AMINO ACID NUTRITION IN DAIRY ANIMALS AND ITS EFFECT ON MILK PRODUCTION AND MILK SOLIDS Dr Hinner Kster

Managing Director: Scinetic The fundamental goal of ruminant protein nutrition in dairy cows is to optimize the efficiency of utilization of dietary nitrogen (N) to maximize milk production per unit of N consumed. This requires fine-tuning of protein nutrition in two ways. Firstly, it requires feeding the correct types and amounts of ruminally degradable protein (RDP) that will meet but not exceed the requirements of ruminal microorganisms for N substrates as dictated by the types and amounts of ruminally fermentable carbohydrates in the diet. Secondly, it requires feeding ruminally undegraded dietary protein (UDP), with an amino acid balance that complements that of ruminally synthesized microbial protein in amounts which, when combined with microbial protein, provides the desired protein to energy ratio in absorbed nutrients. Like all mammals, ruminants have specific metabolic requirements for amino acids for maintenance and productive functions. Amino acids are uniquely essential as precursors for protein synthesis. They also serve for the synthesis of other N-containing metabolites as precursors for gluconeogenesis, and as sources of metabolic energy when oxidized to CO2. Improvement of ruminant protein nutrition is a matter of practical concern. The increasing attention that must be given to ration formulation to support increasing levels of milk production, the increasing concerns of waste N disposal, and the growing emphasis on milk protein production all provide incentives to seek ways to maximize efficiency of N utilization. Amino acids for ruminants Amino acids are an already proven concept in monogastric nutrition. Optimising the supplies of individual amino acids has been common practice in poultry nutrition for over 30 years and in pig nutrition for the last 20 years. The principal advantage of doing this is an increase in animal performance lean growth in broilers and growing pigs and egg production in laying hens, with an accompanying improvement in feed conversion efficiency. A similar improvement in performance may also be obtainable for ruminants by paying more attention to individual amino acid supplies, rather than trying to satisfy total amino acid supplies on a global basis. However, due to the role played by the rumen in transforming a large proportion of dietary protein into microbial protein, it has been very difficult to determine whether there is a significant performance advantage to increasing the supplies of specific individual amino acids postruminally for the dairy cow of high genetic merit. The amino acid balance concept for ruminants

First and foremost, a ruminant needs a sufficient supply of all amino acids, essential and non essential, at the systemic level, to satisfy the large majority of amino acid demands of maintenance and production. This is normally achieved by formulating rations to satisfy the recommendations laid down in any modern protein system such as the MP system (AFRC, 1992) or the protein digested in the intestine (PDI) system. Following the principles of monogastric nutrition, a further gain in performance can be envisaged if the correct balance or profile of digestible amino acids is supplied. The improvement in performance in monogastrics is achieved by increasing the efficiency of total amino acid utilization and reducing the energetic load for removing surplus amino acid N (Henry, 1993; Leclercq, 1996). Sources of absorbable amino acids Absorbable amino acids are provided by microbial protein synthesized in the rumen, UDP, and endogenous secretions. Ruminally synthesized microbial protein typically supplies 50% or more of the amino acids when production-type diets are fed. Microbial protein is supplied by predominantly fermentative populations of bacteria, protozoa, and fungi. Over 200 species of bacteria, more than 20 species of protozoa, and at least 12 species of fungi have been found in ruminal digesta. Bacteria generally constitute most of the microbial biomass in the rumen. Maximum yield of microbial protein can be obtained only if optimum quantities of energy, N, minerals, and other growth factors are supplied to the microbes. Availability of energy is thought to be the main regulating factor for the growth of anaerobic bacteria and, hence, for maximum microbial protein yield in the rumen. Imperfect knowledge of the amino acid requirements of ruminants and the paucity of information on the yield of microbial protein digested in the small intestine, make it impossible to estimate reliably the extent to which microbial protein meets amino acid requirements for lactation of dairy cows. It is well known that in different circumstances ammonia, amino acids, peptides, carbohydrates, branched chain acids or minerals may be limiting for microbial growth and that different microbial species have different efficiencies in the conversion of fermentable substrate into microbial protein (Bryant, 1973). Microbial protein is considered to be of high quality because of apparent high digestibility and an essential amino acid composition that is similar to that of lean body tissue and milk (Table 1). Endogenous Protein Endogenous secretions and abraded epithelial cells provide some absorbable amino acids but quantitative information describing this supply in ruminants is

