FIBER DIGESTIBILITY AND FORAGE FRAGILITY IN DAIRY CATTLE. K. Cotanch and R. Grant William H. Miner Agricultural Research Institute Chazy, NY

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FIBER DIGESTIBILITY AND FORAGE FRAGILITY IN DAIRY CATTLE K. Cotanch and R. Grant William H. Miner Agricultural Research Institute Chazy, NY Physically Effective Fiber System INTRODUCTION Mertens (1997) defined physically effective NDF (pendf) as the fraction of NDF that stimulated chewing and contributed to a ruminal digesta mat. He also proposed a standard laboratory method for measuring pendf that involved dry sieving and measuring the fraction of sample NDF retained on the 1.18-mm sieve. This large particle pool requires rumination and has a high resistance to escape from the rumen of sheep (Poppi et al., 1980). Mertens (1997) summarized a large data set that showed a positive correlation between pendf measured by dry sieving, ruminal ph, and milk fat percentage in dairy cattle. Based on Mertens (1997) data set, the minimal requirement for pendf in lactating dairy cows has been defined as approximately 21 to 23% of ration dry matter (DM) to maintain ruminal ph above 6.0 and milk fat above 3.4% for Holsteins. Since 1997, the pendf system has become widely used within the CPM-Dairy and Cornell Net Carbohydrate Protein System (CNCPS) ration formulation models to predict the effect of forage particle size on cow chewing response and ruminal ph. A problem has been lack of an on-farm tool that accurately measures pendf and results in physical effectiveness factors (pef) similar to those obtained by the standard dry sieving method. Recently, we developed the Z-Box to effectively measure pendf for common forages and total mixed rations (TMR) in on-farm situations (Cotanch and Grant, 2006). Particle Size versus Fragility A major assumption of the pendf system is that forage particle size explains all of the variation in chewing response. This assumption is not always true because we know that forages of similar particle length may elicit substantially different chewing times per kilogram of NDF. For example, oat straw may elicit nearly twice as much chewing as alfalfa hay at a similar particle size (data summarized in Mertens, 1997). Grass and alfalfa hays resulted in a range of 111 to 152 minutes of chewing/kg NDF compared to nearly 200 minutes of chewing/kg NDF from oat straw. This has been a recognized problem within the pendf system and currently nutritionists use their best judgment to adjust rations based on cow response to compensate for differences among forage sources in their ability to stimulate chewing. Several methods have been reported for measuring relative differences in forage particle fragility. Years ago, Troelson and Bigsby (1964) devised a mechanism to specifically examine forage particle size distribution after artificial mastication. More recently, two commonly discussed methods are: 1) comminution energy which is the

energy required to grind a sample through a mill, and 2) shear force which measures the force needed for a blade to pass through the forage stem using a Warner-Bratzler or similar machine common in meat science laboratories. Casler et al. (1996) has developed a particle size reduction index using a ball mill to grind leaf particles. The percentage of particles passing through a 1-mm screen upon milling is considered to be an index of forage fragility. Although it has not been investigated, Mertens (1997) and others have suggested that differences in forage fragility, or related characteristics such as stem brittleness, may explain the variation in chewing response not explained by particle size. Fragility is defined as the relative rate at which forage is reduced in particle size during chewing or some laboratory simulation of chewing action. Fragility may be related to lignin content and digestibility as well as to anatomical differences among plant species such as cell wall thickness (Van Soest, 1994). Consequently, digestibility of the forage cell wall may be predictive of forage fragility. As an example, Minson (1990) summarized data for grasses that related composition with resistance to comminution and voluntary intake in sheep. The grasses were categorized as high (81%), medium (72%), and low (56%) DM digestibility. As digestibility of the grass increased, NDF content decreased, grinding energy decreased, eating and ruminating times decreased, and voluntary intake increased. PRELIMINARY RESEARCH: NDF DIGESTIBILITY AND FORAGE FRAGILITY Miner Institute recently developed a ball milling method to assess the fragility of a wide range of forages (Cotanch et al., 2007). Briefly, forage samples are dried and placed in a ball mill loaded with ceramic balls because unpublished observations by Dr. Jim Welch at the University of Vermont had indicated that this process mimicked the grinding action of molar teeth. The forage samples are sieved using the standard method for measurement of pef prior to ball milling (pefi) and again following ball milling (pef BM ). Fragility is determined as the change in pef value (proportion of particles >1.18- mm sieve as determined by dry vertical sieving) of the ball-milled forage from the original sample: (pef i pef BM )/pef i x 100% The change in pef, or fragility, versus 24-h in vitro NDF digestibility for a range of forages is shown in Figure 1. The R 2 indicates that the 24-h NDF digestibility explains about one-half of the variation in forage fragility. The potential exists to combine a fragility factor, related to NDF digestibility, with the pef derived by sieving to arrive at a superior value to predict cow chewing response.

