S-1000 Regional Project
Animal Manure and Waste Utilization, Treatment and Nuisance Avoidance for a Sustainable Agriculture
2003 Station Reports

Reporting Scientists from Purdue University
Objective 4, Task 1:

Reporting Scientists:    Todd Applegate

Objective:     Determine the effects of copper source on performance and phosphorus
        retention in broiler chicks.

Copper (Cu) is often added to broiler diets at therapeutic dosages due to its anti-microbial and growth promoting effects despite alleged chelation with and reduced utilization of phytate phosphorus (P).  Therefore male broiler chicks were fed diets containing 0 and 250 ppm Cu from Cu sulfate (SUL), Cu citrate (CIT), Cu lysinate (LYS) and Cu Chloride (CL) sources from 9 to 22 d of age (8 pens/treatment, 6 birds/pen).  Dietary concentrations of non-phytate P and calcium were formulated to be 0.2 and 0.7% respectively.  Body weight gain was not significantly different between treatments (P>0.05).  Supplementation with 250 ppm Cu from Cu LYS resulted in chicks having greater toe and tibia ash weights (P<0.001) as compared to chicks fed Cu SUL, but was not significantly different from birds fed the Cu CL, Cu CIT and 0 ppm Cu diets.  Supplementation with Cu LYS resulted in birds with a significantly greater  toe ash percentage (P<0.0001) as compared to Cu CIT, Cu SUL and the 0 ppm Cu diets, but was not significantly different than birds fed the Cu CL diet.  With respect to tibia ash percentage, birds fed the Cu LYS diet resulted in a significantly greater % tibia ash (P<0.0001) as compared to the Cu SUL and 0 ppm Cu diets, but was not significantly different than either the Cu CL or Cu CIT diets.  Supplementation with 250 ppm Cu SUL or Cu CIT reduced apparent P retention by 0.029 and  0.053 % of the diet, respectively (P≤0.05) as compared to 0 ppm diet; whereas the apparent P retention when 250 ppm Cu LYS or Cu CL was fed was not significantly different from the 0 ppm Cu diet (P>0.05).

Usefulness & Impacts:  Supplementation with 250 ppm Cu from either Cu CIT or Cu SUL resulted in decreased apparent P retention.  Supplementation with either 250 ppm Cu CL or Cu LYS, however, demonstrated improved apparent P retentions such that they were not different than the 0 ppm supplemental Cu diet.
Reporting Scientists:    Todd Applegate & R. Angel (U. MD)

Objective:     Determine the effect of dietary calcium on intestinal phytase activity and
        phytate-phosphorus utilization in Pekin ducklings.

Higher concentrations of calcium (Ca) in the diet may decrease phytate-phosphorus hydrolysis because of the chelation of Ca with the phytin molecule. Therefore, 192 drakes were fed 0.6, 0.8, 1.0, or 1.2 % Ca from 7 to 17 d of age (6 birds/pen and 8 pens/treatment).  Non-phytate phosphorus (nPP) was formulated to 0.4 %.  Excreta was collected from 15 to 17 days of age and the tibia and middle toe from each foot was collected on 18 d of age.  Mucosa was collected on 18 d of age from treatments with 0.6 and 1.2 % Ca for determination of intestinal phytase activity.  Body weight gain was greatest when the ducklings were fed the 1.0 % Ca diet and significantly different from that of the 0.6% Ca (P < 0.05), but not significantly different than birds fed the 0.8 and 1.2 % Ca diet.  Tibia ash percentage was not significantly affected by dietary Ca (P > 0.05).  Toe ash percentage, however, was greater in birds fed 0.8% Ca diet as compared to the other three diets (P< 0.0001).  Apparent phosphorus retention was not significantly affected by dietary Ca and averaged 46.35 % (P > 0.05).  Specific phytase activity within brush border vesicles prepared from intestinal mucosa and vesicle Ca concentration was not significantly affected by dietary Ca (P >0.05).  A positive correlation, however, was found between the Vmax and that of the Ca concentration within the vesicles (r=0.59, P< 0.02).

Usefulness & Impacts:  Maximal  duckling growth and toe ash percentage was realized when birds were fed 1.0 and 0.8 % Ca, respectively, which is significantly greater than the current 0.6 % Ca for ducks as reported by the National Research Council (1994).  Contrary to previous work in broilers, intestinal phytase was not significantly affected by dietary Ca concentration in ducklings. 
Reporting Scientists:    Todd Applegate & K. Roberson (Michigan State U.)

