Advanced Vibrational Molecular Spectroscopy Research Articles

Using vibrational molecular spectroscopy to reveal carbohydrate molecular structure properties of faba bean partitions and faba bean silage before and after rumen incubation in relation to nutrient availability and supply to dairy cattle.

To our knowledge, there is limited study on the relationship between the molecular structure of feed and nutrient availability in the ruminant system. The objective of this study is to use advanced vibrational molecular spectroscopy (attenuated total reflection [ATR]-Fourier transform infrared [FT/IR]) to reveal carbohydrate molecular structure properties of faba bean partitions (stem, leaf, whole pods [WP], and whole plant) and faba bean silage before and after rumen incubation in relation to nutrient availability and supply to dairy cattle. The study included the correlation between carbohydrate-related spectral profiles and chemical profiles, feed energy values, Cornell Net Carbohydrate and Protein System carbohydrate fractions, and rumen degradation parameters of faba bean samples (whole crop, stem, leaf, WP, and silage) before and after rumen incubation. FTIRspectra of faba bean sample before and after 12 and 24 h rumen incubations were collected with JASCO FT/IR-4200 with ATR at mid-IR range (ca. 4000-700 cm-1 ) with 128 scans and at 4 cm-1 resolution. The univariate molecular spectral analysis was carried out using OMNIC software. The results show that ATR-FT/IR spectroscopic technique could detect the change of microbial digestion to carbohydrate-related molecular structure. The spectral parameters of feed rumen incubation residues had a stronger correlation with less degradable carbohydrate fractions (neutral detergent fiber, acid detergent fiber,acid detergent lignin, hemicellulose, and cellulose) while spectral profiles of original faba samples had a stronger correlation with easily degradable carbohydrate fractions (starch). In conclusion, rumen degradation of carbohydrate contents can be reflected in the change of its molecular spectral profiles. The study shows that vibrational molecular spectroscopy (ATR-FT/IR) shows high potential as a fast analytical tool to evaluate and predict nutrient supply in the ruminant system.

Using advanced vibrational molecular spectroscopy to detect moist heating induced protein structure changes in cool-climate adapted barley grain.

Different techniques have been applied in feed processing to improve ruminal degradation and nutrient utilization in ruminant. There are limited studies investigating how moist heating process impacts barley protein utilization and internal molecular structures. The objectives of this study were to investigate: 1) how moist heating affects barley protein chemical profiles and Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation parameters, and predicted intestinal protein supply and feed milk value; 2) how moist heating affects protein molecular spectral features; and 3) the relationship between protein molecular structure spectral features and protein chemical profiles and metabolic characteristics. The barley variety CDC cowboy samples collected from the research farm during two consecutive years were used. Half of each sample was kept as raw and the other half underwent moist heating. The advanced molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley protein molecular structure spectral features. It was found that moist heating decreased the fractions of soluble protein and increased the moderately degradable protein and ingestible protein fractions. This further resulted in the changes of in situ rumen degradation parameters and intestinal protein digestion characteristics. The protein molecular structure spectral features detected by using ATR-FTIR spectroscopy can be used as potential predictors for protein related chemical and metabolic parameters.

Using vibrational molecular spectroscopy to detect moist heating induced carbohydrates structure changes in cool-climate adapted barley grain

There is limited research to study how moist heating affects internal structure of barley grain on a molecular basis. The objectives of this study were to use vibrational molecular spectroscopy: 1) to determine the moist heating induced changes of barley carbohydrate (CHO) structure on a molecular basis, 2) to study the effects of moist heating on CHO chemical profiles, Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation, and predicted intestinal carbohydrate supply of barley grain; and 3) to reveal the association between molecular structure spectral features and CHO related metabolic characteristics. Barley samples (CDC cowboy) were collected from Kernen Crop Research Farm (Saskatoon, Canada) during two consecutive years. Half of each sample was kept as raw barley and the other half underwent moist heating (autoclaving at 120 °C for 60 min). The molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley CHO related molecular structure spectral features. Moist heating did not affect carbohydrate related chemical profiles and CNCPS subfractions but it decreased rumen degradable carbohydrate. Rumen undegradable and intestinal digestion of CHO subfractions were not affected by moist heating. The advanced vibrational molecular spectroscopy can be used to detect carbohydrate molecular spectral features. Nutrient utilization prediction using molecular spectral characteristics is warranted and further investigation is encouraged.

