Carlos Alberto Rodríguez

Composition of bacteria harvested from the liquid and solid fractions of the rumen of sheep as influenced by feed intake.

A study was conducted to determine the effect of the feed intake on the chemical composition of bacteria associated with the solid (solid-associated bacteria; SAB) and liquid (liquid-associated bacteria; LAB) fractions of rumen digesta, the digestive passage kinetics and their relationships. Whole rumen contents were sampled after a period of continuous infusion of 15NH3 from four ruminally-cannulated wethers provided successively with a hay-concentrate diet (2 : 1 w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight 0.75. SAB had a higher content of organic matter and total lipids (P < 0.001) and a similar N content as compared with LAB. The concentration of purines and 15N was lower (P = 0.011 and P < 0.001 respectively) in SAB than LAB, whereas the opposite was observed for the concentration of amino acids (mg/g DM; P = 0.031). An increase in feed intake produced an increase in the N (P = 0.034) and purine (P = 0.066) concentrations in bacteria and a decrease (P = 0.033) in their amino acid concentrations. Significant increases of rumen outflow rates of liquid and particles were also observed with increased feed intake. Rates of rumen outflow showed positive and negative linear relationships (P < 0.001) with the purine : N ratio and the proportion of amino acid on total N of bacteria respectively. SAB contained significantly higher proportions of leucine, isoleucine, lysine and phenylalanine and lower proportions of alanine, methionine and valine than LAB. The increase in feed intake also induced significant changes in the amino acid profile of bacteria, increasing arginine and methionine and decreasing alanine and glycine proportions. Results show that the outflow rate of rumen contents is a major factor in determining the proportion of nucleic acids and protein in rumen bacteria and explains some of the differences observed between LAB and SAB.

In situ study of the relevance of bacterial adherence to feed particles for the contamination and accuracy of rumen degradability estimates for feeds of vegetable origin

An in situ study was conducted on four rumen-cannulated wethers to determine (using (15)N infusion techniques) the microbial contamination (mg bacterial DM or crude protein (CP)/100 mg DM or CP) and the associated error on the effective degradability of fourteen feeds: barley and maize grains, soyabean and sunflower meals, full-fat soyabean, maize gluten feed, soyabean hulls, brewers dried grains, sugarbeet pulp, wheat bran, lucerne and vetch-oat hays, and barley and lentil straws. The DM or CP contamination in residues (M) fitted to single exponential or sigmoid curves. A general model (M=m (1-e(-ft) ) (j)) was proposed to match this fit. Asymptotic values (m) varied from 2.84% to 13.3% and from 2.85% to 80.9% for DM and CP, respectively. Uncorrected results underestimated the effective degradability of both DM (P<0.05) and CP (P<0.01). For CP, this underestimation varied from 0.59 % to 13.1%, with a higher but unascertainable error for barley straw. Excluding maize grain, the microbial contamination of both DM and CP, and the associated underestimation of the effective degradability of CP, were positively related to the cellulose content of the feed. The error in the effective degradability of CP was also negatively related to the CP content and its apparent effective degradability (R(2) 0.867). This equation allows easier and more accurate estimates of effective degradability, needed to improve protein-rationing systems.

Effect of diet forage to concentrate ratio on rumen degradability and post-ruminal availability of protein from fresh and dried lucerne

