A systematic study of sheep faeces to be used as an alternative inoculum source in the Hohenheim gas test.

In vitro studies of gas production (GP) using rumen fluid obtained from rumen-cannulated animals are common in feed evaluation for ruminants. The objective was to compare sheep faeces with rumen fluid from dairy cows as inocula in the Hohenheim gas test (HGT) using a large number of different-quality feeds. It then was evaluated whether GP obtained by using rumen inoculum (RI-GP) can be estimated from GP using faecal inoculum (FI-GP). Ninety feeds for ruminants including roughages (n = 27), total mixed rations (n = 6), commercial compound feeds for dairy cows (n = 18), cereal grains (n = 10), energy-rich by-products (n = 5), legume grains (n = 6), oilseed meals (n = 6), and other protein-rich feeds (n = 12) were incubated with either rumen inoculum (RI) or faecal inoculum (FI), and GP was recorded at 2, 4, 8, 12, 24, 48, and 72 h. Nonlinear equations were fitted to the GP data to determine GP kinetic parameters. Correlations between RI-GP and FI-GP were calculated for different time points. Linear regressions were calculated to estimate RI-GP from FI-GP using the entire data set or subsets of roughages and concentrates. GP kinetic parameters were lower for all feed categories when incubated with FI instead of RI. On average of all feed samples, the potential GP was 9 mL/200 mg dry matter (DM) lower and the rate of GP was 3.1%/h lower with FI than RI. The estimation of kinetic data for FI included a lag phase of 1.51 h on average, whereas no lag phase was estimated for RI. Estimated parameters indicated an overall lower fermentation activity of FI than RI. However, there was a very similar progression of GP curves of RI-GP and FI-GP. RI-GP after 24 h was significantly correlated with FI-GP in the time period of 24 to 60 h (r = 0.973 - 0.982, including all feed samples). Linear regression analysis showed that RI-GP after 24 h can be estimated from FI-GP after 24 h (Slope = 1.02, R2 = 0.97) and 48 h (Slope = 1.1, R2 = 0.97) including the complete data set. Splitting the data set into roughages and concentrate feeds did not lead to distinctly higher estimation accuracy. In conclusion, sheep faeces can be used as an alternative inoculum for studying a wide range of different-quality feedstuffs. The standard gas test may be modified without the need for rumen-cannulated animals.

Assessing the combination efficiency of some unconventional feed resources with concentrates and Chloris gayana grass in mitigating ruminal methane production in vitro.

In a preliminary in vitro study, leaves of Acacia nilotica, Prosopis juliflora, Cajanus cajan, Leucaena leucocephala and seed kernel of Mangifera indica were identified as potential candidates in mitigating ruminal methane (CH4) production. The objective of the current study was to investigate the combination efficiency of these unconventional feeds with concentrate mix (CM) or Chloris gayana grass in CH4 reduction. Two feed combinations in different proportions were incubated in vitro with buffered rumen fluid at Hohenheim Gas test. In combination 1, C. gayana and CM were included as basal substrates, while in combination 2, A. nilotica, P. juliflora, C. cajan, L. leucocephala or M. indica seed kernel were included as CH4 reducing supplements at different proportions. The CH4 reducing potentials of feed combinations were presented as the ratio of CH4 to net gas production and expressed as percentage (pCH4). The pCH4 for CM and C. gayana was 16.7% and 16.9%, respectively, while it ranged from 3.18% in A. nilotica to 13.1% in C. cajan. The pCH4 was reduced (p < 0.05) from 14.6% to 9.39% when A. nilotica was combined with CM. In combination of L. leucocephala or C. cajan with CM, the pCH4 (p < 0.05) was reduced from 16.5% and 16.6% with the lowest proportion to 15.1% and 15.2% with the highest inclusion rate respectively. The combination of C. gayana with L. leucocephala or C. cajan reduced (p < 0.05) the pCH4 from 16.3% and 16.4% to 15.1% and 14.9% respectively. The pCH4 was reduced (p < 0.05) from 13.4% to 7.60% when A. nilotica was combined with C. gayana. Estimated digestible organic matter (dOM) and metabolizable energy (ME) increased (p < 0.05) with increasing proportions of M. indica seed kernel with CM or C. gayana. In conclusion, the combination of the basal substrates with unconventional supplements resulted in CH4 reduction without affecting the dOM and ME at lower inclusion rates. Animal-based experiments await to validate in vitro findings.

