Effect of nitrogen fertilisation on the amino acid digestibility of different triticale genotypes in caecectomised laying hens.

BACKGROUND: The influence of nitrogen fertilisation and genotype on the amino acid (AA) digestibility of triticale grain was investigated in caecectomised laying hens. Three genotypes, Grenado, EAW6002 and Lasko, were cultivated with and without nitrogen fertilisation at the end of the heading stage. The six triticale variants as well as a basal diet were each used to feed seven laying hens in a 7 × 7 Latin square design. RESULTS: Nitrogen fertilisation influenced the digestibility of Cys, Glu, Phe and Ser in some triticale genotypes and reduced Ala, Ile, Lys, Met and Val digestibility in all genotypes (P < 0.05). Nitrogen fertilisation increased the concentration of all AAs in the grain. Consequently, the concentration of digestible AAs in the grains was increased for most AAs upon nitrogen fertilisation. Overall, Lys had the lowest digestibility, whereas that of Glu and Pro was the highest. For the triticale genotypes, the level of AA digestibility was highest for EAW6002 followed by Lasko and Grenado, with significant differences (P < 0.05) between genotypes for some but not all AAs. CONCLUSION: The results indicated that the accuracy of the digestible AA supply for hen feeding might benefit from considering fertilisation and genotype-specific digestibility data in feed formulation. © 2016 Society of Chemical Industry.

Changes in Rumen Microbial Community Composition during Adaption to an In Vitro System and the Impact of Different Forages.

This study examined ruminal microbial community composition alterations during initial adaption to and following incubation in a rumen simulation system (Rusitec) using grass or corn silage as substrates. Samples were collected from fermenter liquids at 0, 2, 4, 12, 24, and 48 h and from feed residues at 0, 24, and 48 h after initiation of incubation (period 1) and on day 13 (period 2). Microbial DNA was extracted and real-time qPCR was used to quantify differences in the abundance of protozoa, methanogens, total bacteria, Fibrobacter succinogenes, Ruminococcus albus, Ruminobacter amylophilus, Prevotella bryantii, Selenomonas ruminantium, and Clostridium aminophilum. We found that forage source and sampling time significantly influenced the ruminal microbial community. The gene copy numbers of most microbial species (except C. aminophilum) decreased in period 1; however, adaption continued through period 2 for several species. The addition of fresh substrate in period 2 led to increasing copy numbers of all microbial species during the first 2-4 h in the fermenter liquid except protozoa, which showed a postprandial decrease. Corn silage enhanced the growth of R. amylophilus and F. succinogenes, and grass silage enhanced R. albus, P. bryantii, and C. aminophilum. No effect of forage source was detected on total bacteria, protozoa, S. ruminantium, or methanogens or on total gas production, although grass silage enhanced methane production. This study showed that the Rusitec provides a stable system after an adaption phase that should last longer than 48 h, and that the forage source influenced several microbial species.

Studies on potential effects of fumaric acid on rumen microbial fermentation, methane production and microbial community.

The greenhouse gas methane (CH4) contributes substantially to global climate change. As a potential approach to decrease ruminal methanogenesis, the effects of different dosages of fumaric acid (FA) on ruminal microbial metabolism and on the microbial community (archaea, bacteria) were studied using a rumen simulation technique (RUSITEC). FA acts as alternative hydrogen acceptor diverting 2H from methanogenesis of archaea towards propionate formation of bacteria. Three identical trials were conducted with 12 fermentation vessels over a period of 14 days. In each trial, four fermentation vessels were assigned to one of the three treatment groups differing in FA dosage: low fumaric acid (LFA), high fumaric acid (HFA) and without FA (control). FA was continuously infused with the buffer. Grass silage and concentrate served as substrate. FA led to decreases in pH and to higher production rates of total short chain fatty acids (SCFA) mediated by increases in propionate for LFA of 1.69 mmol d(-1) and in propionate and acetate production for HFA of 4.49 and 1.10 mmol d(-1), respectively. Concentrations of NH3-N, microbial crude protein synthesis, their efficiency, degradation of crude nutrients and detergent fibre fraction were unchanged. Total gas and CH4 production were not affected by FA. Effects of FA on structure of microbial community by means of single strand conformation polymorphism (SSCP) analyses could not be detected. Given the observed increase in propionate production and the unaffected CH4 production it can be supposed that the availability of reduction equivalents like 2H was not limited by the addition of FA in this study. It has to be concluded from the present study that the application of FA is not an appropriate approach to decrease the ruminal CH4 production.

Effect of monensin on in vitro fermentation of silages and microbial protein synthesis.