Objective:     Compare of Wheat Bran Phytase and a Commercially Available Phytase on
        Turkey Tom Performance and Litter Phosphorus Content

A 12 wk turkey study evaluated the effect of wheat bran phytase versus a commercially available phytase (Natuphos 600®) on growth performance, and the amount and forms of litter phosphorus (P).  At 5 wk of age, 780 Hybrid Converter turkey toms were randomly allocated into 32 floor pens with 23 to 26 birds/pen (avg. tom starting weight=1.74 kg).  A corn-soybean meal based mash diet was phase fed at 3 wk intervals.  Four dietary treatments were fed with 8 replicates.  Treatments consisted of T1: control (0.50, 0.44, 0.38, or 0.35% non-phytate P (npP) at 8, 11, 14, or 17 wk, respectively), T2: negative control (subtracting 0.1 percentage units of npP from T1 and 0.2 percentage units of Ca from T1 to maintain 2:1 Ca:npP), T3: Ca and P levels as T2 with 3.27% wheat bran (900 units/kg phytase activity), T4: Ca and P levels as T2 + Natuphos® (900 units/kg, 5-11 wk; 600 units/kg, 11-17 wk).  Measurements included BW, feed:gain (F:G), and litter P (soluble P, total P, soluble/total P).  T1 produced the greatest BW at all age periods (P<0.05).  At 8 and 14 wk of age, T3 and T4 had similar BW compared to T2. At 11 wk, T4 had higher BW than T2 (P<0.001).  At 17 wk, T3 had greater BW compared to T2 (15.27 vs. 14.87 kg; P<0.001), but not different from T4.  From 5-17 wk, T1 had improved F:G over T2 and T4 (P=0.012).  T2, T3 and T4 had similar F:G.  T4 litter soluble P was greater than T2 (T1: 0.19%; T2: 0.16%; T3: 0.18%; T4: 0.20%; P=0.046).  Total P was decreased by 26-28% when T3 or T4 was fed compared to T1 (P=0.006).

Usefulness & Impacts:  Feeding wheat bran phytase yielded similar growth, litter soluble P, total P and soluble P as a proportion of total P as Natuphos®.  Litter soluble P from Natuphos® was not different from control.
Reporting Scientists:    A. Sutton & B. Richert

Objective:     Determine the Effects of Reduced Crude Protein and Fiber Supplementation on Nitrogen and Phosphorus Utilization and Availability

    During this study the inclusion of wheat bran (WB) did decreased feed intake by 9%, however, the amount of manure produced (expressed in gal or lb) was not significantly different from the diets without WB.  Water usage was also not different among dietary treatments.  The amount of manure DM (lb/d) was increased with the addition of WB by 16.4%, while reduced CP levels decreased manure DM (ld/d) by 19.2%.  There was a trend for an interaction between CP and WB for manure DM production (lb/d), with WB having a greater effect on manure DM production (lb/d) from pigs fed the LNE diet compared to the control diet.  This may be explained by WB’s addition of fibrous and non-digestible fractions to the LNE diet, which was designed to reduce/minimize DM excretion compared to the control diet.  This can again be seen with a 32% increase in manure DM (%) with the addition of WB.  One would have also expected a change due to the fiber addition from the WB to enhance hindgut and manure storage fermentation.  However, there were no differences in manure pH among dietary treatments.  The lack of pH change is verified by the lack of change in VFA concentrations as well, which would have been expected with these fibrous additions.
    Otto et al. (2003) stated that by reducing dietary CP intake, reductions in N excreted would be possible.  This was evident in this trial, where total nitrogen concentration (DMB) in the stored manure column was decreased by 14.6% when CP was decreased.  This decrease in TN concentration was in fact due to the reduced soybean meal and amino acid content of the LNE treatments.  The amount of TN produced (expressed in g and g/d) was also affected by dietary modification of protein content, and decreased by approximately 32%, with the fibrous addition of the WB not affecting daily N excretion.
    Wheat bran’s ability to sequester AmmN was not observed in this study, with WB not increasing AmmN and TN in the stored manures.  Wheat bran actually decreased the concentration of AmmN (DMB) by 22.4% in the stored manures, while reducing dietary CP also decreased the amount of AmmN (g and g/d) by 27.7%.  The concentration and production of TN and AmmN in manure from pigs fed the control diet was higher, when the TN and AmmN should have been lower when compared to the control plus 10% WB diet.  However, the LNE diet does not follow the same pattern.  Manure from pigs fed the LNE plus 10% WB was higher in TN concentration on both an as-is basis and DMB when compared to the LNE diet. The AmmN (as-is) content of LNE plus 10% WB diet was numerically higher than the LNE diet, but not significantly different.
    Because of wheat bran’s natural phytase and fiber content a decrease in excreted phosphorus was expected due to the release of more phytic phosphorus.  Looking within each set of diets, the control plus 10% WB diet followed the trend of reducing P excretion compared to the control diet without WB.  On the other hand, the LNE plus 10% WB diet had a numerical increase in the excretion and concentration of total phosphorus. Reducing crude protein and dietary phosphorus with the LNE diets did significantly alter the concentration of TP (as-is and dry matter basis) in the store manure, decreasing by 38% and 39%, respectively.  The amount of TP produced (g and g/d) was also decreased by approximately 48.3%.  The decrease in concentration and excreted phosphorus was primarily a result of a 35.8% reduction in dietary phosphorus concentration with the LNE diets than the inclusion of WB. 

