abstracts of papers seeds, etc). Relatively high variability was noted in six bakery by-product samples (starch, 24.7-46.5%; sugars, 5.8-10.5%; fat, 6.5-10.2%; NSP, 3.417.0%; phytate P, 0.28-0.47%). Three samples of mill run were evaluated and were found to contain 16.0-18.2% protein, 26.0-30.4% starch, 3.24.0% sugars, 3.7-5.8% fat, 20.9-22.4% NSP and 0.63-0.83% phytate P. Samples selected for in vivo study included wheat screenings (3), bakery by-products (2) and mill run (1). Broiler chicken performance (2 week growth trial) was compared using control (wheat/soybean meal based diet) and diets substituted with wheat by-products (wheat screenings, 20%; bakery by-products or mill run, 10%). Broiler chicken performance was not affected by dietary substitution of wheat by-products for wheat as values for control, wheat screenings, bakery by-products and mill run were similar for body weight gain (514, 520, 515, 500 g/bird/14 days) and for feed to gain ratio (1.52, 1.52, 1.52, 1.50), respectively. Response to enzyme supplementation (xylanase, β -glucanase and a broad spectrum of other enzyme activities) was highest for wheat screenings showing a statistically significant (P≤0.05) improvement in body weight gain (523 vs. 548 g/bird/14 days) and feed to gain ratio (1.54 vs. 1.47). There was a week relationship between intestinal viscosity, enzyme supplementation and broiler chicken performance. It is evident from this study that the use of wheat by-products in concert with an effective enzyme supplement will allow for cost effective formulation of poultry diets. Key Words: Wheat by-products, Chemical compostition, Nutritive value, Broiler chicken, Enzyme 86 rA blend of xylanase, amylase and protease (Avizyme 1500) improves laying hen egg production, feed efficiency, and livability when supplemented in corn-soy based diets. M.E. Cook*1 , B.M. Drake1 , and E.E.M. Pierson, 1 Department of Animal Sciences, University of Wisconsin, Madison, WI, 2 Finnfeeds (A Division of Danisco Cultor), Fenton, Missouri. Avizyme 1500 is a feed grade enzyme blend containing xylanase, amylase, and protease, specifically designed for diets formulated with corn/sorghum and soybean meal. Research with broilers has shown that its dietary use improves the digestibility of protein, starch and fat, and improves weight gain and feed efficiency. The objective of this trial was to determine if dietary Avizyme 1500 would improve egg production and feed efficiency in laying hens. Pullets (Bovans strain, 18 wks of age) were housed 3 to a cage with 10 cages per replication. Five replications were assigned to each of 6 dietary treatments analyzed in a 2 (with or without .075% enzyme blend) by 3 (diet type) factorial arrangement of treatments. The diet types consisted of: 1.) Standard commercial formula (Com): 2.) Diet 1 plus a 3% increase in dietary methionine and lysine (Spec Up): 3.) Diet 1 with 3% less metabolizable energy (Spec Down). Feeding of test diets commenced with light stimulation (increased from 12-16 hr over 8 wks). Test diet nutrient levels were adjusted according to stage of production. Egg production, mortality, and feed efficiency were monitored for six 28 day treatment periods. Results presented are cumulative: livability (cliv); eggs per hen house (cehh), feed per hen (cfeed), feed per dozen eggs (cf/doz). No enzyme by diet interaction was observed, hence all significant differences were attributed to enzyme use or diet type. Beginning with period 2 throughout the study, Avizyme 1500 significantly improved cliv (3-4%) and reduced cf/doz (3.5-4%) with no effect on cfeed. Cehh was increased (2.7-6%) with Avizyme 1500 in period 2-4, but was not significantly different in period 5 (p<.07) or 6 (p<.09). Throughout the study, hens fed the Spec Down diet consumed more cfeed, but had similar cehh and cf/doz compared to Com. Both the Spec Down and Com diets had reduced cehh and cf/doz compared to the Spec Up diet. Over the 6 feeding periods, hens fed Avizyme 1500 had a return above enzyme cost of $.135 per hen (assuming feed costs at $85 US/ton, egg value at $.50/dozen and Avizyme value at $1.695 US/ton). Economic improvements related to Avizyme supplementation occurred regardless of diet type but the greatest effects were seen when feeding the Com and Spec Up diets. Key Words: Avizyme, Enzyme, Egg production, Feed efficiency, Livability 87 Influence of microbial phytase on bone ash and performance of broilers. P. Cachaldora1 , M. J. Aranibar2 , J. Mendez1 , and G. G. Mateos*2 , 1 Coren SCL, Spain, 2 Universidad Politecnica de Madrid, Spain. An experiment was conducted to evaluate the influence of phytase supr plementation (Natuphos 5000) on strength and ash content of tarsus and performance of broiler chicks. A completely randomized design was 21 used with four treatments arranged as a 2x2 factorial with two levels of available phosphorus (0.40 vs 0.32% from 0 to 21 d and 0.37 vs 0.29% from 22 to 42d of age) and two levels of phytase supplementation (0 vs 500 FTU microbial phytase/kg). There were six replicates per treatment (70 straight run chicks per pen). The diets were based on wheat, corn, soybean meal and meat and bone meal with 3,010 and 3,150 kcal ME/kg and 1.36 and 1.24% total lysine from 0 to 21 and from 21 to 42d, respectively. At the end of the trial four birds of each pen were killed by cervical dislocation and the tarsus was removed for measuring bone strength and ash content. At 42d of age no differences among treatments were detected for average daily gain (57.7, 57.0, 57.3, and 57.7 g/d, respectively; P>0.05), or feed conversion (1.71, 1.72, 