The objective of the study was to investigate the effects of monensin on silage fermentation and microbial net protein synthesis. In Experiment 1, monensin (0.5, 1, 2, 4, 6, or 10 µg) was added to syringes that contained 120 mg of grass silage (GS), grass silage and concentrate (GS + C), or maize silage (MS), resulting in concentrations of 4.2, 8.3, 16.7, 33.3, 50.0 and 83.3 mg monensin/kg feed. Samples were incubated for 24 h to determine the monensin concentration that resulted in the maximum reduction in methane production without effects on the total gas production. In Experiment 2, GS and GS + C were incubated in a rumen simulation technique (Rusitec) to assess the monensin effects (133 and 266 mg/kg feed) on the production of total gas, methane and volatile fatty acids (VFA), degradation of nutrients and microbial net protein synthesis. In Experiment 1, methane production was reduced without significant effects on the total gas production; the reductions were 17% (GS), 10% (GS + C) and 13% (MS) with 16.7 (GS), 50.0 (GS + C) and 33.3 (MS) mg monensin/kg feed. Monensin reduced the total gas and methane production in GS and GS + C in Experiment 2. Propionate production was enhanced by monensin, accompanied by a decrease in acetate production. Along with a reduction in crude protein (CP) degradation, monensin reduced the ammonia nitrogen concentration in the effluent of both treatments. While the protein produced by liquid-associated microbes increased with monensin, protein production by solid-associated microbes was reduced. Total microbial net protein synthesis increased in the presence of monensin. Monensin influenced the production of total gas, methane and VFA from the silages without an effect on the degradation of organic matter (OM). Different microbial fractions were affected differently by monensin supplementation. If monensin is used as a tool to reduce methane emission, the supplementation level must be carefully chosen to avoid negative effects on overall fermentation in the rumen.

Influence of apple and citrus pectins, processed mango peels, a phenolic mango peel extract, and gallic Acid as potential feed supplements on in vitro total gas production and rumen methanogenesis.

Several food processing byproducts were assessed as potential feed and feed supplements. Since their chemical composition revealed a high nutritional potential for ruminants, the Hohenheim in vitro gas test was used to investigate total gas, methane, and volatile fatty acid production as well as protozoal numbers after ruminal digestion of different substrate levels. Processing byproducts used were low- and high-esterified citrus and apple pectins, integral mango peels, and depectinized mango peels. In addition, the effect of a phenolic mango peel extract and pure gallic acid was investigated. The highest decrease in methane production (19%) was achieved by supplementing high levels of low-esterified citrus pectin to the hay-based diet. Interestingly, total gas production was not affected at the same time. Showing valuable nutritional potential, all byproducts exhibited, e.g., high metabolizable energy (11.9-12.8 MJ/kg DM). In conclusion, all byproducts, particularly low-esterified citrus pectin, revealed promising potential as feed and feed supplements.

Investigations on the effect of forage source, grinding, and urea supplementation on ruminal fermentation and microbial protein flow in a semi-continuous rumen simulation system.

The objective of the present study was to compare the effect of maize silage and grass silage on microbial fermentation and protein flow in a semi-continuous rumen simulation system (Rusitec) when milling screen size (MSS) during grinding was varied. Oven-dried silages were milled through screens of 1, 4 or 9 mm pore size and incubated for 48 h in a Rusitec system. Furthermore, the effect of N supplementation to maize silage (MSS: 4 mm) was investigated and single dose vs. continuous infusion of urea-N were compared. Degradation of organic matter (OM), crude protein (CP), fibre fractions and non-structural carbohydrates (NSC) as well as short-chain fatty acid production differed significantly between forage sources. Urea-N supplementation improved the degradation of NSC, but not that of fibre fractions in maize silage. The way of urea supply had only marginal effects on fermentation characteristics. An increase in MSS, and consequently in mean feed particle size, led to an improvement in the degradation of OM, CP and NSC, but efficiency of microbial net protein synthesis (EMPS; mg microbial N flow/g degraded OM) and the microbial amino acid profile were less affected. EMPS was higher in grass silage than in maize silage and was improved by urea-N supplementation in maize silage. This study indicates that fermentation of NSC as well as EMPS during incubation of maize silage was limited by availability of NH3-N. Furthermore, an increase in MSS above 1 mm seems to improve fermentation of silages in the Rusitec system.

Effects of concentrate proportion in the diet with or without Fusarium toxin-contaminated triticale on ruminal fermentation and the structural diversity of rumen microbial communities in vitro.