Usefulness and Impacts:  This study indicates that reductions in TN and AmmN concentrations are possible with the addition of 10% wheat bran in formulated diets.  This study also presents and supports the idea that reducing dietary crude protein and phosphorus levels in the diet causes a decrease in the amount of manure DM produced (lb/d), and a reduction in TN, ammonium N, and TP excreted in the manure.  
Reporting Scientists:    A. Sutton, S. Radcliffe, B. Harmon, & B. Richert

Objective:     Determine the Effects of Supplementing Weanling Pig Diets with Organic and Inorganic Acids on Growth Performance and Microbial Shedding

The objective of this study was to evaluate the performance of organic and inorganic acid blends and also a combination of these acids as potential alternatives to antibiotic growth promoters in nursery pig diets. While antibiotics help ensure a control of diarrhea outbreaks and mortality caused by bacterial infections in pigs, a risk of developing cross-resistance of pathogens to antibiotics used in human therapy has become more evident (Piva, 1998; Hillman, 2001 ). Due to this reason the swine industry is now searching for an equally competent, if not better, alternative to antibiotics in relation to growth performance and health in nursery pig diets.
Results from this study demonstrate that the overall growth performance of nursery pigs could be affected by the addition of growth promoting agents to their diets. It has been previously reported that the addition of antibiotic growth promoters to the nursery pig diet has significantly increased the growth performance over that of the control pigs.  In this study it was found that pigs fed a diet containing carbadox had the numerically the highest ADG, most favorable feed efficiency and the heaviest weights across treatments. However, during week 1 of the study the very opposite was found in this study, with the pigs being fed the carbadox diet having the poorest growth performance. This may be due to the abrupt change in diet of these nursery pigs and a poor acclimation period post-weaning by these pigs, as the growth performance following this first seven day period was highest for the pigs fed the carbadox diet.  The performance of pigs fed the organic and inorganic acid blend diets were similar to each other and somewhat intermediate between the carbadox-fed pigs and the pigs that received no growth promoting additives in their diets. The pigs fed the combination acid diet had the numerically the poorest growth performance, lower than the negative control. This may have been as a result of the high levels of acid in the diet (0.6%) suppressing feed intake and depressing overall performance.
    This study also demonstrated that the amount of E. coli being shed differs in relation to the dietary treatment being fed.  It was noted that the greatest reduction in E. coli shedding from d 6 to d 34 was seen in pigs fed the combination acid diet.  However, similar, less dramatic, reductions were also seen in pigs fed the inorganic and the organic acid blend diets. The amount being shed by the negative control pigs did not differ greatly throughout the entire study. E. coli counts for the remaining treatments were higher on day 6 of the study compared to the negative control, but by day 34 the E. coli counts for all the acid treatments were lower than for the pigs fed the negative or positive control diets. The presence of unidentified bacteria was also quantified during this study. Our results lead us to believe that there may be a competitive exclusion relationship existing between the E. coli and these unidentified bacteria. While the population of E. coli undergoes a decrease, the population of the unidentified bacteria increases.
The results of this study suggest that the presence of salmonella is not affected by these dietary treatments. Salmonella was consistently found in pigs fed the carbadox diet and also the organic acid blend diet throughout the study. This is most likely due to initial salmonella infection during the nursing period of lactation and was not eliminated during the post-weaning nursery period.
Overall, a fall in fecal pH took place during the course of the study. However, the fecal pH of pigs fed different dietary treatments were not statistically different from each other.