Microbial incubations of 8-phenyloctanoic acid and furan fatty acids in rumen fluid.

AIMS: The digestive tract of ruminants is specialized in the digestion of various plant components. One of the largest parts of the stomach is the so-called rumen, which contains a large number of micro-organisms that may degrade or modify fatty acids, for example by ß-oxidation, chain elongation and/or hydrogenation. METHODS AND RESULTS: Here we performed incubation experiments with less common fatty acids by in vitro incubations with rumen fluid of fistulated cows for 24 h. Sample extracts were analysed by gas chromatography with mass spectrometry. As substrates, we selected one phenyl fatty acid and four furan fatty acids (FuFAs). All studied fatty acids were degraded by ß-oxidation (two or three chain-shortening steps) while chain elongation or saturation of the aromatic part (terminal phenyl or central furan moiety) was not observed in any case. CONCLUSIONS: The percentage of ß-oxidation products was low, especially in the case of the FuFAs. This could be due to the rather long carbon number of FuFAs (19-22 carbon atoms). In addition, compound-specific differences in the degradation rates were observed in our experiments. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results produce evidence that FuFAs, which are valuable antioxidants that are known to be present in various feed items of the cow, can be effectively passed on the rumen into the milk.

In situ ruminal disappearance of crude protein and phytate from differently processed rapeseed meals in dairy cows.

BACKGROUND: The influence of different processing conditions of rapeseed meal on ruminal degradation of crude protein and phytate in dairy cows was investigated. Following oil extraction from the rapeseed, five residence times in the desolventizer/toaster were chosen to remove the solvent from the meal. Rapeseed cake and rapeseed meals were incubated in situ in the rumen of three fistulated dairy cows to determine ruminal degradation parameters. RESULTS: With increasing residence time in the desolventizer/toaster the ruminal degradation of crude protein decreased significantly for every treatment step. Ruminal phytate degradation and crude protein degradation were affected almost identically. CONCLUSION: The processing conditions of rapeseed meal have a major impact on the ruminal degradation of crude protein and phytate, indicating a potential conflict of interest regarding the production process. Large amounts of undegradable rumen protein are often intended for high-yielding dairy cows whereas a high level of ruminal degradation is preferred for phytate to increase absorption of phosphorus in the small intestine. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

In situ crude protein and starch degradation and in vitro evaluation of pea grains for ruminants.

Thirteen pea grain samples from different origins were used to examine the variation in in situ ruminal degradation of crude protein (CP) and starch as well as in vitro gas production (GP) kinetics. In vitro GP was used to estimate the digestibility of organic matter (dOM), metabolisable energy (ME) and utilisable CP at the duodenum (uCP). Protein fractions were also determined according to the Cornell Net Carbohydrate and Protein System. Degradation of CP and starch from all pea grains in the rumen was almost complete, with a high proportion of the instantly disappearing fraction. The variation in the degradation constants among pea grain variants was high, and degradation of CP and starch showed a significant initial lag phase in the rumen. The mean effective degradation (ED) calculated for a rumen outflow of k = 8%/h of CP (EDCP8) was higher than ED of starch (EDST8), averaging 77.0 and 71.5%, respectively, with low variation among variants. A correlation analysis between GP parameters and in situ degradation constants showed no clear relationship, but the rates of in vitro GP and in situ starch degradation were similar. Most of the protein in the pea grains was buffer-soluble with fast and intermediate degradation. Variation in the protein fractions among the pea grain variants was low and not suitable for predicting differences in in situ degradation characteristics. The mean in vitro uCP of pea grains was 198 g/kg dry matter (k = 8%/h) and variation was low and consistent with that of GP kinetics and in situ rumen undegradable crude protein values. The estimation of dOM and ME from 24 h GP led to very high values indicating that the existing prediction equations may not be suitable for pea grains as a single feed.

Ruminal degradation and estimated energy and protein values for ruminants of Durum wheat varieties grown in three locations.