Table 1. A comparison of the essential amino acid (EAA) profiles of lean tissue and milk with that of ruminal bacteria and protozoa and common feeds (% of total EAA) Source
Animal products 1 Lean tissue 2 Milk Rumen microbes 3 Bacteria 4 Bacteria 5 Protozoa 6 Forages Lucerne Maize silage Haycrop silage 6 Grains Barley Maize, yellow Maize gluten feed Oats Sorghum Wheat 6 Plant proteins Brewers grain Maize gluten meal Maize DDG w/ solubles Cottonseed meal Linseed meal Peanut meal Rapeseed meal Soybean meal Sunflower meal 6 Animal proteins Blood meal Feather meal Fish meal (menhaden) Meat meal Meat and bone meal Whey, dry
1
2

Arg 16.8 7.2 10.2 10.6 9.3 10.9 6.4 8.9 12.8 10.8 12.0 15.6 9.4 15.2 8.9 6.9 7.7 25.4 25.7 13.5 14.0 16.3 19.4 7.6 14.7 13.1 18.8 20.5 5.6

His 6.3 5.5 4.0 4.3 3.6 5.2 5.5 5.3 5.9 7.0 7.9 5.4 5.8 6.6 6.4 4.7 7.2 6.0 5.2 5.4 6.7 5.7 5.9 11.2 1.1 5.7 5.2 5.5 3.7

Ile 7.1 11.4 11.5 11.6 12.7 10.9 10.3 11.0 9.6 8.2 8.5 9.5 9.4 9.7 10.6 9.3 9.8 7.7 13.3 9.9 9.3 10.8 10.1 2.1 10.0 9.3 8.0 7.8 12.4

Leu 17.0 19.5 16.3 15.5 15.8 18.4 27.8 18.9 18.4 29.1 24.6 18.1 30.9 18.9 17.6 36.4 26.3 13.9 14.8 15.2 16.9 17.0 15.5 22.8 29.3 16.5 17.1 16.2 20.1

Lys 16.3 16.0 15.8 17.3 20.6 11.1 7.5 10.3 9.6 7.0 8.2 10.0 5.6 8.0 11.4 3.8 6.2 9.6 8.1 10.0 13.1 13.7 8.6 15.7 3.9 17.0 14.1 14.2 17.5

Met 5.1 5.5 5.2 4.9 4.2 3.8 4.8 3.8 4.5 5.0 4.6 4.3 4.3 4.6 4.8 5.5 5.2 3.8 3.5 2.4 4.8 3.1 5.4 2.1 2.1 6.3 3.7 3.6 4.3

Phe 8.9 10.0 10.2 10.0 10.7 12.2 12.0 13.5 13.3 11.3 10.1 11.5 12.6 12.6 10.3 13.8 11.1 12.2 11.1 11.5 9.5 11.0 11.0 12.3 10.0 8.8 9.7 9.2 7.4

Thr 9.9 8.9 11.7 11.0 10.5 10.6 10.1 10.3 9.1 8.4 9.6 9.2 8.0 8.3 11.4 7.5 10.3 7.7 8.9 6.5 10.5 8.6 9.1 8.1 10.5 9.5 9.1 9.0 13.2

Trp 2.5 3.0 2.7 2.6 2.8 3.4 1.4 3.3 3.1 1.7 1.6 3.2 2.2 3.4 3.0 1.5 2.7 2.9 3.5 2.8 3.0 3.0 2.8 2.7 1.5 2.4 2.0 1.8 3.8

Val 10.1 13.0 12.5 12.2 9.7 13.5 14.1 14.7 13.6 11.5 12.8 13.3 11.8 12.6 15.6 10.7 13.4 10.8 11.8 10.6 12.4 10.6 12.3 15.4 17.1 11.3 12.4 12.1 11.9

EAA , %CP

40.0 40.7

38.5 42.3 38.8 42.8 39.8 31.9 46.3 44.2 37.7 43.1 41.1 36.9 41.9 47.6 45.0 49.4 31.4 44.8 37.5 38.0 50.8

From Ainslie et al. (1993); average values of empty, whole body carcasses as reported in three studies. 2 Each value is an average of observations from Jacobson et al. (1970), McCance and Widdowson (1978), and Waghorn and Baldwin (1984). 3 From Clark et al. (1992); average values from 61 dietary treatments. 4 From Storm and Orskov (1983); average values from 62 literature reports. 5 From Storm and Orskov (1983); average values from 15 literature reports. 6 Calculated from values presented in European Amino Acid Table: first edition 1992 except for linseed meal, peanut meal, and feather meal that were calculated from values presented in Feedstuff Ingredient Analysis Table: 1991 edition.