Using advanced vibrational molecular spectroscopy (ATR-Ft/IRS) to study heating process induced changes on protein molecular structure of biodegradation residues in cool-climate adapted faba bean seeds: Relationship with rumen and intestinal protein digestion in ruminant systems.

The objective of this study was to evaluate the effects of heating process on protein molecular structure from ruminal degradation residues in cool-climate adapted faba bean seeds in relation to crude protein (CP), in situ degradation kinetics, rumen protein degradation and intestinal protein digestion parameters in dairy cows. Seeds of six faba bean varieties with low (Snowbird, Snowdrop, 219_16) and normal tannin (Fatima, 346_10, SSNS_1) were collected from three different locations, and were heated 3min by microwave irradiation (MI, dry heating) or heated 1h by steam pressure toasting (SP, moist heating) or kept raw as a control. Heat treated samples were used for rumen incubating 24, 12, 8, 4, 2, 0h(s) in two replicate runs and then residues from 12h of rumen degradation were used for three steps in vitro technique for determining intestinal protein digestion. Attenuated total reflectance Fourier transforms infrared spectroscopy (ATR-Ft/IRS) was used for analyzing protien molecular structure of residual faba bean seeds. The results showed that SP increased the intensities of amide I, amide II, α-helix and β-sheet but decreased amide I to amide II height and area ratio, α-helix to β-sheet height ratio from 12 and 24h of ruminal degradation, and MI decreased all the intensities of amide I, amide II, α-helix and β-sheet and ratios except amide I to amide II area ratio of residues from 24h of ruminal degradation. Additionally, the intensities of amide I, amide II, α-helix and β-sheet had a unique pattern of increasing first and then decreasing with the increasing ruminal digestion time for SP treatment, while amide I to amide II height and area ratio, α-helix to β-sheet height ratio were declining. For the MI groups, this pattern was not observed and the intensities were rather consistent across the digestion process. Rumen protein degradation parameters including rumen bypass crude protein (BCP) or rumen undegradable protein (RUP) and rumen degradable protein (RDP) closely correlated with protein molecular structure of to peak heights, areas and ratios. Regression equations based on residual protein molecular structure presented a good estimation power for soluble fraction (S, R2=0.79), degradable fraction (D, R2=0.805), BCP (R2=0.941), RUP (R2=0.941) and RDP (R2=0.811). Overall, heat-induced changes in rumen residual protein molecular structures were related to CP, in situ degradation kinetics, rumen protein degradation and rumen protein digestion parameters.

Using advanced vibrational molecular spectroscopy (ATR-Ft/IRS and synchrotron SR-IMS) to study an interaction between protein molecular structure from biodegradation residues and nutritional properties of cool-climate adapted faba bean seeds.

The objective of this study was to use advanced vibrational molecular spectroscopy (ATR-Ft/IRS) to study an interaction between legume protein molecular structure from biodegradation residues and nutritional properties of newly developed genotypes of cool-climate adapted faba bean seed with low and normal tannin levels grown in western Canada. Protein molecular structures including amide I, II areas and peak heights, α-helix and β-sheet peak heights in rumen biodegradation residues were determined by using attenuated total reflectance Fourier transform infrared molecular spectroscopy (ATR-FTIR). The nutritional properties were determined which included chemical and nutrient profiles, in situ rumen degradation kinetics, rumen protein degradation, and intestinal protein digestion in the newly developed genotypes of faba bean seeds with low and normal tannin levels. The results showed that the spectral intensity of faba bean varieties with a normal tannin level in rumen biodegradation residues was greater (P<0.05) than that with a low tannin level. The spectral intensity of amide I, II areas and peak heights, α-helix and β-sheet peak heights in all genotypes (except the variety of Snowdrop) in biodegradation residuals of faba bean seeds had a unique pattern with increasing first and then decreasing with the increasing of rumen incubation time. The molecular structures of protein (α-helix, ratio of α-helix to β-sheet height and amide I to II area, R2>0.6) were associated with in situ degradation kinetics - soluble (S) and potential degradable fractions (D) and rumen protein degradation- bypass or undegraded protein (BCP or RUP). The molecular spectral parameters in the FTIR fingerprint region didn't form cluster among different genotypes in residual faba bean seeds in 12h and 24h incubation, which indicate they had similar protein molecular structures after incubation. In conclusion, there was an interaction between protein molecular structure from biodegradation residues and nutritional properties of newly developed cool-climate adapted faba bean seeds with normal and low level of tannin. The cool-climate adapted genotype had an impact on the protein molecular structure, and the protein utilization and metabolism were predictable from protein spectral molecular structures after rumen biodegradation with ATR-Ft/IRS spectroscopy.