f % Change in pe 45 40 35 30 25 20 15 10 5 0 y = 0.4245x + 12.481 R 2 = 0.4616 0 20 40 60 80 NDFd24 Figure 1. Relationship of the 24-h in vitro NDF digestibility of various forages with the fragility of the forages as measured by change in physical effectiveness factor (pef) following ball milling. FORAGE FRAGILITY AND FIBER DIGESTIBILITY The final sample set included a variety of forages to encompass the range of forage types typical of northeast US and Japanese dairies. Grass silage ½ was a late 1 st, grass silage 3 was an early first, corn silage 7 was from crop year 2005 (high quality), while corn silage 4 was from crop year 2006 (low quality). The alfalfa silage was 3 rd from 2006, the oat hay was of high quality. The alfalfa hay stems and leaves were separated from the same hay sample. The Ballard grass hay 1 st and 2 nd were from the same field, harvested in 2006. Lastly, a wheat straw sample was included. Unfortunately there was not any BMR corn silage available for this part of the trial. There was not any pure timothy or reed canary grass hay available either, only the mixed native grasses. It should be noted that the hays and straw required special care for ball milling. These long forages tended to tangle and ball-up, and just roll around in the jar without being ground. This required that they be hand to approximately 5 cm to prevent tangling and allow independent movement and contact with the ceramic balls. However, with hand ting, particle separation became an issue especially with the leaf and stems of the alfalfa hay. To avoid particle separation, the hays were sieved through the Penn State particle separator and only particles retained on the top 2 screens (>8mm) were used for analyses. The alfalfa hay leaves and stems were analyzed as separate samples. All samples were analyzed in triplicate for NDF, ADF, ADL, and NDFd 24 with the Ankom Daisy system. Ash was determined by muffle furnace, 600 C for 3 hr. Table 1 lists the standard fiber analysis profiles of the forages.