extremely limited, except that it is influenced largely by intake and diet composition, for example the intake of digestible organic matter. UDP The second major source of absorbable amino acids is UDP. The ruminant does not have to rely solely on microbial protein since some dietary protein escapes rumen fermentation and passes to the lower gut undegraded. Therefore, the extent of dietary protein breakdown and the synthesis of microbial protein result in marked alterations in the quantity of pattern of amino acids absorbed from the gut of ruminants compared to the amino acid composition of the diet. This may influence greatly the yield and composition of milk produced in high producing dairy animals. The quantity of UDP flowing from the rumen for a feedstuff depends on the proportion of the total protein that is degradable, the rate at which the degradable fraction is degraded, and the residence time of the protein source in the rumen. Primary factors influencing one or more of these processes include microbial proteolytic activity in the rumen, microbial access to the protein, level of feed intake, the ingredient composition of the rest of the diet, physical form of the feedstuff, protein solubility, and ruminal pH. Proteolytic activity in the rumen surprisingly is quite variable and is influenced largely by dietary factors that affect the type and number of microorganisms in the rumen (Broderick et al., 1991). Nutritive value of UDP There are few direct estimates of the intestinal digestibility of UDP. The most direct approach is to increase the amount of UDP entering the duodenum in graded fashion while measuring the increment of UDP passing to the terminal ileum. A summary of several of these studies (Hvelplund, 1985; Rae and Smithard, 1985; de Boer et al., 1987; Frydrych, 1992) indicates that the UDPdigestibilities of most feed proteins are similar (80 to 90%). However, there are exceptions, such as soybean meal (SBM) and fish meal, where estimates of digestibility are generally higher than 90%, and rapeseed meal and meat and bone meal, where the estimates vary between 60 and 80%. Using a recently developed in vitro approach for both abomasal and intestinal digestion, Calsamiglia and Stern (1993) obtained digestibilities of the UDP in SBM, maize gluten meal, fish meal, blood meal, hydrolyzed feather meal, and meat and bone meal to be 89.8 2.6, 87.6 2.7, 85.4 2.6, 80.1 16.7, 69.5 3.9, and 54.0 6.2%, respectively. Clearly, differences in postruminal protein digestion between various sources must be considered when determining protein value for ruminants. Factors affecting intestinal balance of amino acids

Multivariate analysis of measurements of amino acid passage to the small intestine in ruminants (Rulquin and Vrit, 1993) confirmed that the amino acid composition of UDP and the proportional contribution that UDP makes to total protein flow accounts for most of the variation in amino acid balance of duodenal digesta. This would be expected because feed proteins vary greatly in amino acid composition and generally differ from microbial protein in amino acid composition (Table 1). As a consequence, the greatest influence of diet on amino acid profiles of duodenal digesta is seen when large amounts of high-UDP feeds with amino acid profiles most different from microbial protein (e.g., maize gluten meal, blood meal, and feather meal) are fed (Stern et al., 1983; Titgemeyer et al., 1989; Waltz et al., 1989). Lactational responses to postruminal administration of proteins and amino acids. As the quantity of milk produced per cow continues to increase, it will be increasingly more difficult to meet specific nutrient requirements of the lactating dairy cow. It is our belief that many cows producing in excess of 30 kg of milk daily fail to produce to their genetic capacity because of the lack of some key nutrient. This shortage of a specific nutrient may result because nutrient intake is limited by appetite or physical capacity of the animal or because of the inability of the rumen microbial system and/or tissues of the animal to generate sufficient quantities of precursors to meet the demands of the mammary gland for maximum synthesis of milk. If we are to improve the nutritional status of the high producing dairy cow, it is imperative that we identify those key nutrients which may be in short supply to the lactating gland and devise ways to supply these nutrients in times of greatest need. Results of postruminal infusion trials suggest that by-passing rumen fermentation with amino acids increases both milk protein yield and milk protein percent (Table 2). Ration protein source determines the quantity of milk protein secreted Rulquin and Vrit (1993), from a compilation of trials where treatment differences in CP content and N-degradability were negligible, showed that the protein source in the diet could have a marked influence on milk protein secretion. Soybean meal was superior to maize gluten meal and fishmeal was superior to soybean. In fact, fishmeal-containing diets outperformed maize gluten meal rations by as much as 150g of milk protein/cow/day. It would appear that the potential advantage of low protein degradability of maize gluten meal is compromised by its poor amino acid profile. The amino acid profile of the different protein sources would appear to have an important effect in determining the quantity of milk protein secreted.