Interactive association between processing induced molecular structure changes and nutrient delivery on a molecular basis, revealed by cutting-edge vibrational biomolecular spectroscopy

BackgroundThis study was conducted to determine protein molecular structure profiles and quantify the relationship between protein structural features and protein metabolism and bioavailability of blend pelleted products (BPP) based on co-products (canola or carinata) from processing with different proportions of pulse pea screenings and lignosulfonate chemical compound.MethodThe protein molecular structures were determined using the non-invasive advanced vibrational molecular spectroscopy (ATR-FT/IR) in terms of chemical structure and biofunctional groups of amides (I and II), α-helix and β-sheet.ResultsThe results showed that increasing the level of the co-products in BPP significantly increased the spectral intensity of the amide area and amide height. The products exhibited similar protein secondary α-helix to β-sheet ratio. The protein molecular structure profiles (amides I and II, α-helix to β-sheet) were highly associated with protein degradation kinetics and intestinal digestion. In conclusion, the non-invasive vibrational molecular spectroscopy (ATR-FT/IR) could be used to detect inherent structural make-up characteristics in BPP.ConclusionThe molecular structural features related to protein biopolymer were highly associated with protein utilization and metabolism.

Protein molecular structure in relation to predicted biodegradation and nutrient supply of feedstocks and co-products from bio-oil processing with CNCPS system: Comparison Crusher Plants within Canada and within China as well as between Canada and China

Recently, rapid non-invasive advanced vibrational molecular spectroscopy has been developed to detect tissue inherent molecular structure. However, this advanced technique is seldom known by animal scientists and seldom used to study feed molecular structure in relation to nutrient utilization and availability in animals. The objectives of this study were to 1) detect the interactive association between the processing-induced protein molecular structure changes and biodegradation and intestinal digestion in ruminants using advanced vibrational molecular spectroscopy, and 2) develop a model to predict protein and carbohydrate degradation and digestions based on feed inherent protein molecular structure. The feed used for this study were various sources of feedstock (oil seeds) and co-products (canola meal, canola pellets) from ten different crushing plants in Canada and China with multi-source samples from each crushing plant. The predictable protein and carbohydrate degradable sub-fractions and nutrient supply of sub-fractions to dairy cows were evaluated with the updated Cornell Net Carbohydrate and Protein System (CNCPS 6.5). The protein molecular structure was revealed using advanced vibrational molecular spectroscopy (FT/IR-ATR). The molecular structure spectral analyses were carried out using uni- and multi-variate molecular spectral analyses. The model variable selection was carried out using SAS with a Stepwise option. The results showed that protein molecular structure spectral characteristics could be used to predict CNCPS protein and carbohydrate nutrient supply to dairy cows. Strong positive correlation was found in canola meal between rumen degraded PA2 and amide I area (r = 0.67, P < 0.001), amide I height (r = 0.73, P < 0.001), and α-helix height (r = 0.83, P < 0.001). RDPB2 was negatively correlated with α-helix height (r = −0.69, P < 0.001). Moreover, TRDC was negatively correlated with amide I area (r=-0.64, P < 0.001), amide I height (r = −0.65, P < 0.001), and α-helix height (r = −0.62, P < 0.001). It was concluded that protein and carbohydrate degradable and undegradable sub-fractions had interactive associations with the processing-induced protein molecular structure changes in co-products from bio-oil processing.

