Samples of feed, ruminal contents, and feces were processed and analyzed using the same techniques for both experiments. Samples were dried at 55 °C in a forced-air oven for 96 h and allowed to air equilibrate for 24 h for the determination of partial dry matter (DM). Samples were then ground to pass through a 1-mm screen (No. 4 Wiley Mill, Thomas Scientific, Swedesboro NJ). Ground samples were dried at 105 °C for final DM determination. Organic matter (OM) was determined as the loss in DM weight following combustion for 8 h at 450 °C in a muffle furnace. Acid detergent fiber (ADF) analysis was performed using an Ankom Fiber Analyzer (Ankom Technology Corp., Macedon, NY) with sodium sulfite and amylase omitted and without correction for residual ash. Sample ADIA was measured as the remaining DM upon the combustion of ADF residue in a muffle furnace. The energy content of samples was determined by direct calorimetry using a Parr 6300 Calorimeter (Parr Instrument Company, Moline, IL). Samples of ruminal fluid were thawed and centrifuged at 20,000 × g for 20 min. Volatile fatty acid concentrations were measured using a gas chromatograph with methods described by Vanzant and Cochran (1994) (link).
6300 calorimeter
Manufactured by Parr
Sourced in United States, Israel
The 6300 Calorimeter is a laboratory equipment used to measure the heat of reaction or the heat capacity of a substance. It provides precise and accurate calorimetric measurements to determine the energy changes in chemical or physical processes.
Automatically generated - may contain errors
Lab products found in correlation
Fiber analyzer, by ANKOM Technology (6 mentions)
Uv 1201, by Shimadzu (2 mentions)
Ankom fiber analyzer, by ANKOM Technology (1 mentions)
Autosampler, by Hewlett-Packard (1 mentions)
Ankom xt 10 extractor, by ANKOM Technology (1 mentions)
Z 5000, by Hitachi (1 mentions)
L 8900, by Hitachi (1 mentions)
Automatic amino acid analyzer, by Hitachi (1 mentions)
Atomic absorption spectrometer, by Hitachi (1 mentions)
A220 fiber analyzer, by ANKOM Technology (1 mentions)
17 protocols using 6300 calorimeter
1
Feed, Ruminal, and Fecal Sample Analysis
2
SCFA Analysis and Digestibility Assay
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At T120, the end of the experiment, 2 mL of the content of each bioreactor was recovered. Then, 4 mL of H2SO4 1.5% (Fisher Scientific, Waltham, MA, USA) were added, then the samples were vortexed. The samples were kept at −20 °C until SCFA analysis was performed. The frozen samples were homogenized and then centrifuged at 10,000 rpm for 15 min at 4 °C. The supernatant was analyzed on a Hewlett Packard 6890N gas chromatograph (Agilent Technologies, Wilmington, DE, USA) equipped with a flame ionization detector and an autosampler (Hewlett, Packard, Avondale, PA, USA) for volatile fatty acid.
The rest of the content of each bioreactor (approximately 40 mL) was individually flash-frozen and kept at −80 °C for digestibility analysis. These samples were analyzed for gross energy (GE, Parr 6300 Calorimeter, Parr Instrument Company, Moline, IL, USA), crude protein (Kjeldahl, method 976.05; nitrogen × 6.25) and dry matter (method 930.15).
The rest of the content of each bioreactor (approximately 40 mL) was individually flash-frozen and kept at −80 °C for digestibility analysis. These samples were analyzed for gross energy (GE, Parr 6300 Calorimeter, Parr Instrument Company, Moline, IL, USA), crude protein (Kjeldahl, method 976.05; nitrogen × 6.25) and dry matter (method 930.15).