1.72, and 1.71 g/g, respectively; P>0.05). Neither available phosphorus level nor phytase supplementation influenced tarsus strength (13.8 vs 14.6 kg/mm2 and 14.4 vs 14.0 kg/mm2 , respectively) but phytase supplementation tended to improved tarsus ash content (31.8 vs 32.7%, P=0.09). We conclude that phytase supplementation improved bone ash and that 500 FTU of phytase are equivalent to at least 0.08 % of available phosphorus in commercial diets for broilers. Key Words: Broiler, Available phosphorous, Microbial phytase 88 Effect of dietary calcium concentration and 25hydroxycholecalciferol on phytate hydrolysis and intestinal phytase activity in broilers. T.J. Applegate*1 , R. Angel1 , H.L. Classen2 , R.W. Newkirk2 , and D.D. Maenz2 , 1 Department of Animal and Avian Sciences, University of Maryland, College Park, 2 Department of Animal and Poultry Sciences, University of Saskatchewan. Broilers have a limited ability to utilize phytate phosphorus (PP) and the influence of nutrients on the activity or efficacy of intestinal phytase is relatively unclear. Therefore in a 2x2 factorial experiment, Hubbard-Peterson male broilers were fed diets with or without 25hydroxycholecalciferol supplementation (25-OH D3 ; 0.21 mg/kg) with varying dietary calcium (Ca) concentrations (0.4 and 0.9%) from 7 to 21 d of age (5 pens/treatment, 3 birds/pen). All diets were formulated to contain 2200 IU vitamin D3 /kg diet. Diets were analyzed to contain 0.30% PP (0.51% total phosphorus). Neither dietary Ca concentration nor 25-OH D3 supplementation significantly affected broiler performance (P > 0.05). Phytase activity within brush border vesicles prepared from small intestinal mucosa (Vmax , nmol P released/mg protein/min) was 1.6 times greater in broilers fed 0.9% vs 0.4% Ca (P ≤ 0.05). Intestinal phytase activity was not significantly affected by 25-OH D3 supplementation (P > 0.05). The additional intestinal phytase activity in broilers fed 0.9% vs 0.4% Ca did not significantly affect apparent ileal PP hydrolysis which averaged 53.4% (P > 0.05). Dietary supplementation with 25-OH D3 increased apparent ileal PP hydrolysis from 42.9 to 64.0% when compared with broilers that were not supplemented with 25-OH D3 (P ≤ 0.05). In conclusion, high dietary Ca (0.9%) increased intestinal phytase activity but was not effective in increasing PP hydrolysis due presumably to the binding of Ca with the phytate molecule. Whether 25-OH D3 supplementation increased PP hydrolysis by increasing intestinal Ca or P absorption is uncertain. Nevertheless, the activity of intestinal phytase was enough to facilitate this hydrolysis. Key Words: Calcium, 25-Hydroxycholecalciferol, Intestine, Phytase, Phytate 89 Effect of dietary non-phytate phosphorus (nPP) on performance and bone measurements in broilers fed on a four-phase feeding system. R. Angel*, T.J. Applegate, and M. Christman, Department of Animal and Avian Sciences, University of Maryland, College Park. Three experiments were done to determine the effects of nPP, on performance and bone measurements in male (Ross 308) broilers, independently in each of three phases of a four-phase feeding system. Hatchling were raised to 18, 32 or 42 d of age on starter (ST), grower (GR), and finisher (FN) diets that met NRC (1994) recommended nutrient levels. In each of the GR (18 to 32 d), FN (32 to 42 d) and withdrawal (WD, 42 to 49 d) phases a 2 x 4 factorial arrangement of two calcium (Ca) and four nPP levels with eight floor pens (60 birds per pen) per treatment were used. The analyzed Ca and nPP (total phosphorus (P) minus phytate P) concentrations were: Gr phase, 0.88, 0.80% Ca and 0.45, 0.36, 0.32, 0.28% nPP; FN phase, 0.80 and 0.70% Ca and 0.34, 0.28, 0.24, 0.19% nPP; and WD phase, 0.70 and 0.61% Ca and 0.25, 0.20, 0.17, 0.11%nPP. At the end of each experiment, four birds of average pen weight were taken and placed in battery cages for three days. After three days excreta were 22 abstracts of papers collected (total) and right tibias sampled for breaking strength and ash (TA, dry-defatted basis) determinations. In the Gr phase, there was an effect (P<0.05) of Ca (positive effect of low Ca) and a quadratic (P<0.05) effect of nPP on BW, BW gain, and feed efficiency (F/G) and a Ca by nPP interaction (P<0.05) on BWG. In the quadratic effect of nPP, the inflection point is between 0.32 and 0.28% nPP. In the FN phase, level of nPP had quadratic (P<0.01) effect on BW gain and F/G but no Ca by nPP (P>0.05) interaction were seen. No (P>0.05) Ca, nPP or Ca by nPP interaction effects were seen on any performance measures in the WD phase. Tibia ash was quadraticaly (P<0.05) affected by nPP but no (P>0.05) Ca or Ca by nPP interactions. In the quadratic effect of nPP, the inflection point is between 0.24 and 0.19% nPP. In the FN phase, nPP level had quadratic effect (P<0.005) on TA but no Ca or Ca by nPP interaction was seen. In the WD phase, the was no effect (P<0.05) of nPP or Ca on TA. Key Words: Non-phytate phosphorus, Bone, Broilers, Requirement 90 Effect of dietary non-phytate phosphorus (nPP) level on broiler performance and bone measurements in the starter and grower phase. R. Angel*1 , T.J. Applegate1 , M. Christman1 , and A.D. Mitchell2 , 1 Department of Animal and Avian Sciences, University of Maryland, College Park, 2 USDA/ARS, Beltsville, Maryland. An experiment was conducted to determine nPP requirements of male (Ross 308) broilers in the starter (ST, hatch to 17 d) and grower (GR, 17 to 31 d) phases. An incomplete 2 x 3 factorial design was used with two nPP levels (0.43 and 0.35%) in the ST and three nPP levels in the GR