The ruminal degradation of dry matter (DM) and crude protein (CP) and the intestinal availability of CP of four lucerne samples were measured on two diets with lucerne hay to concentrate ratios of 2: 1 (diet F) and 1: 2 (diet C). Two samples of fresh lucerne (third cut) harvested after 2 (FL 1 ) or 8 (FL 2 ) weeks from the previous cut were used together with a sample of lucerne hay (LH) and another of dehydrated lucerne (DL). Rumen degradability, measured by the nylon bag technique, and rumen outflow rates were determined on three rumen cannulated wethers. Intestinal digestibility was determined by the mobile bag technique on three duodenal fistulated wethers. For CP, significantly lower values were observed with diet C than with diet F for the potentially degradable insoluble fraction (0·334 v. 0·397) and its degradation rate (0·093 v. 0·134 per h). As a consequence, the effective degradability was also lower with diet C (0·746 v. 0·821; P = 0·059). Effective degradability of DM was also apparently lower with diet C (0·596 v. 0·634). With both diets, the intestinal digestibility decreased in all the samples with increase of ruminal incubation time according to a simple exponential equation. The undegraded CP digested in the gut (Di) and therefore the effective intestinal digestibility (EID) were derived from this exponential function according to the rumen outflow of undegraded CP. Mean values of Di (expressed as proportion of food CP content) were respectively 0·091 and 0·142 for F and C diets and 0·084, 0·115, 0·116, and 0·152 for FL 1 , FL 2 , LH and DL samples. Lower rumen degradability was partially compensated for by higher Di values resulting in a close correlation between both parameters (r = –0·965; P 0·001). The change of the digestion site associated with the reduction of the effective degradability of CP produced also an increase in the undigested CP as a proportion of food CP. So, these values are respectively 0·087 and 0·112 for F and C diets and 0·053, 0·109, 0·096, and 0·141 for FL 1 , FL 2 , LH, and DL samples. No difference in EID between F and C diets was observed (0·529 v. 0·563). For samples, the only effect (P 0·05) was recorded between FL 1 (0·618) and the other samples (0·509, 0·544 and 0·512 for FL 2 , LH, and DL, respectively).

Effects of feed intake on composition of sheep rumen contents and their microbial population size

The present study was conducted to determine the effect of feed intake on the composition of the rumen contents of sheep and on their bacterial densities. Whole rumen contents were sampled after a period of continuous inter-rumen infusion of 15NH3 from four rumen-cannulated wethers successively fed on a hay-concentrate diet (2:1, w/w on a DM basis) at two rates of feed intake: 40 and 80 g DM/kg body weight0.75. Total weight and chemical composition of rumen contents, as well as the distribution by size and chemical composition of particles, were determined. The populations of bacteria associated with the liquid (liquid-associated bacteria, LAB) and solid (solid-associated bacteria, SAB) fractions of rumen digesta and the distribution of SAB according to feed particle size were also examined. The greater feed intake caused an increase in the mass of the rumen contents, while its chemical composition did not change, except for a higher content of organic matter (P=0.023). The distribution of feed particles by size was similar at both levels of intake. The concentrations of neutral- and acid-detergent fibre in feed particles decreased and those of total, dietary, and microbial N increased, both with a quadratic response (P=0.001), as particle size decreased. The proportion of LAB in the microbial biomass of rumen digesta reached only 8.0 %. This proportion and the density of LAB were unaffected by the level of feed intake, whereas an apparent reduction (10.4 %) occurred with the SAB biomass in whole rumen contents. A systematic, but not significant, reduction (mean value 11.9 %) in the level of microbial colonisation in the different particle fractions with the increase of feed intake was also observed.

Composition of free and adherent ruminal bacteria: inaccuracy of the microbial nutrient supply estimates obtained using free bacteria as reference samples and 15 N as the marker

Previous studies have indicated that (15)N enrichment of solid-associated bacteria (SAB) may be predicted from the same value in liquid-associated bacteria (LAB). The aims of this study were to confirm this and to measure the error in the nutrient supply from SAB, when LAB are used as the reference sample. For this purpose, the chemical and amino acid (AA) compositions of both the bacterial populations were studied in four experiments carried out on different groups of three rumen cannulated wethers. Diets (one in Experiments 1 and 4 and three in Experiments 2 and 3) had forage-to-concentrate ratios (dry matter (DM) basis) between 2 : 1 and 40 : 60, and were consumed at intake levels between 40 and 75 g DM/kg (BW)(0.75). The bacteria samples were isolated after continuous infusion of ((15)NH(4))(2)SO(4) (40, 18, 30 and 25 mg (15)N/day, in Experiments 1 to 4, respectively) for at least 14 days. In all experiments, SAB had consistently higher concentrations of organic matter (826 v. 716 g/kg DM, as average) and total lipids (192 v. 95 g/kg DM, as average) than LAB. Similar CP concentrations of both populations were observed, except a higher concentration in SAB than in LAB in Experiment 3. A consistent (in Experiment 4 only as tendency) higher AA-N/total N ratio (on average 17.5%) was observed in SAB than in LAB. The (15)N enrichment in SAB was systematically lower than in LAB. On the basis of the results of all studies a close relationship was found between the (15)N enrichment in SAB and LAB, which was shown irrespective of experiments. This relationship was established from Experiments 1 and 2 and the above cited previous results (n = 20; P < 0.001; R(2) = 0.996), and then confirmed from the results of Experiments 3 and 4. These relationships between SAB and LAB demonstrate that CP supply from SAB is underevaluated by, on average, 21.2% when LAB are used as the reference. This underevaluation was higher for true protein and even higher for the lipid supply (32.5% and 59.6%, respectively, as an average of the four experiments). Large differences in AA profile were observed between SAB and LAB. The prediction equation obtained using (15)N as the marker may be used to correct the errors associated with the traditional use of LAB as the reference sample, and therefore to obtain more accurate estimates of the microbial nutrient supply to the ruminants.