Table 2. Daily milk yield and milk protein secreted by cows supplemented postruminally with casein with or without glucose (Adapted from Clark, 1975).
Diet Concentrate, alfalfagrass hay Concentrate, alfalfagrass hay Concentrate, alfalfagrass hay Concentrate, maize silage (urea included) legume-grass hay Concentrate, maize silage, hay Concentrate, maize silage, hay Concentrate, hay CP in diet, % 16.5 15.0 17.0 17.0 16.0 Infusate Na-caseinate Na-caseinate Na-caseinate Na-caseinate and glucose Na-caseinate + Met Casein Casein + Met Casein + Met + glucose Na-caseinate + Met Na-caseinate + Met Na-caseinate + Met Na-caseinate + Met Casein Casein Casein Casein Na-caseinate Na-caseinate Na-caseinate Change in milk yield, kg 1.3*** 1.6** 2.2* 2.8* 1.6 Change in milk protein yield, % +11*** +14** +11** +15** +12**

16.4 15.0 15.0 16.2

1.0** 1.7* 1.1* -.2 -1.0 -.1 .4 3.0 2.0 .4 1.8*** 4.2 3.9 2.9

+ 9** +13** +16** + 1 -3 +6 +12**

Concentrate, maize silage, (urea included), hay Concentrate, maize silage (urea included) Concentrate (urea include), straw
*P<.10. **P<.05. ***P<.01.

16.1

17.0 17.0 17.0

+8 +16*** +31 +16 +22

Casein has been the mostly widely used protein source to investigate the general interest in improving the quality of protein available to the dairy cow. Consistently large responses to casein infusions have been observed in terms of milk yield (1 to 4 kg per cow/day) and milk protein yield (50 to 180 g per cow/day) (Clark,1975). There is apparently both an important quantitative as well as qualitative component to the initiation of this response. A large part of the volume response may be attributable to an increase in the total quantity of amino acids absorbed (PDI) by the dairy cow. It is also evident that the responses are very much related to the basal level of protein supply. When protein supplies are lower than requirements, a larger proportion of the additional available amino

acid-N is partitioned towards milk protein synthesis relative to other productive functions compared with when requirements are already satisfied (Whitelaw, et al., 1986). However, the qualitative advantage of casein, due to its balanced amino acid profile, is also important, as demonstrated in trials comparing isonitrogenous quantities of casein and soya isolate (Choung and Chamberlain, 1993). Whereas 50% of the extra amino acid-N from casein was found in milk protein, only 20% was recovered as milk protein when soya isolate was infused. Methionine and lysine, the first two limiting amino acids Many trials have tried to identify the sequence of limiting amino acids for milk protein production by infusing individual amino acids post-ruminally. Schwab, et al. (1976) demonstrated that methionine and lysine were likely to be the two most limiting amino acids in dairy rations for milk protein secretion. Increasing supplies of these two amino acids alone improved milk protein content by 1 g/kg and milk protein secretion by 45 g/day. Many experiments of a similar nature have since been carried out over a wide range of diets with the focus of the investigations being the response to increasing digestible lysine (lysDI) and methionine (metDI) supplies together. Rulquin (1992) summarized much of this work (Table 3) and showed quite clearly that in the vast majority of rations, lysine and methionine must be the first two limiting amino acids. By increasing these two amino acids alone, milk protein was increased on average by 35 and 50 g/head/day in mid and early lactation cows respectively. Table 3. Effects of the stage of lactation on the responses to a post-ruminal supply of methionine and lysine Stage of lactation Early lactation Mid lactation SED Week No. of trials Amino acids supplies (g/day) Methionine Lysine Productions Milk (kg/d) Protein (g/d) Fat (g/d) Milk composition Protein level (g/d) Fat level (g/kg) 1 to 9 (16) 10 to 29 (71)