phase. Calcium (Ca) was 0.9 and 0.81% in the ST and GR phases, respectively. Forty battery pens (eight birds per pen) were assigned to five treatments such that broilers fed 0.43% nPP in the ST phase were randomly assigned to three GR nPP treatments (0.36, 0.31, and 0.26%) and those fed 0.35% nPP (ST phase) were randomly assigned to two nPP treatments (0.31, and 0.26%) in the GR phase. Right tibia and femur, and middle toe from both feet were collected from three birds per pen at 17 and 31 d for bone measurements. No effect (P>0.05) of nPP level was seen in the ST phase on performance and only gain was affected (P<0.05) by nPP level in the GR phase. In the ST phase, tibia epiphyseal (upper and lower 25% of bone) ash and tibia shaft (middle 50% of bone) ash (dry defatted basis) were higher (P<0.05) for birds fed the 0.43 vs 0.35% nPP. Level of nPP had no effect (P>0.05) at 17 d on tibia strength measures (stress, strain, modulus of elasticity, and breaking strength). Similarly, no differences in strength measures of the femur at 17 d were seen except for a decrease (P<0.05) in stress and breaking strength in birds fed 0.35 vs 0.43% nPP. At 31 d, nPP had an effect (P<0.05) on tibia epiphyseal and shaft ash. Epiphyseal tibia ash was lower (P<0.05) in birds fed 0.26 vs 0.31% nPP (42.8 vs 45.4%, SEM of 0.56%, respectively) at 31 d. Similarly, tibia shaft ash was lower (P<0.05) in birds fed 0.26 vs 0.31% nPP (52.2 vs 53.9%, SEM of 0.36%, respectively). Total body bone mineral density (BMD) as determined by dual energy x-ray absorptiometry was lower in birds fed the 0.26 vs 0.31% nPP (0.342 vs 0.355 gr/cm2 , SEM of 0.004 for BMD, respectively). Key Words: Non-phytate phosphorus, Requirements, Broiler, Bone 91 The effect of phytase utilization on production and manure parameters of hens of different ages. D. R. Sloan*, K. M. Gattas, and D. P. Eberst, University of Florida, Gainesville, FL, USA. Two experiments were conducted utilizing Hy-Line W-36 hens. In Experiment 1, two levels of calcium (3.05 and 3.50%) and three levels of added phosphorus (0.0, 0.1 and 0.2%) were fed with and without phytase to six replicate pens of four hens per pen from 46 to 58 weeks of age. In Experiment 2, hens were fed 3.50% calcium and either 0.08, 0.11, 0.14, 0.17, or 0.20% added inorganic phosphorus either with or without phytase from 61 to 70 weeks of age. There were five replicate pens of four hens per replicate fed each of the ten experimental diets. In Experiment 2, hens fed the unsupplement phosphorus diets without phytase, regardless of calcium level decreased feed intake within two weeks. Egg production was significantly reduced during week four and remained low throughout the remainder of the study. The addition of phytase on 0.10% inorganic phosphorus returned production levels to normal. Egg weights were significantly reduced by the lack of phosphorus supplementation at both calcium levels during week three. Specific gravity was likewise affected during week two. Manure phosphorus levels were significantly decreased at the 0 and 0.1% added phosphorus levels with and without phytase. In Experiment 2, all production parameters and manure phosphorus levels were unaffected by organic phosphorus or phytase addition. Key Words: phosphorus level, phytase, production, manure phosphorus 92 Effects of low-phytate corn and lowoligosaccharide, low-phytate soybean meal on performance, bone traits and phosphorus excretion by growing chicks. G.L. Cromwell*1 , E.G. Xavier1 , L.W.O. Souza1 , M.D. Lindemann1 , T.E. Sauber2 , H.L. Stilborn2 , and D.W. Rice2 , 1 University of Kentucky, Lexington, 2 Optimum Quality Grains, Des Moines, IA. Diets containing low-phytate corn (LP-corn) or near-isogenic normal corn (N-corn) and low-oligosaccharide, low-phytate soybean meal (LP-SBM) or normal soybean meal (N-SBM) from near-isogenic soybeans were evaluated. The LP-corn, N-corn, LP-SBM, and N-SBM contained .28, .25, .77, and .70% total P; .10, .20, .22, and .48% phytate P; and .18, .05, .55, and .22% non-phytate P; with estimated P bioavailabilities of 50, 10, 58, and 28%, respectively. Male, broiler-type chicks (n = 315) were allotted at 3 d of age to nine treatments (five pens of seven chicks/pen, 70 g BW). Diets 1-3 consisted of N-corn and N-SBM formulated to contain .45, .35, and .25% non-phytate P. Diets 4-6 were N-corn and LP-SBM containing .45, .35, and .25% non-phytate P. Diets 7-9 were LP-corn and LP-SBM containing .45, .37, and .29% non-phytate P. Monocalcium phosphate and calcium carbonate were used as sources of supplemental P and Ca. Diet 9 contained no added P. Total P levels were .72, .62, .53; .64, .54, .45; and .58, .50, .42%, respectively. All diets contained 1.2% lysine, .93% S-amino acids, 1.0% Ca, and 3,282 kcal TME/kg. LP-SBM was assumed to be 10% higher in TME based on its lower oligosaccharide (1.1 vs 6.3%) and higher sucrose (12.3 vs 7.2%) content. Cr2 O3 was included in all diets to estimate P excretion at 7 and 14 d. At the end of the 14-d experiment, all chicks were killed and tibias were removed. Gains and feed:gains were: 438, 406, 351, 416, 391, 343, 409, 400, 351 g; 1.28, 1.31, 1.35, 1.32, 1.29, 1.37, 1.28, 1.31, 1.35 for chicks fed Diets 1-9, respectively. Gain, feed:gain, and tibia strength and ash improved (P < .01) with increasing level of dietary P. Chicks fed LP-SBM excreted less P in their excreta, especially when LP-SBM was combined with LPcorn. The results indicate that chicks fed LP-corn and LP-SBM require less supplemental P and excrete less P than chicks fed typical corn-SBM diets. Key Words: Chick, Soybean Meal, Phosphorus, Phytate