Approaches to improve efficiency of N utilisation on dairy cow level

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1 Final REDNEX Conference, FIAP, Paris 30 August 2013 Approaches to improve efficiency of N utilisation on dairy cow level Mogens Vestergaard Aarhus University, Denmark

2 Why are dairy cows a concern? URINE N N INTAKE 503 g/day 37% Mills et al 2009 MILK N 28% FAECAL N 33% 2

3 Nitrogen in milk, faeces or urine, g/day Meta Analysis of N Balance in Dairy Cows Urine N Faecal N 200 Milk N Mills et al., Nitrogen intake, g/day 3

4 Mills et al., 2009 Milk N/N Intake vs. N Intake

5 Approaches to improve efficiency of N utilisation on dairy cow level Research on N digestion and N metabolism: Rumen (WP2) Amino acid and nutrient metabolism (WP3) N recycling (WP4)

6 WP2 Maximisation of absorption of feed and microbial protein in low-n diets Michel Doreau (WP leader), Pierre Nozière, Diego Morgavi Jamie Newbold, Jon Moorby Alejandro Belanche Sergio Calsamiglia, Alfred Ferret Andreas Foskolos Peter Lebzien Martina Aschemann

7 1 Innovative ways to decrease ruminal protein degradation Feed Crude Protein Degraded Protein energy minerals NH 3 rumen Undegraded Protein Microbial Protein small intestine Microbial Protein 2 Role of the microbial Ways to optimise ecosystem microbial protein especially protozoa synthesis 3

8 1. Innovative ways to decrease ruminal protein degradation State-of-the-art : the most efficient additives for decreasing ruminal protein degradation are various essential oils, but their effect in vivo remains unclear A first innovative approach was the inclusion of essential oils in silages. Essential oils in ryegrass silage are efficient in reducing silage protein degradation but the dose required and thus the cost may be too high for practical use A second innovative approach was the inclusion of polyclonal antibodies against proteolytic or deaminating bacteria. Several attempts failed to decrease protein degradation Capsicum oil and PTSO (propyl-propyl thiosulphate, derived from garlic oil processing) were promising additives. We observed a trend to an incrase in the ratio between N in milk and N intake with both additives (see next figure)

9 1. N Balance study with two essential oils (Capsicum oil and PTSO) Total N balance (g/day) N intake CTR 539 CAP 561 PTSO 505 Urine N CTR 216 CAP 219 PTSO 211 MNE (%) CTR 30.3 CAP 30.8 PTSO 34.3 N milk CTR 160 CAP 171 PTSO 175 Fecal N CTR 203 CAP 211 PTSO 188

10 2. Ways to optimise microbial protein synthesis In vitro, microbial protein yield did not differ between ryegrass and red clover, but microbes were more efficient with ryegrass for capturing N and in efficiency of N utilization in the rumen. Comparison of varieties (ryegrass varying in sugar content, red clover varying in polyphenol oxidase) needs further research Decreasing crude protein level to 11-12% of dietary DM in dairy cows resulted in a moderate decrease in milk production in 1 st experiment, but not in 2 nd experiment. In both experiments, urinary N was strongly decreased Decreasing crude protein level to 11-12% of dietary DM in dairy cows had a minor effect on the efficiency of microbial protein synthesis: no change in 1 st experiment, and only a trend to a decrease in 2 nd experiment The source of dietary carbohydrates, starch or fibre, had a minor effect on ruminal protein metabolism; a trend to a higher microbial protein flow and efficiency of synthesis was observed with starch (see next Table) Niacin supplementation to low-n diets did not change microbial protein flow and efficiency of synthesis but increased protozoa population; the use of fermentable protein by microbes may be changed (see next Table)

11 Normal N (14% CP) Low N (11% CP) Starch Fibre Starch Fibre OM digestibility, % N** Microbial N efficiency g N / kg OM fermented ns Fanchone et al., 2013 Urea (15.6% CP) Low (12% CP) Low +Niacin (12% CP) OM digestibility, % 72 a 69 b 71 ab * Microbial N efficiency g N / kg OM fermented ns Aschemann et al., 2012

12 3. Characterisation and role of the microbial ecosystem, especially protozoa In normal- or low-n diets, defaunation (or faunation with an Holotrich species) did not change rumen ammonia in sheep suggesting a better use of N by rumen microbes, which results in a lower urinary N The role of different protozoa in bacterial breakdown has been specified. Entodinium and Epidinium are especially active, whereas Holotrichs have a minor predatory activity. Therefore, lowering numbers of Entodinum and Epidinium species in the rumen may be a strategy for improving microbial synthesis (see next Figure) Cows are able to adapt themselves to fibrous diets by increasing the complexity of the rumen microbial community and the concentrations of protozoa, anaerobic fungi, and methanogens. On the contrary, rumen protozoa, fungi, methanogens and certain bacterial species are sensitive to N shortage which can explain the observed decrease in OM digestibility (Figure) The identification of key bacteria involved in protein metabolism by using DNA-Stable Isotope Probing was faced with strong methodological issues and results were inconsistent