Determine effect of pressure heating on carbohydrate related molecular structures in association with carbohydrate metabolic profiles of cool-climate chickpeas using Globar spectroscopy

Grain has been heat-processed to alter rumen degradation characteristics and improve nutrient availabilities for ruminants. However, limited study was found on internal structure changes induced by processing on a molecular basis. The objectives of this study were to use advanced vibrational molecular spectroscopy to: (1) determine the processing induced carbohydrate (CHO) structure changes on a molecular basis, (2) investigate the effect of pressure heating on changes of CHO chemical profiles, CHO subfractions in cool-climate CDC Chickpea varieties, and (3) to reveal the association between carbohydrates related molecular spectra with carbohydrate metabolic profiles. The cool-climate CDC chickpea varieties with multisource were pressure heated in an autoclave at 120 °C for 60 min; and FTIR vibrational spectroscopy was used to detect the molecular spectra. Molecular spectroscopic results showed that compared to raw chickpea varieties, autoclave heating induced changes in both total CHO (region and baseline ca. 1186–946 cm−1) and structural CHO (STCHO, region and baseline ca. 1482–1186 cm−1), except for cellulosic compounds (CELC, region and baseline ca. 1374–1212 cm−1). The CHO chemical profile and rumen degradation results showed that autoclave heating decreased rumen degradable, undegradable and intestinal digestible sugar (CA4) content, but increased available fiber (CB3) content, without affecting available energy of chickpeas. The changes of CHO molecular spectra in chickpea varieties were strongly correlated with CHO chemical profiles, CHO subfractions, and CHO rumen degradation characteristics. Moreover, the regression analysis showed that STCHO peak 1 height could be used to predict sugar content, its rumen degradability and digestibility of chickpeas. Our results suggest that autoclave heating markedly changes sugar and fiber degradation characteristics. The carbohydrate molecular spectral profiles are associated with carbohydrate metabolic profiles in raw and pressure heated cool-climate chickpeas.

Using vibrational molecular spectroscopy to reveal association of steam-flaking induced carbohydrates molecular structural changes with grain fractionation, biodigestion and biodegradation

Advanced vibrational molecular spectroscopy has been developed as a rapid and non-destructive tool to reveal intrinsic molecular structure conformation of biological tissues. However, this technique has not been used to systematically study flaking induced structure changes at a molecular level. The objective of this study was to use vibrational molecular spectroscopy to reveal association between steam flaking induced CHO molecular structural changes in relation to grain CHO fractionation, predicted CHO biodegradation and biodigestion in ruminant system. The Attenuate Total Reflectance Fourier-transform Vibrational Molecular Spectroscopy (ATR-Ft/VMS) at SRP Key Lab of Molecular Structure and Molecular Nutrition, Ministry of Agriculture Strategic Research Chair Program (SRP, University of Saskatchewan) was applied in this study. The fractionation, predicted biodegradation and biodigestion were evaluated using the Cornell Net Carbohydrate Protein System. The results show that: (1) The steam flaking induced significant changes in CHO subfractions, CHO biodegradation and biodigestion in ruminant system. There were significant differences between non-processed (raw) and steam flaked grain corn (P<.01); (2) The ATR-Ft/VMS molecular technique was able to detect the processing induced CHO molecular structure changes; (3) Induced CHO molecular structure spectral features are significantly correlated (P<.05) to CHO subfractions, CHO biodegradation and biodigestion and could be applied to potentially predict CHO biodegradation (R2=0.87, RSD=0.74, P<.01) and intestinal digestible undegraded CHO (R2=0.87, RSD=0.24, P<.01). In summary, the processing induced molecular CHO structure changes in grain corn could be revealed by the ATR-Ft/VMS vibrational molecular spectroscopy. These molecular structure changes in grain were potentially associated with CHO biodegradation and biodigestion.

Alteration of biomacromolecule in corn by steam flaking in relation to biodegradation kinetics in ruminant, revealed with vibrational molecular spectroscopy