Table 1. Standard analysis profiles including ADF, AD-lignin, NDF, ash and in vitro NDF digestibility at 24 hours. Forage %DM %NDF %ADF %Lignin Ash DMDt 24 NDFd 24 GS Bunk ½ 1 st 36.8 64.6 41.6 7.6 7.19 64.8 45.6 GS Bunk 3 1 st 32.1 61.8 38.5 5.8 6.96 72.7 55.8 CS Bunk 4 29.4 37.1 23.4 3.8 3.80 76.1 35.4 CS Bunk 7 27.4 44.3 26.2 3.4 2.39 74.7 42.8 LS Agbag 3 rd 40.7 50.2 32.6 5.7 10.03 82.0 64.2 Coarse Oat Hay 84.3 64.5 41.2 6.5 5.47 60.2 38.3 Alfalfa Hay Stems 90.0 56.3 45.9 10.8 6.97 59.5 28.1 Alfalfa Hay Leaves 89.2 21.4 16.9 4.5 11.76 88.8 47.6 Grass Hay Ballard 1 st 90.4 73.0 47.4 9.0 6.93 48.6 29.6 Grass Hay Ballard 2 nd 91.3 62.7 35.0 4.5 6.59 75.2 60.4 Wheat Straw 92.2 84.2 61.5 13.4 5.75 34.0 21.5 Each forage was sampled for original intact pef value as determined by dry sieving. Samples were ball milled in duplicate. Table 2 shows the reduction in pef value resulting from ball milling. The values are expressed as percentage of original pef and percentage reduction from the original sample. Table 2. The pef values of intact original sample and 6-hour ball milled (BM) sample with percentage decrease in pef value. Forage NDFd 24 Original 6-hour BM pef as % Change pef pef % of original pef GS Bunk ½ 1 st 45.6 61.1 36.9 60.4-39.6 GS Bunk 3 1 st 55.8 60.1 39.3 65.4-34.5 CS Bunk 4 35.4 88.9 67.5 75.9-24.1 CS Bunk 7 42.8 92.0 67.0 72.8-27.2 LS Agbag 3 rd 64.2 73.6 48.1 65.4-34.6 Coarse Oat Hay 38.3 86.3 50.8 58.9-41.2 Alfalfa Hay Stems 28.1 95.0 71.6 75.4-24.6 Alfalfa Hay Leaves 47.6 41.1 13.0 31.6-68.5 Grass Hay Ballard 1 st 29.6 84.1 63.1 75.0-25.0 Grass Hay Ballard 2 nd 60.4 91.5 76.7 83.8-16.1 Wheat Straw 21.5 99.7 84.8 85.1-14.9 It was theorized that the more fragile the forage fiber the greater the decrease in pef of the ball milled sample. Therefore a greater change in pef value resulting from ball milling, would indicate greater NDF digestibility. As seen in Figure 2 there is little relationship between fragility or pef reduction and NDFd 24, with y = 0.3112x + 18.569 and R 2 =0.08.

80 70 60 % Change in pef 50 40 30 20 10 0 y = 0.3112x + 18.569 R 2 = 0.0822 0 10 20 30 40 50 60 70 NDFd 24 Figure 2. Forage fragility as indicated by percentage change in pef plotted by NDFd 24. However, 2 outlier data points could be removed in order to improve the regression equation. The two samples are the alfalfa leaves and the 2 nd Ballard grass hay. The alfalfa leaves are a physiologically unique tissue compared to the other samples, and the Ballard grass hay 2 nd, showed tangling of the forage particles even after hand ting prior to ball milling. Removal of these two samples from the data set results in an improved prediction equation: y = 0.4245x + 12.481, R 2 = 0.46, as seen in Figure 1. In this sample set, the grass silage ½, grass silage 3, and oat hay all have similar NDF content of ~65%, but vary in their pef value after 6 hours of ball milling, from 37 to 51%. These results support previous research (Casler et al. 1996), indicating some relationship between forage fragility and forage digestibility. Though it remains to be seen how fragility or particle size reduction as determined with this ball mill method relates to animal chewing behavior. RATIONALE FOR CHEWING RESPONSE STUDY WITH CATTLE We proposed to measure chewing response to forages of similar particle size that represented the extremes shown in the data set of Figure 1. Four forages were evaluated: 1) a low NDF digestibility grass hay with a low fragility value, and 2) a high NDF digestibility grass hay with a high fragility value. These forages fit the expected positive relationship between NDF digestibility and forage fragility. The final two grass hays contained similar NDF digestibility, in the middle of the digestibility range (~40%), with 3) a high fragility value, and 4) a low fragility value. These forages did not fit the general relationship since they have similar NDF digestibilities yet substantially different fragility as measured by ball milling. We chose to use grass hays in this study because we did not want to confound our chewing results by: 1) different plant species (grass, legume, corn silage, cereal grains), 2) moisture content (dry hay, silages), or 3) genetics

