XII - 12th International Symposium - Digestive Physiology of Pigs

Digestive
Physiology
of Pigs
3025 Gene expression profiles of peripheral blood
mononuclear cell (PBMC) from young pigs fed high
level of zinc oxide. S. Y. Ji* 1,2 , B. H. Choi 1 , B. G. Kim 2 ,
and C. H. Yun 3 , 1 National Institute of Animal Science, RDA,
Suwon, Republic of Korea, 2 Department of Animal Science
and Environment, Konkuk University, Seoul, Republic of
Korea, 3 Protein Engineering & Comparative Immunology
Laboratory, Seoul National University, Seoul, Republic of
Korea.
This study was performed to investigate effects of high level
of zinc oxide in diets on in vivo gene expressions of PBMC
and growth performance in young pigs. Female Duroc pigs
(n = 9, 36.4 ± 4.68 kg) were assigned into 2 dietary groups.
One group (CON, n = 4) was fed the basal diet supplemented
with no ZnO, and the other group (ZN2.5K, n = 5) was fed the
basal diet supplemented with 2,500 mg ZnO/kg diet under
ad libitum feeding condition for 14 d. In the post absorptive
state on the 14th day, 8mL of blood were drawn from
jugular vein into by BD-CPT tube with sodium citrate, and
centrifuged (1,800 x g, 15 min) to obtain PBMCs from blood
samples. All of these preparations were finished within 2 h,
and then stored at −80°C until further analysis. After TRIzol
extraction of total RNAs, their A260/280, A260/A230 ratio
and RIN values were checked to be suitable for microarray
analysis. Gene expression profiles were analyzed by using
Affymetrix porcine expression array. Data were analyzed
by using GeneSpring GX11.5.1 software (1.5 fold change,
P < 0.05). Average daily gain (ADG, kg/d) was measured
for 2 weeks and did not differ (P > 0.10) between ZN2.5K
and CON (0.54 vs. 0.47 kg/d, respectively). Fifteen of
differentially expressed genes (DEG) were significantly up
(+)- or down (−)-regulated (P < 0.05, 1.5 fold change). Due
to the lack of gene descriptions from pigs, only 3 genes of 9
(+)-DEGs were described as adrenergic β receptor kinase 2
(GenBank Accession # CK459913, CF175365), and tissue
inhibitor of metalloproteinase-2 (#CK461818), which were
significantly upregulated in ZN2.5K than CON (P < 0.05).
Three of 6 (−)-DEGs were described as dynein cytoplasmic
intermediate chain 2 (#CK451466), Ca binding protein β
(neural), Sus scrofa CD3g mRNA for TcR CD3 gammachain
(#AB190229) which were downregulated in ZN2.5K
than CON (P < 0.05). In conclusion, pharmacological levels
of dietary ZnO could affect in vivo immune responses by
modulating gene expressions of PBMC in pigs.
Key words: zinc oxide, Affymetrix, PBMC
3026 Comparison of three intestinal permeability
methods. Y. van der Meer 1 , W. J. J. Gerrits* 1 , M. van den
Bosch 2 , M. Moretó 3 , W. A. Buurman 4 , and T. A. T. G. van
Kempen 5 , 1 Animal Nutrition Group, Wageningen University,
Wageningen, The Netherlands, 2 Provimi Holding B.V.,
Velddriel, The Netherlands, 3 Faculty of Pharmacy, University
of Barcelona, Barcelona, Spain, 4 Department of Surgery,
Maastricht University Medical Centre, Maastricht,
The Netherlands, 5 North Carolina State University, Raleigh,
North Carolina, USA.
The determination of intestinal permeability remains a
challenge. The Ussing chamber is an ex vivo technique
which is generally well accepted. However, this technique
XII INTERNATIONAL SYMPOSIUM ON
DIGESTIVE PHYSIOLOGY OF PIGS
128
Session VI
is tedious and requires a biopsy which can only be obtained
through invasive surgery or post-mortem. It also represents
a spot sample, sometimes less than 1 cm 2 . The everted
gut sac technique uses a more relevant sample size,
and in vivo permeability measurements using Co-EDTA
or lactulose are much more practical. Our objective was
to compare these 3 techniques. For this, 72 piglets were
sampled either 3.5, 7.5, or 10.5 d post weaning. Both
lactulose (1.3 g, 9.5 Å) and Co-EDTA (0.6 g, 10.0 Å)
dissolved in 15 g demineralized water were administered
intragastrically. Two hours later a blood sample was
obtained through venipuncture. Subsequently, a 15 cm
segment harvested at 25, 50, and 75% of the length of the
small intestine was inverted, filled with phosphate buffered
saline (PBS) containing 5 mM glucose, and sealed. Sacs
were inserted for 60 min. in an aluminum foil covered bath
with aerated PBS containing 125 µM FITC-dextran (14.0 Å)
and maintained at 39°C after which both the sac content
and medium were analyzed for FITC. Statistical analysis
was performed using the Pearson correlation test of SPSS
19 (2010). The FITC analysis of the buffer revealed that the
FITC concentrations dropped quickly (half-life ± 3 h), even
though it was kept from contact with light by aluminum foil.
This is raising doubts about the validity of the test. Serum
concentrations of Co-EDTA and lactulose were positively
correlated (r = 0.67, P < 0.01). FITC dextran, corrected for
a predicted change in half-life of stock solution, in contrast,
did not correlate with either lactulose or Co-EDTA, for any
of the segments compared. In conclusion, a correlation
was found between Co-EDTA and lactulose suggesting
that both marker molecules measure the same change
in permeability, while ex vivo determinations using FITCdextran
in everted gut sacs requires additional work before
it can be applied.
Key words: intestinal permeability, piglet, everted gut sac
3027 Flavor preferences conditioned by post-ingestive
effect of sucrose and porcine digestive peptides
(PDP) in post-weaning pigs. J. Figueroa 1 , D. Solà-Oriol* 1 ,
E. Borda 2 , S. A. Guzmán-Pino 1 , and J. F. Pérez 1 , 1 Universitat
Autònoma de Barcelona, Bellaterra, Barcelona,
Spain, 2 Bioibérica, Barcelona, Spain.
Pigs can learn to prefer a flavor if it has been previously
associated to positive consequences. The aim of this
experiment was study flavors preferences conditioned by
the post-ingestive effect of nutrients in post-weaning pigs. A
total of 240 weanling piglets were allocated in 24 pens (10
piglets/pen) and distributed to 2 experimental treatments.
G1 pigs were trained during 8 d with one flavor (CS+)
into a protein solution (4% Porcine Digestible Peptides,
PDP) in odd days and another flavor (CS-) into 100mM
of Monosodium Glutamate (MSG) solution on even days
(5 lts-bottle for 24hr). In G2, CS+ was mixed into a 4%
sucrose solution in odd days and CS- into 1% sucrose +
0.08% saccharine on even days. Hedonic attraction for
PDP and MSG solutions and attraction for sucrose and
saccharine + sucrose were previously matched, thus, in
each group one flavor (CS+) was associated with a higher
post-ingestive but the same hedonic value than the other
flavor (CS-). Solution’s amount offered during training