10 24

8 20

+0.7 +56 +10

+0.1 +31 +1

0.9 51

+1.2 +0.5

+1.0 -0.0

0.7 1.6

What is further curious is the apparent influence of balancing rations for lysDI and metDI on overall feed utilization. For 12 early-lactation trials analysed there was an improvement in apparent feed efficiency ((feed DM input/milk DM output (milk fat + milk protein)) in every case (Table 4). This could not be explained solely by an increase in milk protein output. In the trials where milk volume was improved, milk fat content was not reduced, thus milk fat secretion was also increased. In fact milk component yield (fat + protein) was often increased by at least 150 g/cow/day with an increase in apparent feed efficiency of 0.04 to 0.08. Practical approach to formulating in digestible amino acids for dairy cows All raw materials can contribute to the lysDI and metDI needs of the dairy cow. It is the judicious choice of raw materials and the proportions incorporated into concentrate formulations which will be important in achieving the required overall levels of digestible lysine and methionine. In order to formulate as accurately as possible it is important to have a reliable database of lysDI and metDI values for all the commonly used raw materials. For raw materials not present in tables, and for those recognized to be inherently variable, new lysDI and metDI values can be derived from measuring rumen N-degradability in sacco and from measuring the amino acid profile in the original raw materials. Table 4. Influence of ration lysDI/metDI concentration on milk performance and feed efficiency (Adapted from Sloan, 1997)
Original Reference Trial duration (days of lactation Ration lysDI/metDI concentration (% PDIE) Increase in intake (kg DM/cow/day) Increase in milk fat plus protein yield (g/cow/day) (Supplemented-Control) -0.1 131 Apparent feed efficiency kg DM intake/kg milk DM (fat plus protein output)

Control Robert et al, unpublished Brunschwig et al 1994 Brunschwig et al 1995 Socha et al 1994d Rulquin unpublished Chilliard et al unpublished 0-84 8.0/1.7

Supplemented 6.9/2.4

Control 7.35

Supplemented 6.92

21-140

6.9/1.8

6.9/2.4

0.4

149

8.67

8.29

0-56

6.9/1.7

6.9/2.1

-1.1

47

7.27

6.72

0-105 0-105 0-28

6.5/1.8 6.5/1.8 6.9/1.7

6.5/2.2 7.0/2.2 7.7/2.1

-1.0 0.8 0.4

-32 158 146

9.06 9.06 7.48

8.80 8.85 7.21

5-42 5.26 5-26 27-305 27-305

6.9/1.6 6.9/1.6 6.9/1.6 6.2/1.9 6.2/2.0

6.9/2.2 6.8/2.8 8.2/2.6 6.8/2.2 6.8/2.2

0.1 1.1 1.7 1.0 0.0

291 269 133 150 120

6.48 6.20 6.20 10.73 10.84

5.87 5.99 6.56 10.40 10.27

Robinson et al 1995

Actual milk production values no adjustment by covariance

The biggest breakthrough in practical amino acid nutrition of dairy cows was the compilation of all the relevant research work and the development of a systematic approach to estimating raw material values for digestible methionine and lysine contents and the publication of estimates for digestible lysine and methionine requirements. From this, lysine and methionine requirements for the lactating dairy cow were estimated to be 7.3 % and 2.5 % of total digestible amino acid supply (PDIE) respectively. Socha and Schwab (1994), using a similar approach but based on the CNCPS (OConnor et al., 1993), estimated requirements for lysine as 16.8% of total essential amino acids. Specific dose response studies (Robert et al., 1996) have confirmed the estimation of the lysDI requirements derived by Rulquin and Vrit (1993). A precise estimate of metDI requirements has been much more difficult to establish. The marginal response to additional metDI remained positive and linear between 1.5 to 2.35 (% of PDIE) metDI where the ration provided recommended levels of lysDI (7.3 % of PDIE) (Pisulewski et al., 1996; Socha et al., 1994a,b). However, Socha et al. (1994c) found no response to increased metDI in mid to late lactation cows. Where the lysDI level was only 6.50 % of PDIE no significant increase in milk protein secretion has been noted from additional metDI, whereas increasing simultaneously lysDI and metDI from 6.50/1.80 to a minimum of 6.80/2.15, increased milk protein secretion by 37 g/cow/day (Sloan et al., 1994) The estimates of 2.5 % and 7.3 % of PDIE proposed by Rulquin and Vrit (1993) would, however, appear to be very good first approximations of metDI and lysDI requirements respectively. In countries where this approach to rationing cows in terms of lysDI and metDI as a % of PDIE has been adopted, a reasonable compromise between optimising milk protein secretion and the cost of doing so, has resulted in the adoption of practical recommendations of 7.0 lysDI and 2.2 metDI (as a % of PDIE). In the approach proposed by Rulquin and Vrit (1993), each raw material is assigned a lysDI and metDI value which takes into account both the microbial contribution calculated form the quantity of fermentable organic matter (FOM) digested in the rumen and the bypass contribution. All forages have relatively high lysDI values (6.8 to 7.0 % of PDIE) and above average metDI values (1.9 to 2.0 % of PDIE). This is due to the majority of the contribution to lysDI and metDI being of microbial origin. In general, rumen degradabilities of forages are high, thus the digestible amino acid contribution from bypass protein is minor. Likewise for cereals, lysDI (% of PDIE) levels tend to be slightly above average, due to their low crude protein content and high fermentable organic matter content, which again favours the microbial digestible amino acid contribution. Maize grain is an exception. Although its CP content is low, rumen N-degradability is very low (~ 0.40) and at least 0.2 of the FOM escapes rumen fermentation. These factors maximise the bypass digestible amino acid contribution for this raw material. With its low lysine content this results in a low value of lysDI (% of PDIE) for maize and, in general, for maize-based byproducts. Maize gluten meal has the