The objective of this study was to investigate the effects of the concentrate proportion and Fusarium toxin-contaminated triticale (FCT) in the diet on nutrient degradation, microbial protein synthesis and structure of the microbial community, utilising a rumen simulation technique and single-strand conformation polymorphism (SSCP) profiles based on PCR-amplified small subunit ribosomal RNA genes. Four diets containing 60% or 30% concentrates on a dry matter basis with or without FCT were incubated. The fermentation of nutrients and microbial protein synthesis was measured. On the last day of incubation, microbial mass was obtained from the vessel liquid, DNA was extracted and PCR-primers targeting archaea, fibrobacter, clostridia, bifidobacteria, bacillii, fungi, and bacteria were applied to separately study the individual taxonomic groups with SSCP. The concentrate proportion affected the fermentation and the microbial community, but not the efficiency of microbial protein synthesis. Neither the fermentation of organic matter nor the synthesis and composition of microbial protein was affected by FCT. The fermentation of detergent fibre fractions was lower in diets containing FCT compared to diets with uncontaminated triticale. Except for the clostridia group, none of the microbial groups were affected by presence of FCT. In conclusion, our results give no indication that the supplementation of FCT up to a deoxynivalenol concentration in the diet of 5 mg per kg dry matter affects the fermentation of organic matter and microbial protein synthesis. These findings are independent of the concentrate level in the diets. A change in the microbial community composition of the genus Clostridia may be the reason for a reduction in the cellulolytic activity.

Determination of titanium dioxide supplements in different matrices using two methods involving photometer and inductively coupled plasma optical emission spectrometer measurements.

Titanium dioxide (TiO2) is a viable marker in digestibility studies using different animal species. The photometrical analysis is based on an intense orange colour following the addition of hydrogen peroxide to an acid solution. The measurement using inductively coupled plasma optical emission spectrometry (ICP-OES) is a method for analysing more than one element from the same sample preparation. The present study was conducted to investigate whether an established ICP-OES element analysis following acid-based hydrolysis is appropriate for titanium analysis. Defined amounts of TiO2 were added to samples obtained in studies with cows, pigs and turkeys and recoveries were determined. It was shown that supplemented TiO2 can be determined in samples of feeds, faeces or excreta, and digesta using a method based on photometric or ICP-OES measurement. The differences between the true and measured titanium concentrations indicate that using the ICP-OES method leads to a higher accuracy of determination.

Ruminal fermentation patterns and parameters of the acid base metabolism in the urine as influenced by the proportion of concentrate in the ration of dairy cows with and without Fusarium toxin-contaminated triticale.

Feeding a total mixed ration with 50% concentrate and a mean deoxynivalenol (DON) concentration of 5.3 mg/kg DM to 13 German Holstein cows in early lactation (Myco group) resulted in alterations in the ruminal fermentation patterns (lower molar percentage of acetate and isobutyrate, higher molar percentage of valerate) compared to the 14 control cows (Period 1, 11 weeks). In the Myco group, significantly lower ruminal pH value occurred in weeks 4 and 8 and lower minimum pH values critical for developing subacute ruminal acidosis were detected. Accordingly, the net acid base excretion in the urine and the base-to-acid ratio were lower (significant in week 8 only). These effects probably resulted from a higher dry matter intake and are not related to the presence of Fusarium toxin. In Period 2, the same 27 cows plus five additional cows were divided into four groups over 18 weeks. The effects of an elevated concentrate proportion of 60% were tested with and without Fusarium toxin (Control-60, 0.4 mg DON/kg DM and Myco-60, 4.6 mg DON/kg DM) and compared to two groups fed 30% concentrate (Control-30, 0.6 mg DON/kg DM and Myco-30, 4.4 mg DON/kg DM). As expected, a high concentrate proportion significantly affected the ruminal fermentation patterns. Net acid base excretion and base-to-acid ratio did not reveal a distinct concentrate effect and the ruminal pH values were on a high level in all groups when ruminal fluids were taken after 3-5 hours of feed restriction (median between 6.8 and 7.2). Additional effects were observed on the profile of short chain fatty acids in the presence of Fusarium toxin at both concentrate levels. This indicates a switch in the microbial community due to direct mycotoxin effects and/or indirect effects of the Fusarium infection related alterations in the physico-chemical properties of the infected cereal on ruminal microbes.

On the effects of the concentrate proportion of dairy cow rations in the presence and absence of a fusarium toxin-contaminated triticale on cow performance.