Saturday, AM, PATHOLOGY 93 Effects of F-strain Mycoplasma gallisepticum on blood characteristics in commercial laying hens. M.R. Burnham*1 , E.D. Peebles1 , S. Whitmarsh1 , and S.L. Branton2 , 1 Mississippi State University, 2 USDA,ARS, South Central Poultry Research Lab, Mississippi State, MS. The effects of F-strain Mycoplasma gallisepticum (FMG) on blood characteristics and nutrient absorption in commercial laying hens were investigated through 54 wk of age (34 wk of lay). Ten hens were assigned to each of eight negative pressure biological isolation units. Birds in four units served as uninoculated controls and those in four other units were inoculated with FMG at 12 wk of age. Blood variables tested were hematocrit (HCT), serum cholesterol (SCH), triglycerides (ST), high density lipoprotein cholesterol (HDLC), calcium (SC), and total plasma protein (TP). Eggshell conductance (EC) was also examined. Feed and fecal analysis were performed at 26 and 54 wk for determination of moisture, ash, CP, crude fat (CF), crude fiber (CFI), total dietary nutrients (TDN), nitrogen, and ME. Main effects due to bird age were observed for HCT, HDLC, SC, EC, and fecal CF, CFI, TDN, ME, and ash. Bird age by treatment interactions approached significance (P≤0.08) for SCH, ST, TP, and fecal moisture. These data suggest that FMG inoculation at 12 abstracts of papers wk may have an influence on SCH, ST, TP, and dietary moisture uptake in commercial laying hens, and may contribute to egg production shifts at 45 wk of age. Key Words: Blood, Layers, Mycoplasma gallisepticum, Triglycerides, Protein 94 Effects of an immunomodulator and phasic challenges of S6 strain Mycoplasma gallisepticum and Escherichia coli on egg production and egg characteristics of commercial laying hens during pre-peak lay. M.S. Jones*1 , S.L. Branton2 , E.D. Peebles1 , K.O. Willeford3 , T. Pharr4 , R. Montgomery4 , T.A. Parker3 , S. Whitmarsh1 , and W. Maslin4 , 1 Poultry Science Dept., Mississippi State University, 2 USDA, ARS, SCPRL, Mississippi State, MS 39762 , 3 Biochemistry & Molecular Biology, Mississippi State University, 4 College of Veterinary Medicine, Mississippi State University. The effects of S6 strain Mycoplasma galisepticum (MG) challenge and dual challenges of MG and Escherichia coli (serotype O-35) with or without an immunomodulator (derived from caprine serum) on egg production and egg characteristics in commercial laying hens were investigated through 26 wk of age. Ten hens were randomly assigned to each of 16 negative pressure biological isolation units. The four treatment groups consisted of: negative control, pre-lay challenge with MG (10 wk), dual challenge with MG (20 wk) and E.coli (22 wk), and Caprine factor (CF) treatment in conjunction with dual challenges of MG (20 wk) and E. coli (22 wk). CF was injected on Days 0 and 5 of each challenge. Weekly means for egg production (EP), egg weight (EW), Haugh units, blood/meat spots, and eggshell breaking strength were compared. Mean BW, eggshell weight per unit surface area (SWUSA), and percentages of egg yolk, albumen, and eggshell weight were compared biweekly. None of the variables examined were affected by treatment; however, BW, EP, EW, relative yolk weight, Haugh units, and eggshell breaking strength were affected by bird age. The performance of commerical laying hens during pre-peak lay were not significantly affected by individual challenge of MG or dual challenge of S6 strain MG and E.coli (serotype O-35) with or without a caprine serum immunomodulator. Key Words: Caprine factor, Escherichia coli, Egg production, Layers, Mycoplasma gallisepticum 95 Development of cultivation techniques and a sensitive PCR for duck hepatitis B virus. Chi-young Wang*1 , Joseph J. Giambrone1 , and Lois B. Allen, 1 Auburn University. Hepadnaviridae are a family of DNA-containg viruses that include human hepatitis B virus (HBV), woodchuck hepatitis virus and duck hepatitis B virus. Human hepatitis B virus is a worldwide important public health problem with several hundred carriers being at high risk of developing cirrhosis and hepatocarcinoma. Thus far,that exists neither a good cell culture system or any laboratory animals that allow the potential testing of several drugs or disinfectants. Duck hepatitis B virus is very similar to human hepatitis B virus in its sensitivity to drugs and disinfectants. Therefore, it has already been widely accepted as the best surrogate for human hepatitis B virus study. In this study, we will attempt to establish different cultivation techniques using duck hepatocytes, duck embryonic hepatocytes, and freeze-thawed hepatocytes. A sensitive PCR assay will be used to determine the titer of duck hepatitis B virus from infected duck serum and different culture cells. Through our study,duck hepatitis B virus could readily propagate and get the highest titer in the duck embryonic hepatocytes. This system will be successfully applied in a wide rande of study in the future. Key Words: Duck hepatitis B virus, Human Hepatitis B virus, Polymerase Chain Reaction 96 Minimum infectious dose of avian pneumovirus in turkeys. Faris F. Jirjis*, Sally L. Noll, David A. Halvorson, Kakambi V. Nagaraja, and Daniel P. Shaw, University of Minnesota, St. Paul, MN, USA. The aim of this study was to determine the minimum infectious dose of avian pneumovirus (APV) that would cause the disease and induce a serological response. Five groups of 4-week-old poults shown to be free of APV antibody were inoculated with 10-fold serial dilutions of cell culture propagated virus. Turkeys in each group received 50 µl in each