distinction of being the raw material with the lowest lysDI concentration because of its high CP content and extremely low degradability (0.28). On the other hand, maize gluten feed has a lysDI value of 6.4 (% of PDIE) which is average. This is due to the high N-degradability (~ 0.69) of this raw material. The raw materials that can most influence the profile of absorbed protein (PDIE) are concentrated sources of bypass protein. All protein-rich raw materials, particularly those with a low N-degradability, have the potential to modify considerably the amino acid profile of absorbed protein. Soybean meal is above average in lysDI (7.0 % of PDIE). However all the other proteinaceous meals of vegetable origin tend to be low in both lysDI and metDI. Only animal protein sources have levels of either of the potential limiting amino acids superior to those required in the overall ration. Blood meal (banned in the European Union) is an excellent source of lysDI, but fish meal is the only raw material that is an excellent source of both lysDI and metDI as a % of PDIE. In practical ration formulation it is impossible to meet both the lysDI and metDI constraints of 7.0 and 2.2 (as a % of PDIE) respectively from conventional raw materials alone; cows cannot be fed entirely on fishmeal. Unfortunately, feeding synthetic amino acids is not a potential solution. Neither DL-methionine nor the hydroxy analogue of methionine, the two commercially available forms of synthetic methionine, resist rumen degradation (Loerch and Oke, 1989) and thus are not sources of bypass methionine to increase ration metDI levels. The recent availability of good quality sources of rumen protected amino acids however not only permits the lysDI and metDI constraints to be met but also gives the formulator greater choice and flexibility in the other raw materials that can be incorporated to attain these levels. Obviously balancing dairy rations uniquely for lysine and methionine is not sufficient to achieve maximum milk performance. The comparisons of infusions of essential amino acids versus casein showed that the first two limiting amino acids could account for 40 to 50% of the total response to casein. Thereafter other amino acids become limiting. Various amino acids have been suggested as being potentially 3rd or 4th limiting. Histidine appears to be a good candidate. Its theoretically low concentration in microbial protein would apparently suggest that mammary supplies of histidine for milk protein synthesis often look limited. In the early work of Schwab et al. (1976) although lysine and methionine were clearly first limiting, infusion of extra histidine, in addition, tended to increase milk protein secretion through a milk volume response. More recently some Finnish work (Huhtanen et al., 1996) indicated the potential importance of histidine as a first limiting amino acid on grass silage, cereal/sugarbeet pulp based diet where urea was the only supplementary source of crude protein. Another amino acid often thought to be potentially limiting on grass silage/hay based diets is leucine. Isoleucine has also been suggested as a potential limiting

