomics analysis

Sample Preparation:

Sampleprep ID:	SP002029
Sampleprep Summary:	Approximately 30 mg of dry cecal and rectal contents, and 300 μL of solution 2:1 (methanol: chloroform) were mixed and sonicated for 1 min (4 cycles of 15 sec with intervals of 10 sec) and placed at 4 °C for 15 min. Next, 300 μL of solution 1:1 (methanol: ultrapure water) was added, followed by centrifugation at 16,000 g and 4 °C for 20 min. The supernatant was transferred to a new tube and were dried in CentriVap Solvent System (Labconco Corporation). Samples were diluted to 630 μL by addition of D2O, 70 μL of sodium phosphate buffer (final concentration 0.1 M) containing dimethyl-silapentane-sulfonate (final concentration 0.5 mM) for NMR chemical shift reference and concentration calibration. The samples were filtrated in a syringe filter with a 0.22 µm pore size hydrophilic polyethersulfone (PES) membrane.

Sampleprep ID:

SP002029

Sampleprep Summary:

Approximately 30 mg of dry cecal and rectal contents, and 300 μL of solution 2:1 (methanol: chloroform) were mixed and sonicated for 1 min (4 cycles of 15 sec with intervals of 10 sec) and placed at 4 °C for 15 min. Next, 300 μL of solution 1:1 (methanol: ultrapure water) was added, followed by centrifugation at 16,000 g and 4 °C for 20 min. The supernatant was transferred to a new tube and were dried in CentriVap Solvent System (Labconco Corporation). Samples were diluted to 630 μL by addition of D2O, 70 μL of sodium phosphate buffer (final concentration 0.1 M) containing dimethyl-silapentane-sulfonate (final concentration 0.5 mM) for NMR chemical shift reference and concentration calibration. The samples were filtrated in a syringe filter with a 0.22 µm pore size hydrophilic polyethersulfone (PES) membrane.

Analysis:

MB Sample ID:	SA183497
Analysis ID:	AN003167
Laboratory Name:	Brazilian Center for Research in Energy and Materials (CNPEM)
Analysis Type:	NMR
Num Factors:	6
Num Metabolites:	49
Units:	μM

NMR:

NMR ID:	NM000217
Analysis ID:	AN003167
Instrument Name:	Varian Inova NMR spectrometer (Agilent Technologies Inc.)
Instrument Type:	Other
NMR Experiment Type:	1D-1H
Spectrometer Frequency:	599.84 MHz