3
Dietary Lipid Composition and CFAM Supplementation
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Table 1 presents the formulation of the four (4) diet groups based on a combination of the type of oil (canola or soybean) and the dose of CFAM (0.0% and 0.5% of total dietary lipids): canola oil (CO), canola and CFAM (CC), soybean oil (SO), and soybean and CFAM (SC). Fatty acid profiles of the diets were determined by gas chromatography (GC) and are also presented in Table 1 . The diets were formulated to be isoenergetic, isolipidic and isonitrogenous. Total energy in diets was determined with an adiabatic Parr 6300 calorimeter (Parr Instrument Company, Moline, IL, USA) and was similar among the four diets (CO, 19.5 kJ/g; CC, 19.4 kJ/g; SO, 19.5 kJ/g; SC, 19.5 kJ/g). Dietary protein content was determined by combustion (Dumas method) using a LECO FP‐528 apparatus (LECO Corporation, St. Joseph, MI, USA) and was also found similar between the diets (CO, 19.3% [w/w]; CC, 19.3% [w/w]; SO, 18.9% [w/w]; SC, 19.7% [w/w]). Dietary fat content was measured with an ANKOMXT10 Extractor (ANKOM Technology, Macedon, NY, USA) and was similar between the diets (CO, 10.1% [w/w]; CC, 10.3% [w/w]; SO, 10.0% [w/w]; SC, 10.2% [w/w]).
4
Comprehensive Feed Composition Analysis
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Samples were analyzed for GE via an adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter; Parr Instrument Company, Moline, IL, USA). Other analysis including DM, CP, and ash analyzed according to AOAC [12 ]. The ether extract (EE) was analyzed by a previous method [13 (link)]. Total dietary fiber (TDF) and insoluble dietary fiber (IDF) in SBP or corn grain were also determined according to AOAC [12 ]. The concentration of soluble dietary fiber (SDF) was calculated as the difference between TDF and IDF. Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined using filter bags and Fiber Analyzer equipment (Fiber Analyzer; ANKOM Technology, Macedon, NY, USA) following a modification of the procedures [14 (link)]. The concentration of NDF was analyzed using heat-stable α-amylase and sodium sulfite without correction for insoluble ash. Organic matter (OM) was calculated as the difference between DM and ash. All chemical analyses were conducted in duplicates.
5
Detailed Chemical Analysis of Feedstuffs
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Samples of ingredients and diets were ground through a 1-mm screen, and then analyzed for DM (AOAC, 2007 ; method 930.15), CP (AOAC, 2007 ; method 976.05), and ash (AOAC, 2007 ; method 942.15). The neutral detergent fiber and acid detergent fiber were determined using a fiber analyzer (Ankom Technology, Macedon, NY) according to Van Soest et al. (1991) (link) with heat-stable α-amylase and sodium sulfite and expressed inclusive of residual ash. The gross energy was determined using an automatic adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter; Parr Instrument Company, Moline, IL). The TDF and IDF were analyzed using AOAC (2007) methods 985.29 and 991.43, respectively. The SDF was calculated as the difference between TDF and IDF.
6
Nutrient Composition and Digestibility Analysis
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Ingredients, diets and excreta samples were ground through a 1-mm screen and then analyzed for dry matter (DM; AOAC, 2007; method 930.15), crude protein (CP; AOAC, 2007; method 976.05) and ash (AOAC, 2007; method 942.15) [17 ]. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined using fiber analyzer (Ankom Technology, Macedon, NY, USA) according to Van Soest, et al. [18 (link)]. The gross energy (GE) was determined using an automatic adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter, Moline, IL, USA). The total dietary fiber (TDF) and IDF were analyzed using AOAC (2007) [17 ] methods 985.29 and 991.43, respectively. The SDF was calculated as the difference between TDF and IDF. The chromium (Cr) content was measured using anatomic absorption spectrophotometer (Z-5000; Hitachi, Tokyo, Japan) according to the procedure of Williams et al. [19 (link)]. The apparent total tract digestibility (ATTD) values were calculated by the equation as follows: ATTD (%) = [1 − (Crdiet × Nutrientexcreta)/(Crexcreta × Nutrientdiet)] × 100%, where Crdiet represents the chromium content in diet, and Crexcreta represents the chromium content in excreta. Nutrientexcreta represents the nutrient content in excreta, and Nutrientdiet represents the nutrient content in diet.