13 The relative abundance of the 6 major protozoal groups in rumen of cattle and bacterial breakdown attributed to each of these protozoa groups. (Belanche et al., J. Anim. Sci) Effect of the level of protein and type of carbohydrate on the rumen concentration of certain microbial groups and their biodiversity. (Belanche et al., J. Nutr.) High Protein Low Protein FIB STA FIB STA Concentration (per g DM) Bacteria (mg) Protozoa (mg) Anaerobic fungi (µg) 1.70 a 1.40 b 1.58 ab 0.66 c Archaea (10 7 copies) Bacterial diversity 147 bc 148 b 152 a 138 c Fungal diversity 34 a 32 a 33 a 27 b

14 3. Characterisation and role of the microbial ecosystem, especially protozoa In normal- or low-n diets, defaunation (or faunation with an Holotrich species) did not change rumen ammonia in sheep suggesting a better use of N by rumen microbes, which results in a lower urinary N The role of different protozoa in bacterial breakdown has been specified. Entodinium and Epidinium are especially active, whereas Holotrichs have a minor predatory activity. Therefore, lowering numbers of Entodiniomorphids in the rumen may be a strategy for improving microbial synthesis (Figure) Cows are able to adapt themselves to fibrous diets by increasing the complexity of the rumen microbial community and the concentrations of protozoa, anaerobic fungi, and methanogens. On the contrary, rumen protozoa, fungi, methanogens and certain bacterial species are sensitive to N shortage which can explain the observed decrease in OM digestibility (Figure) The identification of key bacteria involved in protein metabolism by using DNA-Stable Isotope Probing was faced with large challenges and results were inconsistent

15 Take-home messages from WP2 Improved knowledge of the relation between rumen microbes (especially protozoa) and N ruminal metabolism Lowering dietary N below present recommendations decreases OM and fibre digestibility There is no adaptation to N underfeeding and microbial protein synthesis is not more efficient No real innovative way to decrease ruminal protein degradation was detected

16 Improving the nitrogen economy of the dairy cow Reducing Nitrogen Excretion WP3 Factors affecting the conversion of absorbed AA into milk protein Understanding the determinants of the efficiency of dietary Nitrogen Utilisation Chris K. Reynolds, Sophie Lemosquet, Isabelle Ortiques et al.

17 Milk N/Apparently digested N Feeding trial INRA Theix 0.65 EFFICIENCY OF N CONVERSION STARCH FIBRE Apparently digested N, g/d 11% improvement in N milk / N intake with high starch diets JDS submitted => Metabolism trial

18 MILK PROTEIN, g/d Main results so far 900 EFFICIENCY OF MP UTILIZATION improved with starch N = 48; 3 Rednex experiments WP WP3.4 Cantalapiedra-Hijar et al., 2012 (3R) EFFECTS OF STARCH Indications of higher microbial protein synthesis (purines in urine) JDS submitted Y = 225.4*** X Increased whole-body metabolic use of Leu (IRL) in favor of protein synthesis EAAP Y = 173.1*** X AVAILABLE MP, g PDIE/d 1600 ENERGY FIBER STARCH 1800

19 Objectives Univ. Reading Determine the effects of metabolisable protein (MP) supply fed both above and below metabolic requirement on post ruminal nutrient absorption and metabolism Determine the effects of different forage types (maize vs. grass silage) on post ruminal nutrient absorption and metabolism 19

20 Nitrogen Intake Barratt et al., Protein (Linear), P < 0.001; Forage, P <

21 Efficiency of N Dietary Use Barratt et al.,

22 Arterial urea concentration Barratt et al., Forage, P < 0.06; Protein (Linear), P <

23 Efficiency of N Dietary Use Milk Barratt et al.,

24 Conclusions Univ. Reading Effects of increased protein intake Linear increase in DMI, milk yield, milk N, rumen ammonia and PDV ammonia flux and arterial urea concentration Decrease in N efficiency with increasing dietary protein Effects of forage Higher N intake on grass silage based diets No forage effect on DMI or milk yield Increased rumen ammonia, PDV ammonia flux and arterial urea on grass silage based diets No forage effect on N efficiency (milk N/N intake) No forage protein interactions seen Clear positive relationship between N intake and rumen ammonia, arterial urea, and milk urea concentration, all of which are negatively related to N utilization efficiency Barratt et al.,

25 Research Questions - INRA Rennes 1. Does balancing the EAA profile increase milk protein yield and metabolisable protein (MP) efficiency both at low and high MP supply? 5 experiments: 3 balancing the whole EAA profile through duodenum infusions (4 to 6 cows) Are mammary uptakes of AA modified? 2 experiments with Lys, Met and Leu balanced through diet (16 to 32 cows): 2. Among the 9 EAA to balance are Val, Ile and Arg important?