amino acid. The non-essential amino acids are normally discounted as being potentially limiting for milk protein synthesis. In practice, as we do not yet formulate to fully meet the recommendations for the two first limiting amino acids lysine and methionine, there is little point in trying to incorporate formulation constraints for the 3rd and 4th limiting amino acids on the limited knowledge we have available today. However it is important for the future that the sequence of limitations is more clearly identified. In pigs the identification of threonine and tryptophan as 3rd and 4th limiting amino acids has given the pig nutritionist the opportunity to reduce CP inputs whilst maintaining performance. Responses to amino acids largely independent of ration energy level and source In pigs, lysine requirements are expressed with respect to net energy (Fuller, 1994); energy being needed for the process of tissue protein synthesis. Erfle and Fisher (1977) commented that an adequate energy intake also had to be provided in dairy cows to permit a response to intravenous infusions of lysine and methionine. However, where energy intakes have been varied between 90 and 110 % of conventional recommendations there has been no noticeable influence on the magnitude of the response to additional digestible methionine and lysine (Rulquin and Delaby, 1994; Brunschwig et al., 1995). In fact, Colin-Schoellen et al. (1995) observed that the improvement in milk protein secretion to extra metDI and lysDI tended to be higher (88 vs 41 g/cow/day) at low versus high energy intakes (90 vs 112% of recommendations). In trials where the extra energy was supplied in the form of fat (Christensen et al., 1994; Canale et al., 1990) the effects on milk protein content of supplying extra metDI and lysDI were significant and comparable to those obtained on a diet with no added fat. The general negative effect of added fat, independent of source, on milk protein content was counterbalanced by the positive effect of providing extra digestible lysine and methionine. The need for rumen protected proteins There are several reasons and advantages for decreasing the rate and extent of ruminal degradation of selected feed proteins and for having a variety of highUDP feeds available to the producer. First, there are feeding situations in which the diet does not contain adequate quantities of absorbable amino acids relative to the supply of absorbable energy. This can occur because of: (1) too many ingredients in the diet which contain insufficient UDP relative to RDP, (2) some limitation on ruminal synthesis of microbial protein (e.g. insufficient supplies of fermentable carbohydrates or of RDP), (3) feeding supplemental fats which are dense sources of metabolizable energy but which are not used for microbial cell growth, or (4) some dependence on mobilized body fat for metabolizable energy, such as in the early lactation dairy cow. Second, high quality pastures, hays, and

particularly silages often contain so much RDP that any required protein supplementation must be limited to high UDP feeds if the diet is to be balanced for both RDP and UDP. And third, the use of higher UDP feeds permits greater use of NPN supplements and less reliance on more degradable sources of true protein for ruminal synthesis of microbial protein. It also must be remembered that requirements for absorbable amino acids, relative to absorbable energy, are highest when ruminants are fed for maximum milk protein production and that metabolism enhancers such as bovine somatotropin may further increase amino acids requirements. One approach to increasing the contribution of UDP to total CP in the diet, as alluded to already, is to feed high-protein byproduct feeds (e.g. maize gluten, meat, hydrolyzed feather, fish, and blood meals) that are already of lower ruminal degradability. However, cost, commercial availability, uniformity, amino acid balance and intestinal digestibility and palatability of product dictate their use. The alternate approach is to reduce artificially the rate of ruminal degradation of high protein forages, oilseeds, and oilseed meals that possess good amino acid balance and good intestinal availability but which are rapidly degraded. This approach: (1) increases the number of higher UDP feeds with which to optimize diets for UDP/RDP ratios, intestinal amino acid balance, and price, (2) has the advantage of being applied to feeds of choice and of providing controlled ruminal protection so as to maximize protection with minimal reduction in intestinal digestion, and (3) reduces the reliance on byproduct feeds as the only high UDP feed sources. The need for rumen protected amino acids There is a need for ruminally protected forms of Lys (RPLys) and Met (RPMet) because most feeds contain low amounts of Lys, Met, or both. In turn, the need is greatest when rations are balanced for high levels of production and UDP provides a larger portion of total absorbable amino acids. This is most evident by comparing the required contributions of Lys and Met to total essential amino acids in duodenal digesta of lactating dairy cows (Table 5) with measurements of the same when cows were fed a variety of maize-based diets (Table 6). Collectively, the data presented in Tables 1, 5 and 6 indicate the difficulty of meeting the desired levels of both Lys and Met in duodenal digesta when maizebased diets are fed. Moreover, the data indicate that complementary sources of supplemental UDP can be selected to achieve the desired Lys to Met ratio in duodenal digesta of lactating cows but levels of neither will be high enough in total ration UDP to optimize intestinal amino acid balance. It is expected from studies designed to determine the sequence of amino acid limitation in ruminants (Schwab et al., 1976; Fraser et al., 1991) along with the compositional data presented in Table 10 that balancing UDP for other essential amino acids can be accomplished by complementary feed proteins without the use of large amounts

of high-UDP, unbalanced feed proteins (e.g. maize gluten meal, blood meal, and feather meal). Table 5. Determination of the required contributions (%) of Lys and Met to total EAA1 in duodenal digesta of lactating dairy cows for maximum yield of milk protein when cows consumed conventional maize-based diets2 (Adapted from Schwab, 1995). Lysine 14.9 14.8 15.2 14.0 14.5 14.7
1