nostril and conjunctival space of inoculum containing 101.3 , 102.3 , 103.3 , 104.3 , or 105.3 tissue culture infective dose 50 (TCID50 ) of avian 23 pneumovirus per ml. The poults were examined daily for clinical signs. At 5 days post-inoculation (PI), swab samples were collected from the choanal cleft and trachea for reverse transcriptase-polymerase chain reaction and virus isolation. Blood was collected at 7 and 14 days PI for APV by enzyme linked immunosorbent assay. In the groups given 101.3 and 102.3 TCID50 /ml, no evidence of infection was noted. Respiratory signs were seen between 2 and 12 days PI in groups that received 103.3 , 104.3 , and 105.3 TCID50 /ml. These groups developed serological response at 2 weeks PI and APV was detected from the choanal swabs at 5 days PI. The minimum dose that caused clinical signs and induced serological response was 103.3 TCID50 /ml. Key Words: Turkeys, Avian pneumovirus, Respiratory disease 97 Colonization studies of Campylobacter jejuni and Salmonella enteritidis in the intestinal tract of chickens. D.C. Wilkie* and A. Estrada, University of Saskatchewan. Campylobacter jejuni and Salmonella enteritidis are the most prevalent foodborne human pathogens associated with poultry meat. In spite of their recognition as causative agents of disease, and their persistence in poultry flocks, work to date has not resulted in their elimination from the food supply. The successful control of these microorganisms requires a comprehensive understanding of the ability of these microorganisms to become established in the intestine of chickens, and the factors which contribute to their ability to proliferate. However, very few studies have been conducted examining their intestinal colonization patterns. To better characterize these microorganisms in the gut environment, and to examine their role in the overall microbial ecology of the gut, fluorescent microscopy was used to study intestinal colonization by Campylobacter jejuni and Salmonella enteritidis. Intestinal cryostat sections of experimentally infected birds were examined by epifluorescent microscopy (Epi) and Scanning Confocal Laser Microscopy (SCLM) using species specific, affinity chromatography-purified fluorescent antibody probes. At days 4, 8, and 11 post-infection, both Epi and SCLM revealed a gradient of epithelial surface association in the intestinal tract of the infected birds. The highest degree of colonization was observed in the ceca, followed by the ileum, and lastly the jejunum. Campylobacter jejuni and Salmonella enteritidis colony plate counts obtained from intestinal contents confirmed the spatial population trends observed by microscopy at the different sampling times. These observations constitute the background for future investigations aimed at the study of diverse strategies for the reduction of these pathogens in broiler flocks. Key Words: Campylobacter jejuni, Salmonella enteritidis, Intestinal colonization 98 Ovotransferrin, a clinical marker of poultry inflammatory disease. H. Xie*1,2 , N. C. Rath1 , G. R. Huff1 , J. M. 1 Balog , and W. E. Huff1 , 1 USDA-ARS-PPPSR, 2 Dept. of Poultry Science, Univ. of Arkansas. Inflammatory agents of both biotic and abiotic origins induce elevation of certain serum proteins, which have potential as nonspecific clinical markers of health problems. These serum proteins, called ”acute phase proteins”, have been little understood in poultry. Our previous studies on broiler chickens injected with bacterial endotoxin showed an increase in serum levels of a protein with an approximate MW of 65 kDa. To determine whether this protein was elevated in other diseases, we compared serum protein profiles of chickens and turkeys with different health problems as well as under experimental conditions where the birds were injected with endotoxin, dried Staphylococcus aureus cells, Escherichia coli, reovirus, Eimeria tenella, and croton oil. Chickens with metabolic disorders such as ascites and tibial dyschondroplasia (TD) were also included. The results showed that 65-kDa protein was differentially elevated in sera from both chickens and turkeys with Escherichia coli and reoviral infections, and with administration of endotoxin, dried Staphylococcus aureus cells, and croton oil but not in metabolic diseases such as ascites and TD. We isolated this 65-kDa protein using 2-dimensional SDS-PAGE and conducted a N-terminal sequence analysis. A BLAST sequence similarity search in GenBank showed high homology with ovotransferrin, a major protein in egg white. Using commercial ovotransferrin as standard, this 65-kDa protein was tentatively identified as ovotransferrin. Even though most of the birds used in our studies were male and sexually immature, the results suggest that ovotransferrin may be a normal serum component in poultry, which is elevated during inflammation. Ovotransferrin is an iron transport protein with bactericidal activity. During infection and inflammation, ovotransferrin may play a 24 abstracts of papers role in physiological homeostasis and may be a useful indicator for tissue injuries caused by various inflammatory diseases. wheat middling administration compares with feed removal with regards to second cycle egg production remains to be investigated. Key Words: Serum protein, Ovotransferrin, Poultry, Inflammation Key Words: Molting, Salmonella enteritidis, Stress, Egg lay 99 Efficacy of an IBDV immune complex vaccine administered in ovo to 18 day old embryos with different levels of maternal antibody. M.M. Corley*, J.J. Giambrone, and T.V. Dormitorio, Auburn University, Auburn Alabama. 