7
Protocol for Nutrient Analysis of Feed and Excreta
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Feed and dried excreta samples were ground to pass through a 40-mesh screen and mixed thoroughly. Samples were analyzed in duplicates for dry matter (DM) and crude protein (CP) according to the standard methods of AOAC (1990) . Gross energy (GE) content was measured using an automatic adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter; Moline, IL, USA). Chromium concentration in feed and excreta was analyzed using an atomic absorption spectrophotometer (Hitachi Z-2000 Automatic Absorption; Spectrophotometer, Tokyo, Japan) according to the method of Williams et al. (1962) .
8
Nutrient Composition Analysis of Diets and Rice Bran
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Dry matter (DM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), ash, calcium and phosphorus content in the diets and rice bran samples were analyzed according to the procedures of the Association of Official Analytical Chemists (AOAC, 2000 ). Acid hydrolyzed ether extract (AEE) was determined by acid hydrolysis using 3 N HCL (Sanderson, 1986 ) followed by crude fat extraction with petroleum ether (AOAC, 2005 ) using an Extraction System (Extractor, Ankom Technology, Macedon, NY, USA). The gross energy (GE) of feces, urine, diets and rice bran samples were measured by an automatic adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter, Moline, IL, USA) with benzoic acid as a standard. Before the GE of urine was measured, a 4 mL sample of urine was injected onto 2 filter papers in a special crucible, and dried for 8 h in a 65°C drying oven. The starch content was determined after converting starch to glucose using an enzyme assay kit (Megazym International Ireland, Wicklow, Ireland).
9
Analytical Methods for Feed Ingredients
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Ingredients and diets were ground through a 1-mm screen, and then analyzed for dry matter (DM; AOAC, 2007 ; method 930.15), crude protein (CP; AOAC, 2007 ; method 976.05) and ash (AOAC, 2007 ; method 942.15). Neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined using a fiber analyzer (Ankom Technology, Macedon, NY, USA) according to the method described by Van Soest et al. (1991) (link). The gross energy (GE) was determined using an automatic adiabatic oxygen bomb calorimeter (Parr 6300 Calorimeter, Moline, IL, USA). TDF and IDF were analyzed using AOAC (2007) methods 985.29 and 991.43, respectively. SDF was calculated as the difference between TDF and IDF. Amino acids, except methionine, tryptophan and cystine, were assayed using ion-exchange chromatography with an Automatic Amino Acid Analyzer (L-8900, Automatic Amino Acid Analyzer; Hitachi, Tokyo, Japan) after hydrolyzing with 6 mol/L HCl at 110 °C for 24 h. Cystine was determined as cysteic acid and Met as methionine sulphone after peroxidation with performic acid and pre-column derivation using phenylisothiocyanate (L-8900, Automatic Amino Acid Analyzer; Hitachi, Tokyo, Japan). Tryptophan was determined after hydrolyzing with 4 mol/L LiOH at 110 °C for 22 h using high performance liquid chromatography (Agilent1200 Series; Aligent, Santa Clara, CA, USA).
10
Comprehensive Nutritional Analysis of Rapeseed Meal
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Ingredients, diets and feces were analyzed for dry matter (DM; method 930.15, [10 ]) and organic matter (OM) was calculated as DM minus ash content. Crude protein (CP, method 984.13 [10 ]), ash (method 942.05 [10 ]), and EE [11 (link)] were analyzed in ingredients, diets, and feces. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) were determined using filter bags and Fiber Analyzer equipment (Fiber Analyzer, Ankom Technology, Macedon, NY, USA) following a modification of the procedure of Van Soest et al [12 (link)]. Total glucosinolate concentration was analyzed in ingredients according to Daun and McGregor [13 ]. The gross energy (GE) in the five RSMs, diets, feces, and urine samples were analyzed using an isoperibol calorimeter (Parr 6300 Calorimeter, Moline, IL, USA) with benzoic acid as a standard. The five RSMs and diets were also analyzed for total starch by the glucoamylase procedure (Method 948.02 [10 ]).
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