The aim of the present study was to investigate the effects of a deoxynivalenol (DON) contaminated ration with a concentrate proportion of 50%, on the performance of dairy cows (Period 1), and to examine the effects when the concentrate proportion was elevated to 60% compared to a ration with 30% concentrates (Period 2). In Period 1, 13 lactating German Holstein cows (Myco group, on average 29 days in milk) were fed the experimental diet (on average 5.3 mg DON/kg DM) as total mixed ration over 11 weeks, while another 14 cows (on average 33 days in milk) received a control diet. Both rations contained 50% concentrates (on DM basis). In Period 2 (18 weeks), the same 27 cows plus five additional cows were divided into four groups: Control-30 (30% concentrates), Myco-30 (30% concentrates, 4.4 mg DON/kg DM), Control-60 (60% concentrates), Myco-60 (60% concentrates, 4.6 mg DON/kg DM). The overall performance level was characterised by a mean daily DM intake of 17.9 kg and a mean daily milk production of 26.7 kg fat corrected milk (FCM) in Period 1 and 17.3 kg DM intake and 24.5 kg FCM in Period 2, respectively. In both periods cows fed the Fusarium toxin-contaminated diets consumed more DM (in Period 2 only significant for group Myco-30) resulting from stimulating effects on the ingesta passage rate of the natural contaminated Fusarium-infected triticale. In Period 1, cows fed the Fusarium toxin-contaminated diet had a significantly higher milk yield, milk urea and somatic cell count, whereas milk fat and protein concentration and fat-to-protein-ratio (FPR) were significantly lower. In Period 2, on a low concentrate level, FCM was significantly higher in group Myco-30. On a high concentrate level, group Myco-60 produced significantly more milk, but milk fat and protein concentration, FPR and milk urea were significantly lower. A concentrate proportion of 60% had a depressing effect on milk fat concentration but was significantly more pronounced in the presence of Fusarium toxin-contaminated and Fusarium damaged-triticale.

Description of the structural diversity of rumen microbial communities in vitro using single-strand conformation polymorphism profiles.

Changes of the rumen microbial community structure, as it can be established with a rumen simulation technique (RUSITEC) were studied using PCR and single-strand conformation polymorphism (SSCP) of small subunit rDNA genes (SSU rDNA). Four total mixed rations were incubated and two ammonia levels in the artificial saliva were applied. Three replicated vessels were used for each treatment. Mixed microbial fractions were isolated by stepwise centrifugation from the liquid fraction (reference microbes, RM) and from the solids of the feed residues (solid-associated microbes, SAM). PCR-primers targeting archaea, fibrobacter, clostridia, and bacteria, respectively, were applied to represent the individual taxonomic groups by SSCP profiles. These SSCP profiles were converted into a binary matrix and distances among treatments were visualised by non-metric multidimensional scaling. Between replicates belonging to one treatment only small differences were found, indicating a high reproducibility of the RUSITEC and the chosen SSCP method. The ammonia concentration seems to be affecting the SSCP profiles. Great differences occurred between RM and SAM, especially for profiles targeting bacteria and clostridia. Differences in the profiles of RM were also found between mixed rations that contained the same feedstuffs in different ratios and between rations with similar nutrient content but based on different feedstuffs. In conclusion, the PCR-SSCP-based technique in conjunction with non-metric multidimensional scaling was sufficiently sensitive to detect and compare changes in composition of rumen microbial community structure in vitro as affected by diet and other environmental factors.

Nutrient digestibility and prediction of metabolizable energy in total mixed rations for ruminants.

The aim of the present study was to determine equations that predict ME in total mixed rations (TMR) based on routine methods. The ME content of 30 TMR for dairy cows was determined based on digestible crude nutrients obtained with wether sheep. Concentrations in the TMR (in g/kg DM) varied between 118 and 234 for crude protein, 26 and 48 for crude lipid, 131 and 250 for crude fibre, 281 and 488 for NDF, and 173 and 304 for ADF. Gas production ranged from 40.7 to 54.1 ml/200 mg DM, and enzymatically degraded organic matter from 652 to 800 g/kg DM. Digestibility [%] ranged from 68.6 to 84.0 for organic matter, from 55.6 to 84.3 for crude lipid, from 55.0 to 77.8 for crude fibre, from 57.6 to 77.0 for NDF and from 53.1 to 79.6 for ADF. ME ranged from 9.6 to 11.9 MJ/kg DM, and NEL from 5.7 to 7.4 MJ/kg DM. ME content was highly correlated with the concentration of both crude fibre and enzymatically degradable organic matter as well as with organic matter digestibility. A multiple regression equation based on crude fibre and crude lipid predicted ME with a reasonable goodness of fit (r2 = 0.81; s(y.x) = 2.4%). The inclusion of other nutrients, of neutral and acid detergent fibre, neither of gas production did improve the goodness of fit. The best prediction was achieved with inclusion of enzymatically degraded organic matter (r2 = 0.90; s(y.x) = 1.7%).