26 Milk protein and efficiency increased when balancing AA profile at low and high MP supply Milk protein yield, g/d AA+ AA- Exp 1 Exp 2 Exp 3 Exp PDI, g/kg DM (CP from 13.5% to 19% of DM)

27 Mammary net uptake of only the limiting EAA increased: a higher waste of nitrogen when increasing both intestinal supplies of EAA and NEAA through increased MP LP HP + 72 g/d of N Rumen LP HP Intestine EAA: + 28 g/d of N NEAA: + 24 g/d of N AA Intestin AA AA- AA+ Intestine EAA: +28 g/d of N NEAA: - 28 g/d of N AA - AA+ = 0 g/d of N Mammary Uptake = Output EAA only: + 10 g/d of N AA AA Mammary Uptake = Output EAA only: + 12 g/d of N N efficiency N efficiency

28 Take home messages WP3 1. Dietary starch increases efficiency of dietary N utilization through effects on digestion and metabolism 2. Forage type has less of an effect when total rations are balanced for major components (starch, NDF, etc.) 3. Clear relationships between N intake and NPN metabolism and thus milk urea N is negatively correlated to N efficiency (this is relative to WP6). 4. Improving the balance of EAA provided to the mammary gland increases milk protein production across a range of metabolizable protein supplies. 5. Further work is needed on BCAA requirements 6. Masses of new knowledge of the metabolism of amino acids and other nutrients in lactating dairy cows fed diets below and above requirements for MP.

29 WP4 New feeding strategies improving N recycling while reducing N inputs Betina Amdisen Røjen, Niels Bastian Kristensen & Mogens Vestergaard Dept. of Animal Science, AU-Foulum, Aarhus University, Denmark Innovative and practical management approaches to reduce nitrogen excretion by ruminants

30 Increased N utilization by: 1. Reduced dietary N concentration 2. Increased blood to gut transport of urea-n N-efficient dairy cow

31 1. Optimize urea recycling to the GI tract while reducing N inputs 2. Ability of blood urea to sustain rumen ammonia from recycling with infrequent N supply 5. Competition between kidney and gut for urea with increased salt and water intake Kidney Blood urea-n Salivary urea-n Feed protein protein degradation Hindgut 3. Role of urea transporter proteins in urea transport across ruminal epithelia 4. Increased hindgut fermentation on urea recycling and N utilization

32 The total amount of urea transferred from blood to gut does not increase with decreasing N level The permeability of the gut epithelia for urea is up-regulated with reductions in N intake thus adapting to dietary conditions, but it is not up-regulated enough when N status of the cow get insufficient to sustain optimal microbial protein synthesis Net portal urea-n flux, mmol/h r = 0.185; P = Arterial urea-n concentration, mmol/l Ruminal extraction of arterial urea-n, % r = ; P < Arterial urea-n concentration, mmol/l

33 1. A Low-N diet leads to increased ruminal tissue permeability to urea 2. But immediate return of N in ammonia apparently not equilibrated with the rumen ammonia pool Ruminal extraction of arterial urea-n, % Low nitrogen diet High nitrogen diet Ruminal vein - arterial ammonia, mmol/l High N Low N Arterial urea-n concentration, mmol/l Time relative to urea infusion, min

34 Use of infrequent N supply (i.e., 6-h urea infusion, ) leads to sustained increase in arterial urea concentrations 9-16 h after end of infusion But there was no increase in urea recycling to the gut So, the cow was unable to make use of blood urea via urea recycling to sustain rumen ammonia concentrations during periods of the day where rumen N supply was at a minimum Arterial blood urea-n, mmol/l Time relative to feeding, h Ruminal ammonia concentration, mmol/l Time relative to feeding, h 6-h inf urea 24-h inf urea Water inf

35 Increased carbohydrate supply to the hindgut induced the predicted reduction in blood urea, but mainly through changes in ammonia fluxes - not by increased urea recycling to the hindgut Net portal flux, mmol/h ammonia Control Oligo urea

36 Take home message - WP4: Urea recycling is less efficient than hypothesized and difficult to manipulate to increase N efficiency in dairy cows This points to increased precision in dairy cattle nutrition as the most feasible short-term strategy to improve N efficiency

37 TMR / PMR Accuracy and precision obtained through biological monitoring system More use of information we already have available Silage Better accuracy and precision Need for new tools to monitor physiological status, nutritional sufficiency, and nutrient utilization COW 1 Feces Urine Milk Silage Premix Additive COW 2 COW 3 COW 4 Production Feces Urine Milk Feces Urine Milk Feces Urine Milk

38 Ways to reduce N excretion Reduce N intake Conclusions Adjust EAA composition in rations at a lower N intake Use starch-rich vs. fibre-rich rations Use of certain essential oils in the forage Reduce protozoa, especially large species Feed by-pass CHO to increase hindgut fermentation Use precision feeding of protein. and relevant management tools (see also later talks)

39 Thank you for your attention Questions? Culled organic dairy cows finishing their service as suckler cows for two bull calves on semi-natural pastures

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