Large scale of steam flaked corn has been used in dairy ration to maintain high milk production level. This study aimed to determine effects of steam flaking on processing-induced intrinsic molecular structure changes that were associated with rumen degradation kinetics and nutrients supply. The advanced vibrational molecular spectroscopy was applied to reveal the processing-induced intrinsic structure changes on a molecular basis. The rumen degradation kinetics and nutrient supply were determined using in situ approach in ruminant livestock system. Raw corn grain (RC) and steam flaked corn grain (SFC) were obtained from two different processing plants. The results showed that (1) Compared to RC, SFC had greater truly digestible non-fiber carbohydrate [tdNFC: 86.8 versus 78.0% dry matter (DM)], but lower truly digestible crude protein [tdCP: 7.7 versus 9.0% DM]. (2) The steam flaking increased (P<0.01) rumen degradable DM (RDDM) and starch (RDSt), but decreased (P<0.01) rumen degradable protein (RDP). (3) Molecular absorbance intensities of most carbohydrate biopolymers were greater in SFC (P<0.01), but protein amides associated molecular spectral intensities were lower (P<0.01) in SFC. (4). The molecular structure and nutrient interactive study showed that carbohydrate spectral intensities were positively (P<0.10) associated with RDDM and RDSt and protein amide spectral intensities were positively (P<0.10) associated with RDP. This results indicated that the steam flaking induced molecular structure changes had an interactive relationship with rumen degradation kinetics.

Physiochemical Characteristics and Molecular Structures for Digestible Carbohydrates of Silages.

The main objectives of this study were (1) to assess the magnitude of differences among new barley silage varieties (BS) selected for varying rates of in vitro neutral detergent fiber (NDF) digestibility (ivNDFD; Cowboy BS with higher ivNDFD, Copeland BS with intermediate ivNDFD, and Xena BS with lower ivNDFD) with regard to their carbohydrate (CHO) molecular makeup, CHO chemical fractions, and rumen degradability in dairy cows in comparison with a new corn silage hybrid (Pioneer 7213R) and (2) to quantify the strength and pattern of association between the molecular structures and digestibility of carbohydrates. The carbohydrate-related molecular structure spectral data was measured using advanced vibrational molecular spectroscopy (FT/IR). In comparison to BS, corn silage showed a significantly (P < 0.05) higher level of starch and energy content and higher degradation of dry matter (DM). Cowboy BS had lower feeding value (higher indigestible fiber content and lower starch content) and lower DM degradation in the rumen compared to other BS varieties (P < 0.05). The spectral intensities of carbohydrates were significantly (P < 0.05) correlated with digestible carbohydrate content of the silages. In conclusion, the univariate approach with only one-factor consideration (ivNDFD) might not be a satisfactory method for evaluating and ranking BS quality. FT/IR molecular spectroscopy can be used to evaluate silage quality rapidly, particularly the digestible fiber content.

Structural changes on a molecular basis of canola meal by conditioning temperature and time during pelleting process in relation to physiochemical (energy and protein) properties relevant to ruminants.

The objectives of this study were: (1) To investigate the effects of conditioning temperature (70, 80, 90°C), time (30, 60 sec), and interaction (temperature × time) during the pelleting process on internal protein molecular structure changes of the co-products; (2) To identify differences in protein molecular structures among pellets that were processed under different conditions, and between unprocessed mash and pellets; 3) To quantify protein molecular structure changes in relation to predicted energy and protein utilization in dairy cows. The final goal of this program was to show how processing conditions changed internal feed structure on a molecular basis and how molecular structure changes induced by feed processing affected feed milk value in dairy cows. The hypothesis in this study was that processing-induced protein inherent structure changes affected energy and protein availability in dairy cattle and the sensitivity and response of protein internal structure to the different pelleting process conditions could be detected by advanced molecular spectroscopy. The protein molecular structures, amides I and II, amide I to II ratios, α-helix structure, β-sheet structure, and α to β structure ratios, were determined using the advanced vibrational molecular spectroscopy (ATR-FT/IR). The energy values were determined using NRC2001 summary approach in terms of total digestible nutrients, metabolizable and net energy for lactation. The protein and carbohydrate subfactions that are related to rumen degradation characteristics and rumen undegraded protein supply were determined using updated CNCPS system. The experiment design was a RCBD and the treatment design was a 3x2 factorial design. The results showed that pelleting induced changes in protein molecular structure. The sensitivity and response of protein inherent structure to the pelleting depended on the conditioning temperature and time. The protein molecular structure changes were correlated (P < 0.05) with energy values and protein subfractions of the pelleted co-product. The results indicated that the protein internal molecular structure had significant roles in determining energy and protein nutritive values in dairy cows. Multi-regression study with model variables selection showed that the energy and protein profiles in pelleted co-products could be predicted with the protein molecular structure profiles. This approach provides us a relatively new way to estimate protein value in dairy cows based on internal protein molecular structure profile.