102 Comparative identification of microorganisms from commercial poultry operations by API20E and BBL Crystal Enteric/Nonfermenter systems. Y. Vizzier-Thaxton*1 , J.D. Tankson1 , J.H. Faison2 , and A.R. Messersmith2 , 1 Mississippi State University, Mississippi State, MS, 2 Marshall Durbin Science & Technology Center, Jackson, MS. The efficacy of an immune complex infectious bursal disease (IBDV) vaccine (2512-ICX), administered in ovo to embryos carrying different levels of maternal antibody (MatAb) was investigated. Embryos were obtained from a 29 wk old (high MatAb) and a 63 wk old (low MatAb) flock. Embryos at 18 days of age were vaccinated with 100 EID50 of either IBDV2512 or 2512-ICX. On days 3,6,9,15, and 21 post vaccination (PV), birds were sacrificed and bursa, spleen and thymus collected. Challenge studies were conducted on the progeny of the 29 wk old flock to determine the efficacy of the vaccines. To determine the optimum point of viral replication, tissue homogenates were prepared and vaccine viral particles were released by freon treatment and centrifugation at 12,000 X G for 30 minutes. A newly developed antigen capture chemiluminescent ELISA assay was conducted on the aqueous phase of the homogenate. This procedure allowed detection of IBDV in tissues weighing as little as 0.1g. Either vaccine was detected in 1-day-old chickens (3-days PV). The IBDV vaccine virus migration was traced from the bursa to the spleen, and subsequently to the thymus. The 2512-ICX vaccine virus replication peaked at day 6 PV and started to diminish on day 9 PV. Challenge studies indicated that the ICX vaccine protected 3 wk old birds with high levels of maternal antibody. Key Words: Chemiluminescence, ELISA, IBDV 100 Studies on the pathogenicity of infectious bursal disease virus associated with proventriculitis. T.V. Dormitorio*1 , J.J. Giambrone1 , and F.J. Hoerr2 , 1 Auburn University, Auburn Alabama, 2 Alabama State Veterinary Diagnostic Laboratory, Auburn, AL. Proventriculitis along with immunosuppression has recently been a problem of grow-out farms in North Alabama. Proventriculus tissues of sick birds aged 3-7 weeks from six different farms were homogenized and used to challenge 3-week old specific pathogen free (SPF) birds. After 7 and 14 days post infection, birds were sacrificed and necropsied. Tissue samples such as bursa, spleen and proventriculus were collected for histological analysis and IBDV isolation by PCR. Three groups of birds inoculated with 2054, 494A, and 707B proventricular homogenates showed consistent findings including enlarged proventriculi and atrophied bursae. Histological examination confirmed proventriculitis. By using nested RTPCR, IBDV was also isolated from these tissues. The PCR products are currently being sequenced and will be analysed to determine the molecular characteristics of these IBDV strains associated with proventriculitis. Key Words: IBDV, Proventriculitis, Nested RT-PCR, Immunosuppression 101 Comparison of Salmonella enteritidis levels in hens subjected to different molt procedures. K.-H. Seo, P.S. Holt*, and R.K. Gast, USDA-ARS Southeast Poultry Research Laboratory. Molting is an important economic tool for the layer industry as a means to maximize the effective laying life of a flock. Previous work showed that molting birds through feed removal increased the severity of a Salmonell enteritidis (SE) infection. We conducted a study to follow the progression of an SE infection in unmolted hens vs hens molted via 14-day feed removal or through ad libitum feeding of wheat middlings, a low-nutrition by-product of wheat processing. All hens were infected with 5 X 106 SE at day 3 of molt and sampled for SE shedding on days 4 and 11 post infection and SE organ levels at day 7. Both molt procedures caused cessation of egg lay within 5-6 days. In both trials, birds subjected to total feed removal shed 3-5 logs more SE than either the cotnrol birds or birds fed wheat middlings on the two sampling days. Liver/spleen, ovary, and cecum counts were also significantly higher in the fasted birds in one trial and liver/spleen and cecum counts in the second. No differences in SE counts were observed in unmolted vs wheat middling birds. These results indicate that there are methods alternative to feed removal which can be used to molt birds and not increase the risk for SE problems. How Commercially available microbiological systems are less expensive to use than conventional method of identification. These methods also yield relatively rapid results when compared to conventional methods. This study was conducted to evaluate the effectiveness of the relatively new BBL Crystal Enteric/Nonfermenter System (Crystal) for identification of gram negative microorganisms isolated from commercial poultry operations. The API20E (API) is a well-established method for identification of gram negative organisms from poultry. For purposes of this study, 50 isolates were obtained from commercial operations, cultured and identified by an established laboratory using the API system. The same isolates were subsequently identified using the Crystal systems in our laboratory. The Crystal system is useful for the screening of isolates from poultry sources. Key Words: API 20E, BBL Crystal E/NF, Poultry Microbiology, Diagnostic Technique 103 The effects of early handling on disease resistance of turkeys given repeated dexamethasone treatments. G.R. Huff*, W.E. Huff, J.M. Balog, and N.C. Rath, USDA/ARS/PPPSRU, Fayetteville, AR /USA. When rat pups are handled daily during the first week of life, the stress response is permanently