Near-infrared calibration models for estimating volatile fatty acids and methane production from in vitro rumen fermentation of different total mixed rations

Abstract

Volatile fatty acids (VFA) and methane (CH₄) are the major products of rumen fermentation. The VFA are considered an energy source for the animal and rumen microbiota, and CH₄ (which is released by eructation) is considered an energy loss. Quantification of these fermentation products is fundamental for the evaluation of feeds and diets, and provides important information regarding the use of nutrients by ruminants. Near-infrared (NIR) spectroscopy is increasingly used for the evaluation of animal feeds because it is rapid, nondestructive, noninvasive, and inexpensive; does not require reagents; and the results are reproducible. The aim of this study was to develop NIR calibration models for estimating the production of VFA (acetic, propionic, butyric, valeric, isovaleric, and isobutyric acids), total gas, and CH₄ using in vitro gas production tests with buffered rumen inoculum throughout fermentation. Fifty-four total mixed rations (TMRs) were examined, and rumen fluid was manually collected from 2 dry Holstein dairy cows that had ruminal fistulas and were fed at maintenance energy levels. Then, 30 mL of buffered rumen fluid was incubated in bottles with ~220 mg of TMR. The total gas, VFA, and CH₄ were measured after 2, 5, 9, 24, 30, 48, and 72 h of rumen incubation for each TMR. The VFA were measured on 32 randomly selected TMR. In particular, 7 bottles were used for each TMR, one for each incubation time. Methane was measured in the headspace and VFA were measured in the buffered rumen fluid. The bottles were considered experimental units for calibration purposes. The production of CH₄ was quantified from the bottle headspaces by gas chromatography, and total gas production was measured using a pressure transducer at each incubation time. Two aliquots of the fermented liquids were sampled by opening the bottles at each incubation time, and (1) the concentrations of VFA were determined by gas chromatography or (2) spectra were obtained from Fourier-transform NIR spectroscopy. The data were randomly divided into calibration and validation data sets. The average concentrations of acetic acid (45.30 ± 11.92 and 43.86 ± 11.93 mmol/L), propionic acid (14.97 ± 6.08 and 14.38 ± 6.56 mmol/L), butyric acid (8.47 ± 3.47 and 8.65 ± 3.79 mmol/L), total gas (111.34 ± 81.90 and 116.46 ± 82.44 mL/g of organic matter), and CH₄ (9.65 ± 9.45 and 10.35 ± 9.33 mmol/L) were similar in the 2 data sets. The best calibration models were retained based on the coefficient of determination (R²) and the ratio of prediction to deviation (RPD). The R² values for prediction of VFA ranged from 0.69 (RPD = 3.28) for valeric acid to 0.94 (RPD = 4.20) for acetic acid. The models also provided good predictions of CH₄ (R² = 0.89, RPD = 3.05) and cumulative gas production (R² = 0.91, RPD = 3.30). The models described here precisely and accurately estimated the production of CH₄ and VFA during in vitro rumen fermentation tests. Validations at additional laboratories may provide more robust calibrations.

Summary: Diverse disciplines of science use near-infrared (NIR) spectroscopy for chemical analysis because it is rapid and inexpensive, and it provides reproducible results. Many animal scientists perform in vitro rumen fermentation tests to measure the production of volatile fatty acids (VFA) and methane (indicative of energy loss) from different feeds or diets. We used NIR spectroscopy to develop calibration models that predict the production of different VFA (acetic, propionic, butyric, valeric, isovaleric, and isobutyric acids), total gas, and methane from in vitro rumen fermentation of different silage-based diets. Our models provided reliable estimates of these rumen fermentation products.

Figure 1

Selection of latent variables for each partial least squares model: (a) acetic acid; (b) propionic acid; (c) butyric acid; (d) isobutyric acid; (e) valeric acid; (f) isovaleric acid; (g) methane; and (h) total gas.