such as brown midrib (bmr). Using test forages such as poor quality straw and bmr corn silage would have likely expanded the range in NDF digestibility and fragility, but we would have been far less able to control particle size and we would not have been able to assess the separate and potentially important effects of moisture, etc. We chose to feed the hays to heifers rather than adult dairy cows primarily because of the limited supply of these grass hays. Fortunately, at least one reference indicates that the relative rumination response for larger heifers such as we used will be similar to adult cows (Welch and Hooper, 1988). Results from this study have not been completely summarized and evaluated at this point. The overall purpose of the study will be to determine if the fragility factor assessed by ball milling would be of any practical use in adjusting pef values to better predict the cow s chewing response the ultimate bioassay for the pendf system. In addition, we will learn more about the interactions among forage NDF digestibility, forage fragility, and cow chewing response. Only with cow chewing response can we know for sure if a relationship between NDF digestibility and fragility will improve on the pendf system. REFERENCES Casler, M.D., D.K. Schneider, and D.K.Combs. 1996. Development and application of a selection criterion for particle size breakdown of smooth bromegrass leaves. Anim. Feed. Sci. Tech 61:57-71. Cotanch, K. W., and R. J. Grant. 2006. Measuring physically effective fiber on-farm to predict cow response. pp151-160 in Proc. 2006 Cornell Nutr. Conf. for Feed Manufacturers. October 24-26. Syracuse, NY. Cotanch, K. W., R. J. Grant, J. Darrah, H. M. Wolford, and T. Eguchi. 2007. Development of a method for measuring forage fragility. J. Dairy Sci. 90 (Suppl. 1): 563 (Abstr.). Goering, H. K., and P. J. Van Soest. 1970. Forage fiber analyses (apparatus, reagents, procedures, and some applications). Agric. Handbook No. 379. ARS-USDA, Washington DC. Grant, R. J., and K. W. Cotanch. 2005. Physically effective fiber for dairy cows: current perspectives. pp 61-70 in Proc. Cornell Nutr. Conf. for Feed Manufac. October 18-20, 2005. Syracuse, NY. Kononoff, P. J., A. J. Heinrichs, and D. R. Buckmaster. 2003. Modification of the Penn State Forage and Total Mixed Ration Particle Separator and the effects of moisture content on its measurements. J. Dairy Sci. 86:1858-1863. Lammers, B. P., D. R. Buckmaster, and A. J. Heinrichs. 1996. A simple method for the analysis of particle sizes of forage and total mixed rations. J. Dairy Sci. 79:922-928. Mertens, D.R. 1997. Creating a system for meeting the fiber requirements of dairy cows. J. Dairy Sci. 80:1463-1481. Mertens, D. R. 2002. Determination of starch in large particles. Ro-Tap shaker method. U.S. Dairy Forage Research Center, Madison, WI. Revised April, 2002. Minson, D. J. 1990. Forage in ruminant nutrition. Academic Press, Inc. New York, NY.

Poppi, D.P., R.E. Hendrickson, and D.J. Minson. 1985. The relative resistance to escape of leaf and stem particles from the rumen of cattle and sheep. J. Agric. Sci. (Cambridge) 105:9-14. Troelsen, J.E., and F.W. Bigsby. 1964. Artificial mastication - a new approach for predicting voluntary consumption by ruminants. J. Anim. Sci 23:1139-1142. Ulyatt, M. J. 1983. Plant fibre and regulation of digestion in the ruminant. Page 103 in Fibre in human and animal nutrition. Royal Soc. NZ, Wellington, NZ. Van Soest, P. J. 1994. Nutritional ecology of the ruminant. Cornell University Press. Ithaca, NY and London, UK. Welch, J. G., and A. P. Hooper. 1988. Ingestion of feed and water. pp 108-116 in The ruminant animal. Digestive Physiology and Nutrition. D. C. Church, ed. Prentice- Hall, Inc. Englewood Cliffs, New Jersey. Woodford, S. T., and M. R. Murphy. 1988. Dietary alteration of particle breakdown and passage from the rumen in lactating dairy cows. J. Dairy Sci. 71:687-696.