altered resulting in increased resistance to stressinduced immunosuppression. We have developed a model for reproducing Escherichia coli air sacculitis and turkey osteomyelitis complex (TOC) using immunosuppression with dexamethasone(DEX). The purpose of this study was to determine if early handling can permanently affect the stress response and disease resistance of turkeys using this model. Eight hundred male turkey poults were handled 0, 1, or 2 times daily for the first 10 d after hatch. Starting on d 11, half of the birds from each handling treatment were treated with 3 injections of 2 mg/kg DEX on alternating days. On the day of the third DEX treatment, duplicate pens of birds were also inoculated in the air sac with 0 or 50 cfu of /italicizeE. coli. The same birds were treated with a second series of DEX injections at 5 wk of age and 4 birds/pen were bled and 10 birds/pen were euthanized 2 wk later. Surviving birds were treated with a third series of DEX injections at 10 wk of age. Two wk later,4 birds/pen were bled and all surviving turkeys were euthanized. Total and differential white blood cell (WBC) counts were determined. All mortalities and euthanized birds were scored for air sacculitis, and examined for TOC lesions. All livers, air sacs, and TOC lesions were cultured for bacteria. There was increased mortality in 2X handled birds after the first series of DEX treatments. After the second series of DEX treatments, 1X handled birds had decreased BW, and increases in mortality, air sacculitis, WBC, heterophil/lymphocyte ratio (H/L), and recovery of E. coli from tissues, whereas 2X handled birds were identical to non-handled controls. After the third series of DEX treatments handling either 1X or 2X resulted in increased BW. Handling 1X resulted in decreased mortality, air sacculitis scores, and incidence of green liver, TOC lesions, and recovery of E. coli from tissues. Birds handled 2X and treated with DEX had increased total WBC counts and H/L compared to non-handled DEXtreated birds. These results suggest that early handling affects disease resistance of male turkeys and that differences may be influenced by age and individual variability in the stress response. Key Words: Stress, Handling, Turkeys, Escherichia coli, Turkey Osteomyelitis Complex 104 Interleukin-1 contributes to the acute phase response in growing quail challenged with Salmonella typhimurium lipopolysaccharide. Kirk Klasing*, 1 University of California, Davis, CA. Partially purified preparations of chicken interleukin-1 (IL-1) have been implicated in a variety of acute phase responses to inflammation including 25 abstracts of papers decreased growth, fever, skeletal muscle catabolism, release of corticosterone, hepatic acute phase protein secretion, and increased antioxidant defenses. In mammals other cytokines (e.g. IL-6 and TNF) possess activities that overlap with IL-1. To determine the aspects of the acute phase response that can be attributed to IL-1, we used the ligand-binding domain of the chicken IL-1 type-I receptor (sIL-1R) to neutralize IL-1 activity in vivo. sIL-1R was produced in Pichia pastoris and was previously shown to block IL-1 activity as defined by thymocyte co-mitogenesis with PHA (Poultry Sci 74: 160). Japanese quail (BW=7.3 g; n=6/trt) were given ad libitum access to feed and injected i.p. with either saline (Control), 1 mg/kg S. typhimurium lipopolysaccharide (LPS), or LPS + 10 µg sIL-1R. Blood plasma was collected 12 hrs post-LPS and was analyzed for glucocorticoids (G), zinc (Zn) and the positive acute phase proteins α-1-acid glycoprotein (AGP) and transferrin (TF). Liver cytosol was analyzed for metallothionein (MT) and glutathione (GSH) as an index of antioxidant defense. Muscle cytosol was analyzed for 3-methyhistidine (3MH) as an index of catabolism. Data in the table below are expressed as % of Control and indicate that IL-1 significantly (+ = P<0.05) contributes to most aspects of the acute phase response of quail. A notable exception is that LPS did not induce a statistically significant (-) increase in GSH unless IL-1 was neutralized. Treatment G Zn AGP TF MT GSH 3MH LPS LPS+sIL-1R LPS vs C sIL-1R vs C 195 136 + + 74 92 + - 226 144 + + 149 129 + - 166 109 + - 119 133 + 137 97 + - Key Words: interleukin-1, soluble receptor, acute phase response, immunity, quail Saturday, AM, POSTERS 105 Can peripheral quantitative computed tomography be used to predict bone ash in broiler chickens?. W.D. Clark*, E.L. Smith, and M.E. Cook, University of Wisconsin, Madison, WI, USA. Peripheral quantitative computed tomography (CT) was used to evaluate the usefulness of CT in predicting whole bone ash in male broiler chickens ex vivo. The purpose of this study was to assess the feasibility of in vivo longitudinal studies using CT to examine bone mineral. Data collected in this study were a subset from a larger study testing transgenic alfalfa phytase. Six diets were formulated to contain 0.40, 0.45, 0.50, 0.55, 0.60, or 0.65 % total phosphorus. Calcium was adjusted to maintain a Ca:total P ratio of 1.4. Right tibiae were collected from 3 wk old birds, cleaned and frozen until analysis. Of 54 bones collected (9 to 11/diet except for 0.40% P where n=4), 51 were scanned using a Stratec XCT960A densitometer (Norland/Stratec). In all bones, four serial 1.25 mm cross-sectional slices were scanned in the center of the diaphysis. In a subset of 38 bones (7/diet except for 0.40% P where n=3), two additional slices were scanned on each side of the four in the center. Bone mineral content (BMC, mg/mm) and density (BMD, mg/ccm) were determined