Usefulness & Impact  The use of organic acid and inorganic acid blends in nursery pig diets has been demonstrated in this study to have similar performance to the use of carbadox or no antibiotics. This study also shows that a combination acid diet may not be as beneficial, as the high acid content (0.6%) reduced feed intake and therefore growth performance of the nursery pig. The acid diets also reduced E. coli shedding compared with the medicated and non-medicated diets, which may have long term pig health benefits.
Reporting Scientists:    A. Sutton, B. Harmon, S. Radcliffe, A. Schinckel, & B. Richert

Objective:     Determine the Effect of Feeding a Reduced Crude Protein and Reduced
        Phosphorus Diet on Growth Performance and Carcass Characteristics in
        Grow-Finish Swine

Fifty barrows and forty-eight gilts (Danbred Hampshire X Duroc by Yorkshire X Landrace) were sorted by sex and weight, and randomly assigned to one of two dietary treatments, control and LNE (LNE = High available phosphorus corn + reduced crude protein, amino acid     supplementation + phytase). Feeding diets that incorporate reduced CP, amino acid supplementation, HAP corn, and phytase can maintain growth performance in grow-finish swine, providing similar performance to typical commercial control diets.  Pigs fed the LNE diets grew slower (P<0.05) and had slightly decreased feed efficiency during the grower phase. This slower growth rate and lower feed efficiency deficit was erased during the finisher phase, with the LNE pigs having very comparable growth performance data with that of the control pigs for the overall grower/finisher phase.
Carcass and ultrasound mass scan measurements for the ending grower phase and final slaughter yielded no differences (P>0.05) between the two treatments in backfat thickness, although, LNE pigs tended to have numerically greater backfat depths at the 10th rib, last rib, and last lumbar regions. Also, LNE fed pigs had LEAs that tended to be numerically smaller. Consequently, over the coarse of the scan period, predicted fat free lean % for the control pigs became greater (P<0.05) than that of the LNE pigs.
     It has been presented by Kendall et al. (1999) that the increase in backfat depths could be attributed an increase in the net energy of the LNE diets. The increase in net energy is due to the substitution of soybean meal with HAP corn in the LNE diets, which will lead to a more energy dense diet.  Shelton et al.(2003) reported that the edition of phytase also had a tendency to increase backfat depths, potentially due to liberation of some energy previously bound by the phytate molecule.

Usefulness & Impact  Diets formulated with reduced crude protein, crystalline amino-acid supplementation, high available phosphorus corn, and phytase, are able to meet the nutrient requirements of the pigs and maintain sufficient growth performance. Carcass quality was slightly negatively impacted with the LNE diet for reasons that are not fully understood at this time.
Reporting Scientists:    A. Sutton, S. Radcliffe, A. Schinckel, & B. Richert

Objective:     Determine The Effect of Feeding a Low Nutrient Excretion Diet on Growth
        Performance and Carcass Characteristics of Nursery, Grower, and Finisher
        Pigs

The purpose of this experiment was to combine these three feeding practices into the same low nutrient excretion (LNE = High available phosphorus corn + reduced crude protein, amino acid  supplementation + phytase).) diet and measure its effect on growth performance and carcass characteristics during the nursery, grower, and finisher phases of production.

Usefulness & Impact  The data from this study show that the feeding of low nutrient diets does not have significant detrimental impacts on pig growth performance or carcass characteristics other than a slight reduction in performance of pigs fed the LNE diet during the nursery and grower periods, but there is no difference in performance during the finisher and overall grow-finish periods. As in a previous trial at our research station, the feeding of LNE diets tends to lead to a numerical increase in backfat depth and a tendency for LEA to be smaller in the pigs fed the LNE diet. Ways to overcome these small differences between treatments could include: an evaluation of the net energy levels of the rations, adjust diets to more closely meeting the exact amino acid requirements of these pigs, or reexamine the suggested amino acid requirements and ratios for nursery and growing pigs that are given by the NRC (1998) for these time periods.
Reporting Scientists:    A. Sutton, S. Radcliffe, A. Schinckel, & B. Richert

Objective:     Determine the Effects of Low Phytic Acid Corn, Low Phytic Acid Soybean Meal and Phytase on Nutrient Excretion and Nutrient Digestibility in Pigs
B. Hill, S. L. Hankins, S. A. Trapp, A. L. Sutton, and B. T. Richert

