Vicente Javier Moya

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.

Effects of the comminution rate and microbial contamination of particles in the rumen on in situ estimates of protein and amino acid digestion of expeller palm kernel and rapeseed meal

BACKGROUND Microbial corrected effective in situ estimates of ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of dry matter (DM), crude protein (CP) and amino acids (AA) of expeller palm kernel (EPK) and rapeseed meal (RSM) were measured on three rumen- and duodenum-cannulated wethers using ¹⁵N labelling techniques and considering ruminal rates of comminution (k(c)) and outflow (k(p)) of particles. RESULTS The lack of k(c) and microbial correction overestimated the RU of DM by 4.91% (EPK) and 9.88% (RSM). The lack of this correction also overestimated in both feeds the RU of CP, individual and total (TAA) AA as well as the IED of DM, CP, TAA and most AA. RU estimates were higher for CP than for TAA, but the opposite was observed for IED. The intestinal digested fraction was higher for CP than for TAA: 17.4% (EPK) and 13.8% (RSM). Digestion led to large changes in the essential AA profile in both feeds. CONCLUSION The lack of k(c) and microbial correction as well as CP-based results leads to considerable overestimations in the protein use of both feeds. Digestion aggravates the lysine deficiency of EPK but has global positive effects in the absorbed profile of RSM.

Effects of the correction of particle microbial contamination and particle transit model in the rumen on in situ protein evaluation of grass hays

Effects of considering the particle comminution rate (kc) in addition to particle rumen outflow (kp) and the ruminalmicrobialcontaminationonestimatesofby-passandintestinaldigestibilityofDM,organicmatterandcrudeprotein were examined in perennial ryegrass and oat hays. By-pass kc-kp-based values of amino acids were also determined. This studywasperformedusingparticletransit,insituand 15 Ntechniquesonthreerumenandduodenum-cannulatedwethers.The above estimates were determined using composite samples from rumen-incubated residues representative of feed by-pass. Considering thecomminutionrate,kc,modifiedthecontribution of theincubatedresidues tothesesamples inbothhays and revealed a higher microbial contamination, consistently in oat hay and only as a tendency for crude protein in ryegrass hay. Notconsideringkcorrumenmicrobialcontaminationovervaluedby-passandintestinaldigestibilityinbothhays.Therefore, non-microbial-correctedkp-basedvaluesofintestinaldigestedcrudeproteinwereoverestimatedascomparedwithcorrected andkc-kp-basedvalues inryegrass hay(17.4vs4.40%) andinoathay(5.73vs0.19%).Both factors shouldbeconsideredto obtain accurate in situ estimates in grasses, as the protein value of grasses is very conditioned by the microbial synthesis derived from their ruminal fermentation. Consistent overvaluations of amino acid by-pass due to not correcting microbial contamination were detected in both hays, with large variable errors among amino acids. A similar degradation pattern of amino acids was recorded in both hays. Cysteine, methionine, leucine and valine were the most degradation-resistant amino acids.

Determination of methane production from lactating goats fed diets with different starch levels

The methane (CH4) production was determined by indirect calorimetry. Twelve Murciano-Granadina goats at late lactation were fed with two isoenergetic diets (17.46 MJ/kg DM) with two different quantities of starch (29.03% vs. 14.44%) in a cross over design. Alfalfa hay and barley straw were the source of forage and mixes of concentrate (65%) were used to achieve the requirements during lactation. The goats were allocated to individual metabolism cages. After 8 d of adaptation, feed intake, total fecal and urine output, and milk yield were recorded daily during a 5 d period. Then, gas exchange measurements were recorded by a mobile open-circuit respirometry system using a face mask. Methane production was 60.60 and 100.37 kJ/kg BW0.75/d for the diet high and low in starch, respectively. Not significant differences were observed for milk production and composition between diets.The methane conversion factor was higher for diet lowest in starch (6.70% vs. 3.26%).

Interferences on microbial inhibitor tests related to ivermectin treatment in lactating dairy goats.

This Research Communication reports interferences related to the administration of ivermectin in lactating dairy goats on the response of microbial tests for screening antibiotics in milk. Twenty-eight Murciano-Granadina goats, naturally infested with Sarcoptes scabiei var. caprae, were treated with a subcutaneous injection of ivermectin (200 µg/kg b.w.). To prevent re-infestation, a second dose was applied 7 d later. Individual milk samples were collected, daily, up to 15 d post-treatment. Milk samples were analysed by microbial inhibitor tests (BRT MRL, Delvotest SP-NT MCS and Eclipse 100) and ivermectin residues were quantified by HPLC. A large number of positive results were obtained for all microbial tests, especially on the first day after treatment (BRT MRL = 46·4%; Delvotest SP-NT MCS = 14·3%; and Eclipse 100 = 17·8%). However, the highest concentration of drug residues in milk (24·3 ng/ml) was detected on the tenth day after treatment, when positive outcomes were relatively lower (BRT MRL = 17·8%; Delvotest SP-NT MCS = 10·7%; and Eclipse 100 = 7·4%). Results herein suggest that factors related to the ivermectin treatment other than drug residues in milk, or alterations produced by the parasitic disease itself affecting the immune response of animals, could be the cause of false-positive results in microbial tests. It can be concluded that the application of ivermectin in dairy goats infested with sarcoptes mange during lactation produces persistent drug residues in milk, and could also cause false-positive results in microbial inhibitor tests for screening antibiotics.

Amino acid availability in ruminants of cereals and cereal co‐products

BACKGROUND Microbial corrected in situ estimates of the ruminal undegraded fraction (RU) and intestinal effective digestibility (IED) of amino acids (AA), except tryptophan, of rye, wheat and corn grains, wheat bran, wheat and barley distilled dried grains and corn gluten feed were measured on three rumen- and duodenum-cannulated wethers using (15)N-labelling techniques and considering ruminal rates of particle comminution and outflow. RESULTS The lack of microbial correction led to overestimations of the intestinal digested fraction that rose with the increase in ruminal degradability. Thus these overestimations varied widely among feeds (from 4.3 to 32.1% for total analysed AA) and among AA. Digestion led to large changes in the AA supply that were greater in the rumen than in the intestine. The impact of these changes on the protein value is conditioned by the magnitude of the undegraded protein fraction. CONCLUSION Microbial contamination taking place in the rumen and changes in the AA supply with digestion should be considered to attain accurate estimates of AA digestion. Globally, digestion improved the AA supply in rye, wheat and wheat distilled dried grain and decreased it in corn and corn gluten feed by reducing the supply of valine and basic AA, especially lysine.