A simplified management of the in situ evaluation of feedstuffs in ruminants: Application to the study of the digestive availability of protein and amino acids corrected for the ruminal microbial contamination

The ruminal effective degradability (RED) and intestinal effective digestibility (IED) for dry matter, crude protein (CP) and amino acids (AA) were estimated by a simplified in situ method using pooled samples from rumen-incubated residues, which represented the ruminal outflow of undegraded feed. The effect of microbial contamination in the rumen was corrected using 15N infusion techniques. Studies were carried out for soybean meal (SBM), barley grain (BG) and lucerne hay (LH) in three wethers cannulated in the rumen and the duodenum. Uncorrected values of RED for CP obtained either by mathematical integration or our simplified method were similar in all feeds. Microbial N in the pooled samples of SBM, BG and LH were 2%, 11% and 24% of total N, respectively. However, intestinal incubation eliminated this microbial charge by 100%, 99% and 88%, respectively. With microbial corrections, RED showed an increase, and IED showed a decrease, except for SBM. With this correction, intestinal digested CP was reduced by 2% in SBM, 13% in BG and 34% in LH. Corrected IED of AA was relatively similar in SBM (97–99%). However, large variations were observed in BG (74–93%) and in LH (10–88%). Digestion in the rumen and intestine changed the essential AA pattern. Overall, our results support that AA digestion is affected by the characteristics of their radicals and their contents in plant cell wall proteins. The accurate estimation of feed metabolisable AA or protein requires effective measures that are corrected by ruminal microbial contamination. The proposed in situ method largely simplifies these tasks and allows a more complete and less expensive feed evaluation.

Protein value of cereals and cereal by-products for ruminants: a comparison between crude protein and protein-based estimates.

In situ estimates of ruminal undegraded fraction (RU) and effective intestinal digestibility (EID, corrected for microbial colonisation) of dry matter (DM), crude protein (CP) and total analysed amino acids (TAA) of rye, wheat and corn grains, wheat bran, wheat and barley distillers’ dried grains with solubles (DDGS) and corn gluten feed were measured on three rumen and duodenum cannulated wethers using 15N labelling techniques and considering ruminal rates of particle comminution (kc) and outflow. Results indicate that not considering kc and microbial colonisation led to considerable overestimations of RU which increased with feed ruminal degradation. Microbial colonisation may be also associated with overestimations of EID, whose estimates for DM, CP and TAA were predicted from parameters related with the ruminal escape of intestinally indigestible materials. The RU estimates were higher for TAA than for CP in grains, but the opposite was observed in by-products, whereas EID estimates were higher for TAA in all feeds. To obtain accurate protein values in these feedstuffs, it is required to consider both kc and ruminal microbial colonisation. The CP-based results underestimate the intestinally digested protein in grains and the opposite is evidenced in cereal by-products. Microbial protein synthesised in the rumen is largely the major fraction of the feedstuff protein value with the exception of DDGS.

Understanding cloud dynamics using a ground-based radar at 94 GHz

This paper presents the analysis of mixed-phase cloud observations in Madrid (Spain) carried out with a 94-GHz ground-based CW-LFM radar system. This analysis is based on the 3D reflectivity profiles and the Doppler spectra. The case of interest occurred on 29th January 2016 when a melting layer at 1 Km, up- and downdrafts and drizzle were detected. This radar can be used to estimate the microphysical composition of the detected mixed-phase clouds.