for each slice. Following CT scanning, the bones were ether extracted and ashed. Percent fat-free ash (%ASH, DM basis) of the whole bone was calculated. Various combinations of four to eight of the slices were averaged to estimate BMC and BMD for 5 to 10 mm segments of bone. Regression analyses were used to compare CT results vs %ASH. The effect of dietary P on %ASH was also analyzed. Results for 11 slice combinations in the subset (n=38) showed similar slopes and adjusted-r2 values to each other and the whole data set (n=51) when %ASH was regressed against BMD. BMD predictions of %ASH were 75 to 78% (P<.0001). A combination of BMD and dietary P increased the predictability of %ASH to 80 to 83% (P<.01). This was higher than dietary P alone (n=54, adjusted-r2 =63.9%, P<.0001). The results indicate that %ASH for the whole bone can be predicted from BMD in the diaphysis of an excised tibia, and it would be feasible to use CT in longitudinal studies in chickens. Key Words: Broiler chicken, Bone ash, Computed tomography, CT, Phosphorus 106 Effects of 25-hydroxycholecalciferol on the ability of turkey poults to utilize phosphorus from a typical corn-soybean meal diet. G. M. Owens* and D. R. Ledoux, University of Missouri, Columbia, MO. A study was conducted to determine the effects of 25hydroxycholecalciferol (25-OH-D3 ) on performance and bone mineralization in poults. Two hundred day-old male poults were randomly allotted to 8 treatments (5 reps of 5 poults each) for 21 days using a 2 x 4 factorial design. A basal diet (A) containing 0.60% available phosphorus (aP) and 1.2% Ca was prepared for diets 1, 2, 3, 4, and a basal diet (B) containing 0.45% aP and 1.2% Ca was prepared for diets 5, 6, 7, 8. The dietary treatments included: (1) A + 60 mg/ton 25-OH-D3 ; (2) A + 75 mg/ton 25-OH-D3 ; (3) A + 90 mg/ton 25-OH-D3 ; (4) A + 105 mg/ton 25-OH-D3 ; (5) B + 60 mg/ton 25-OH-D3 ; (6) B + 75 mg/ton 25-OH-D3 ; (7) B + 90 mg/ton 25-OH-D3 ; (8) B + 105 mg/ton 25-OH-D3 . With the exception of P the diets met or exceeded the nutrient requirements of turkeys (NRC, 1994). Feed intake was not affected (P > .05) by level of 25-OH-D3 but was higher (P < .05) in poults fed 0.60% aP compared with poults fed 0.45% aP. Body weight gain was affected (P < .05) by P level but not (P > .05) by level of 25-OH-D3 . Feed conversion was not affected (P > .05) by dietary treatments. Toe and tibia ash were affected (P < .05) by P level, but not (P > .05) by level of 25-OH-D3 . No P level by 25-OH-D3 level interaction was observed for any of the performance variables or bone ash. P retention was not affected (P > .05) by P level or by 25-OH-D3 level. However, a significant (P < .05) P level by 25-OH-D3 interaction was observed, indicating that the P retention response was not the same at both levels of aP. Bone breaking strength was affected (P < .05) by P level, but was not affected (P > .05) by 25-OH-D3 level nor was there a P level by 25-OH-D3 level interaction. Results indicate that levels of 25-OH-D3 ranging from 60-105 mg/ton were equally effective in supporting poult performance, bone ash, and bone breaking strength when added to diets containing 0.60% or 0.45% aP. Key Words: Poults, 25-hydroxycholecalciferol, Phosphorus, Bone ash, Performance 107 Dietary sinapic acid does not affect growth and organ weights of broiler chickens. Hongyu Qiao*1 and H. L. Classen1 , 1 University of Saskatchewan, Saskatoon, SK, Canada. Sinapic acid is a major phenolic acid in canola meal where it is primarily found as a portion of the compound, sinapine. Sinapine is composed of sinapic acid bound to choline via an ester linkage. Sinapine and sinapic acid are thought to contribute to the dark color, bitter taste, and astringency of canola meal and thereby affect meal quality. Despite the speculation that sinapic acid is an anti-nutritional factor, little research has been completed on this topic. Therefore, the objective of this research was to study the effect of dietary sinapic acid on the productivity and relative organ weights of broiler chickens. A corn-soybean diet was supplemented with 5 levels of sinapic acid (0, 0.05%, 0.10%, 0.15%, 0.20%) which are equivalent to the sinapic acid content of diets containing 0, 7.5%, 15%, 22.5% and 30% canola meal. Each treatment was replicated 4 times with 4 male birds in each replication. Production parameters were monitored from 0 to 18 days of age after which the birds were sacrificed to determine relative organ weights. Dietary level of sinapic acid was found to have no effect (P>0.05) on body weight gain, feed intake or feed efficiency ratio. Similarly, level of sinapic acid did not affect bursa of Fabricius, heart, kidney, liver, spleen, pancreas, proventriculus, gizzard, small intestine or ceca weights. In conclusion, low levels of dietary sinapic acid do not have an anti-nutritional effect in broiler chickens as indicated by performance and organ weights. Key Words: Sinapic acid, Growth, Broiler chickens, Canola 108 Citrus pulp pellet as material for broiler litter. J. O. B. Sorbara*, M. F. Rizzo, A. C. Laurentiz, and V. M. B. Moraes, Universidade Estadual Paulista, Campus de Jaboticabal - Sao Paulo Brazil. The citrus pulp pellet (CPP) is a by-product of the industry of orange juice. The search of economic alternative materials for broiler litter made possible the use of CPP. It does not have enough information about the characteristics and risks of use this by-product. The objective of this experiment was evaluate the effect of CPP as broiler litter in different stocking densities and depth of litter. It was used 1792 males Avian