Bacterial and SCFA Profiling Protocol

Samples were immediately preserved using BioFreeze™ sampling kits (Alimetrics Diagnostics Ltd., Espoo, Finland) following the recommended protocol by the manufacturer. The analysis of dominant bacterial species as well as SCFA profiles was performed by Alimetrics Diagnostics Ltd. Samples were washed and centrifuged for collection of bacterial cells. The cell walls were then disrupted, and chromosomal DNA was extracted and purified for use in quantitative real-time PCR analyses. The SCFA profiles were analyzed by gas chromatography (Agilent Technologies, Santa Clara, CA, USA) using pivalic acid (Sigma-Aldrich, St. Louis, MO, USA) as an internal standard. The chromatography procedure which used a glass column packed with 80/120 Carbopack B-DA/4% Carbowax stationary phase, helium as a carrier gas, and a flame ionization detector has been described previously by Apajalahti et al. [31 (link)].

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, & Adewole D. (2020). Effect of Dietary Supplementation with Coarse or Extruded Oat Hulls on Growth Performance, Blood Biochemical Parameters, Ceca Microbiota and Short Chain Fatty Acids in Broiler Chickens. Animals : an Open Access Journal from MDPI, 10(8), 1429.

Publication 2020

pivalic acid

Bacterial Carbowax Cell walls Cells Chromatography Chromosomal Diagnostics Flame ionization Gas chromatography Helium Pivalic acid Quantitative real time pcr analyses

Corresponding Organization : Dalhousie University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Dominant bacterial species
SCFA profiles

control variables

Sample preservation using BioFreeze™ sampling kits (Alimetrics Diagnostics Ltd., Espoo, Finland)
Analysis of dominant bacterial species and SCFA profiles performed by Alimetrics Diagnostics Ltd.
Washing and centrifugation of samples for collection of bacterial cells
Cell wall disruption and chromosomal DNA extraction and purification for quantitative real-time PCR analyses
SCFA profile analysis by gas chromatography (Agilent Technologies, Santa Clara, CA, USA) using pivalic acid (Sigma-Aldrich, St. Louis, MO, USA) as an internal standard
Chromatography procedure using a glass column packed with 80/120 Carbopack B-DA/4% Carbowax stationary phase, helium as a carrier gas, and a flame ionization detector

controls

Pivalic acid (Sigma-Aldrich, St. Louis, MO, USA) used as an internal standard for SCFA profile analysis

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