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