Forty-eight barrows, 24 per replicate (initial body weight = 100 lbs) were blocked by weight and ancestry and randomly assigned to one of eight dietary treatments in a 2 x 2 x 2 factorial arrangement with all possible diet combinations of LPA corn, LPA soybean meal, normal (NRM) corn, NRM soybean meal, and supplemental phytase.  Total fecal DM excreted, %DM of feces, and %DM digested were not different among treatments in this experiment.  Furthermore, we saw no difference between treatments for as-is feces, urine, and total manure produced.  However, pigs consuming LPA soybean meal consumed less total DM than pigs fed NRM soybean meal (P < 0.03), 3.21 vs. 3.30 lb/d respectively.  In addition, pigs fed diets containing phytase consumed less DM per day (P < 0.016) than those without phytase inclusion, 3.20 vs. 3.30 g/d respectively.  Pigs fed LPA corn in diets had a higher overall fecal pH (P < 0.047) than those fed NRM corn, 5.7 vs. 5.6.
Nitrogen digestibility, fecal N, urinary N, and total N excreted (Table 3) were not significantly different between treatments.  However, N absorbed was significantly higher (P < 0.0003) for diets containing no phytase than for those with phytase inclusion, 46.9 vs. 43.3 g/d respectively.  In addition, N retained was higher for pigs fed diets without phytase (P < 0.009) compared to those diets with phytase inclusion, 26.3 vs. 23.5 g/d respectively. 
Ammonium N excreted in the feces was significantly higher (P < 0.005) for pigs fed diets containing LPA soybean meal compared to NRM soybean meal, 1.90 vs. 1.53 g/d respectively.
Fecal phosphorus excretion  was reduced 10% for pigs fed LPA corn vs. NRM corn, 2.87 vs. 3.22 g/d (P < 0.05), 17% for pigs fed LPA soybean meal vs. NRM soybean meal, 2.74 vs. 3.34 g/d (P < 0.001), and 18% for pigs fed phytase vs. non-phytase diets, 2.74 vs. 3.35 g/d (P < 0.02).  In addition to these main effects, there were additive benefits of reduced P excretion which were a 28% reduction for pigs fed LPA corn and LPA soybean meal vs. NRM corn and NRM soybean meal, 2.51 vs. 3.47 g/d (P < 0.0001), and a 43% reduction for pigs fed LPA corn, LPA soybean and phytase vs. NRM corn and NRM soybean meal without phytase, 2.13 vs. 3.76 g/d (P < 0.0001).  No significant differences were detected among treatments for urinary P excretion.  Total P excretion values, therefore, correspond to fecal P excretion numbers. 

Usefulness & Impact  Feeding of any combination of LPA corn, LPA soybean meal, and phytase can significantly improve P digestibility while dramatically decreasing P excretion.  and thus reduce the potential negative impacts of swine production on the environment.
Planned research 2003-2004
 Peer reviewed Publications issued 2002-2003

Angel, R., N.M. Tamim, T.J. Applegate, A.S. Dhandu, L.E. Ellestad, 2002.  Phytic acid chemistry:  influence on phytin-phosphorus availability and phytase efficacy.  J. Applied Poultry Res.11:471-480.

Angel, R., T.J. Applegate, and S. Bastyr.  2003.  Biological mass balance versus model based on mass balance approach.  Intl. Symp. Animal, Ag. Food Proc. Wastes (in press).

Applegate, T.J., L.P.V. Potturi, and R. Angel.  2003.  Model for estimating poultry manure nutrient excretion:  a mass balance approach.  Intl. Symp. Animal, Ag. Food Proc. Wastes (in press).

Applegate, T.J., D.M. Webel, and X.G. Lei.  2003.  Efficacy of E. coli Phytase expressed in yeast on phosphorus utilization and bone mineralization in turkey poults.  Poultry Sci. (in press).

Applegate, T.J., R. Angel, and H.L. Classen, 2003.  Effect of dietary calcium, 25-hydroxycholecalciferol, and bird strain on small intestinal phytase activity in broiler chickens.  Poultry Sci. 82:1140-1148.

Applegate, T.J., B.C. Joern, D. L. Nussbaum-Wagler, and R. Angel, 2003.  Water soluble phosphorus in fresh broiler litter is dependent upon phosphorus concentration fed but not on fungal phytase supplementation.  Poultry Sci.  82:1024-1029.
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