Durum wheat is mainly used in pasta production, but may also be used as animal feed, for example as concentrate for dairy cows. Data on the ruminal degradation of Durum grains are scarce. Hence, the objective of the present study was to describe ruminal in situ crude protein (CP) and starch (ST) degradation, to investigate in vitro gas production kinetics, and to estimate utilisable CP at the duodenum (uCP) in vitro of five Durum varieties from three growing locations. Metabolisable energy (ME) and digestibility of organic matter (dOM) were also estimated using in vitro data. In situ incubations were conducted in three lactating jersey cows over defined timespans from 1 to 72 hr. Ruminal degradation parameters were estimated using exponential regression, and effective ruminal degradation was predicted for a ruminal passage rate of 8%/hr (ED8 ). In situ CP (a = 11%-19%; b = 80%-88%; c = 23%-33%/hr) and ST (a = 22%-39%; b = 60%-78%; c = 61%-123%/hr) degradation kinetics varied among samples and were influenced by location. Some samples showed a remarkably high ST degradation rate of up to 123%/h, which can significantly influence rumen pH and health when high amounts are incorporated into dairy rations. However, the ED of CP (77%-82%) and ST (91%-95%), and the in vitro estimates of ME (13.6-14.1 MJ/kg DM), dOM (92%-96%), and uCP (183-195 k/g DM) varied within a relatively small range. Hence, it may be adequate to use the mean values of these Durum grain characteristics when rations for ruminants are calculated and a differentiation depending on variety and location may not be necessary.

Determination of in situ ruminal crude protein and starch degradation values of compound feeds from single feeds.

Dairy cows are commonly fed compound feed concentrates, whose accurate formulation relies on the additivity of ruminal degradation characteristics of single feeds, and the absence of associative effects. The main aim of this study was to evaluate the additivity of single feeds in compound feeds made thereof. Twelve single feeds were used to produce eight compound feeds in mash and pelleted form. Samples of single and compound feeds were incubated in situ in three ruminally fistulated dairy cows, and effective ruminal degradation (ED) of CP and starch (ST) was computed. The ED values of examined compound feeds could be, in most cases, accurately calculated from ED values of single feeds. Observed EDCP values were significantly lower than that calculated, but differences were overall small and not exceeded 5% points. No significant differences were observed between calculated and observed EDST. The study also examined the effects of pelleting of compound feeds on in situ degradation. Pelleting significantly increased EDCP (up to 8% points), and EDST (up to 4% points) of most compound feeds. This could have been caused by the pelleting process increasing the proportion of fine feed particles with fast disappearance from the bags. It was concluded that small associative effects between the examined single feeds could be disregarded when formulating compound feeds for dairy cows, and that additivity of EDCP and EDST can be assumed in most cases.

Feed Clusters According to In Situ and In Vitro Ruminal Crude Protein Degradation.

Effective degradation (ED) of crude protein (CP) was estimated in vitro at 0.02, 0.05 and 0.08 h−1 assumed ruminal passage rates for a total of 40 feedstuffs, for which in situ ED was available and used as reference degradation values. For this, the Streptomyces griseus protease test was used. The differences between in vitro CP degradation and the in situ CP degradation values were lowest in legume grains and highest in cereal by-products and barley. The differences between in situ and in vitro ED were expressed using a degradation quotient (degQ), where degQ = (EDin vitro − EDin situ)/EDin situ. Among the tested feedstuffs, eight specific clusters were identified according to degQ for the assumed passage rates. The feedstuffs clustered in an unspecific way, i.e., feedstuffs of different nutrient composition, origin or treatment did not necessarily group together. Formaldehyde−treated rapeseed meal, soybean meal, wheat, a treated lupin, sunflower meal and barley could not be assigned to any of the clusters. Groupwise degradation (range of degQ for assumed passage rates are given in brackets) was detected in grass silages (−0.17, −0.11), cereal by-products together with sugar beet pulp (−0.47, −0.35) and partly in legume grains (−0.14, 0.14). The clustering probably based on different specific nutrient composition and matrix effects that influence the solubility of feed protein and limit the performance of the protease. The matrix can be affected by treatment (chemically, thermally or mechanically), changing the chemical and physical structure of the protein within the plant. The S. griseus protease test had reliable sensitivity to reflect differences between native feedstuffs and treatments (thermally or chemically) that were found in situ. The in situ results, however, are mostly underestimated. The clustering results do not allow a clear conclusion on the groupwise or feed-specific use of carbohydrate-degrading enzymes as pre- or co-inoculants as part of the S. griseus protease test and need to be tested for its potential to make this test more conform with in situ data.