Original Reference Rulquin et al. (1990)3 Schwab et al. (1992)4

Met >5.1

Original Reference Rulquin et al. (unpublished)5 Socha and Schwab6 (unpublished)

>5.3 5.3

Average

>5.3

Average

Essential amino acids; includes Arg, His, Ile, Leu, Lys, Met, Phe, Thr, and Val. 2 The experiments involved graded infusions of Lys (in the presence of constant supplemental Met) and Met (in the presence of constant supplemental Lys) into the duodenum with simultaneous measurement of milk and milk protein production and AA flows to the small intestine. 3 -1 Midlactation cows were injected with 30 mg d bST and fed protein at 90 or 110 % of CP requirements. Each experiment was a replicated 4 x 4 Latin square. 4 Four cows were assigned to 4 x 4 Latin squares at 4 wk, 14-16 wk, and 21-23 wk post calving. Rations were balanced to meet 90-100% of CP requirements 5 Five cows were assigned to a 5 x 5 Latin square at 4-6 wk post calving. The ration was formulated to meet 115% of CP requirements. 6 Five cows were assigned to 5 x 5 Latin squares at 2 wk and 11-13 wk post calving. Rations were formulated to meet or slightly exceed protein requirements.

CONCLUSION Research clearly shows that paying more attention to the balance of amino acids absorbed by dairy cows has the potential to improve milk protein yield and certain other aspects of milk performance. 1. Milk protein content can be increased immediately in cows at any stage of lactation, by 1 to 2 g/kg. 2. Milk yield can be improved by up to 2.5 kg/day in early lactation (first 100 days). 3. Milk protein yield increase of 60 to 100 g/day over a complete lactation is attainable. 4. Feed efficiency can be improved by 0.05. To optimize these potential benefits it is essential to formulate dairy rations to contain higher levels of both lysDI and metDI than is currently practiced. The approach and recommendations proposed by Rulquin and Vrit (1993) are sufficiently robust that they can be used satisfactorily in a wide variety of dietary

situations. The challenges for the future will be to refine these recommendations still further with respect to stage of lactation and milk yield potential and to determine recommendations for the dietary levels of the other essential amino acids in order to improve the efficiency of utilization of dietary protein inputs. Table 6. Comparison of seven studies of the effect of ration composition (excluding fat, mineral and vitamin supplement) on measured passage of Lys and Met to the duodenum of Holstein cows during the first 150 d of lactation. 12 References2 3 4 5 10.0 25.0 25.0 40.9 26.2 14.2 24.7 9.4 28.7 25.5 30.0 20.0 40.0 13.3 28.5 27.1

Item Diet ingredients (% DM) Alfalfa hay Alfalfa silage Maize silage Grass-legume silage Maize Wheat byproducts Beet pulp Soyhulls Soybean meal Roasted soybeans Whole cottonseed Distillers grains Brewers grains Maize gluten meal Blood meal Feather meal Fish meal Meat meal Animal/fish blend Urea Diet CP (% DM) Flow to duodenum (g d-1) Total EAA Lys Met Flow to duodenum (% EAA) Lys Met
1 2 -1

6 26.0 5.0 30.2 12.3

7 45.4

34.6

9.0

12.5 16.1

4.0

9.8 .9 2.1 .3 1.5 .6 .2 16.2 17.3

4.7

4.0 8.5 4.8 9.0 1.8

8.3 4.0 6.0 9.0 1.8 8.0

4.0 1.5

17.3

18.1

19.6

19.7

19.0 1924 246 63 13.9 4.6

1619 1353 1384 1395 1840 1836 217 179 194 164 249 235 61 55 61 52 64 64 13.4 3.8 13.2 4.1 14.0 4.4 11.8 3.7 14.1 4.5 13.7 4.6

Intakes of DM averaged 21.3 kg d kg (range = 18.2 to 23.5). References: (1) Christensen et al., 1993; (2) Schwab et al., 1992; (3) Cunningham et al., 1993; (4) Klusmeyer et al., 1991; and (5, 6 and 7) Cunningham et al., 1991.

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