XII - 12th International Symposium - Digestive Physiology of Pigs

Digestive
Physiology
of Pigs
R. Minor,* T. Dortch, L. Kloc, L. Williams, and A. Woldeghebriel,
North Carolina A&T State Universtiy, Greensboro,
North Carolina, USA.
The stress of early weaning, coupled with an immature
immune system, contributes to the development of postweaning
diarrhea a condition that is associated with an
imbalance of gut microbiota and intestinal inflammation.
Intestinal microbiota is a key contributor to the maturation
of the immune system. Studies have shown that inclusion
of probiotics and prebiotics in the maternal diet during
gestation and lactation can promote health of their offspring.
A study was conducted to specifically evaluate the impact
of oat supplementation during gestation and lactation on
the intestinal health of piglets. Sows were fed gestation
and lactation diet (containing corn and soybean meal) with
or without oat at an inclusion rate of 15%. Data relating to
piglet weight at birth and weaning, intestinal IgA content
and fecal levels of beneficial bacteria, bifidobacteria
and lactobacilli, and the pathogenic bacteria commonly
associated with post-weaning diarrhea, Escherichia coli
and Salmonella, were collected. While we report that
piglets of dams on control diet weighed significantly more
at birth (P < 0.0001), piglets born of dams who consumed
oat weighed significantly (P < 0.05) more on the day of
weaning and 28 d post weaning. Microbiological culture
analysis of fecal material collected at the time of weaning
reveled no significant differences in the level of pathogenic
and beneficial bacteria however, by d 28 post-weaning,
there was significantly (P < 0.05) more beneficial bacteria
(bifidobacteria and lactobacilli) than pathogenic bacteria
(Escherichia coli and Salmonella) in the feces of piglets
born of dams that consumed oat. Moreover, we report that
dietary inclusion of oat at 15% during lactation and gestation
resulted in increased levels of IgA in the small intestine of
piglets whose dams consumed a diet supplemented with
oat. Together these data suggest that inclusion of oat in the
gestation and lactation feed of sows and positively affect
the intestinal health of their offsprings.
Key words: prebiotic, post-weaning diarrhea,
immunoglobulins
1030 Assessment of RNA integrity in the post-mortem
pig colonic tissue ex-vivo. B. Bahar* 1,2 , J. O’Doherty 1 ,
and T. Sweeney 2 , 1 School of Agriculture and Food Science,
University College Dublin, Belfield, Dublin, Ireland, 2 School
of Veterinary Medicine, University College Dublin, Belfield,
Dublin, Ireland.
Surgical removal of porcine intestine post-mortem followed
by an ex vivo challenge is an alternative technique of
testing the anti-inflammatory effect of bioactive compounds
in the intestine of live pigs. This technique is based on the
assumption that tissue remains physiologically viable and
that the cellular RNA remains intact during the ex-vivo
period. The present experiment investigated the effect of
ex-vivo incubation of porcine colonic tissue on the quantity
and quality of total RNA over a 12-h time period. A portion
of colonic tissues of pigs (n = 6) was surgically removed
immediately post- slaughter and the overlying muscle layer
removed. A section of approximately 1.5 × 1.5 cm of the
XII INTERNATIONAL SYMPOSIUM ON
DIGESTIVE PHYSIOLOGY OF PIGS
49
Session I
colon was transferred into 1 mL DMEM and incubated
for 0, 3, 6 and 12 h in a humidified cell culture incubator.
Tissue samples were collected in RNAlater and processed
for RNA extraction. The quantity and quality of total RNA
were assessed on a NanoDrop Spectrophotometer and an
Agilent 2100 Bioanalyzer, respectively. Ex-vivo incubation
time had a significant effect on the quantity (P < 0.001) and
quality (P < 0.001) of total RNA in colonic tissue. Compared
with RNA yield at the 0 h time point (505.0 ± 84.25μg/
mg of tissue), the yield was numerically reduced after 3 h
(367.2 ± 23.13 μg/mg) while RNA yield was significantly
reduced after 6h (227.6 ± 51.04μg/mg, P < 0.001) and 12
h (159.3 ± 41.91μg/mg, P < 0.001) of incubation. In the
Bioanalyzer profile of total RNA, the 28S and 18S ribosomal
RNA (rRNA) bands were visibly intact at 0, 3 and 6 h of
incubation. However, after 12 h of incubation, these 2 rRNA
populations disintegrated and a degraded RNA profile
was evident. The RNA integrity (RIN) values for the 0, 3,
6 and 12 h of incubation were 9.4 ± 0.17, 9.0 ± 0.17, 6.7 ±
0.33 (P < 0.001) and 3.3 ± 0.42 (P < 0.001), respectively
which further indicates that there is only a small reduction
in RIN values after 3 h of incubation. It is concluded that
the incubation of porcine colonic tissue ex-vivo for up to 3
h may generate good quality total RNA suitable for gene
expression studies.
Key words: RNA stability, colon, RNA integrity
1031 extracts of brown seaweeds can attenuate
the bacterial lipopolysaccharide (LPS) induced proinflammatory
response in the porcine colon ex vivo.
B. Bahar* 1,3 , J. O’Doherty 1 , M. Hayes 2 , and T. Sweeney 3 ,
1 School of Agriculture and Food Science, University College
Dublin, Belfield, Dublin, Ireland, 2 Ashtown Food Research
Centre, Teagasc, Ashtown, Dublin 15, Ireland, 3 School of
Veterinary Medicine, University College Dublin, Belfield,
Dublin 4, Ireland.
Brown seaweeds, a rich source of bioactive compounds, are
demonstrated to have numerous health benefits including
anti-microbial and immunomodulatory bioactivities in the
pig intestine. This experiment was conducted to evaluate
the effects of extracts of brown seaweed (Ascophyllum
and Fucus) in suppressing the pro-inflammatory response
induced by bacterial lipopolysaccharide (LPS) treatment
in the pig colonic tissue ex-vivo. A section of colonic
tissues of pig (n = 6) was surgically removed immediately
post-slaughter and the overlying muscle layer removed.
Approximately 1.5 × 1.5 cm of the colon was transferred into 1
mL Dulbecco’s Modified Eagle Medium containing bacterial
LPS (10 μg) and seaweed extracts (1 mg). Extracts of
Ascophyllum nodosum and Fucus serratus seaweeds were
tested in this experiment. Tissue samples were incubated
for 3 h in a humidified cell culture incubator with 5% CO 2
at 37°C. Tissue samples were collected in RNAlater and
processed for RNA extraction. Gene expression analysis
of interleukin-8 (IL8), interleukin-6 (IL6) and tumor necrosis
factor-α (TNFα) genes were performed by quantitative
real time PCR. Compared with the basal level of gene
expression, LPS treatment induced the expression of IL8,
IL6 and TNFα genes to 2.38 ± 0.86 (P < 0.01), 1.90 ± 0.66
(P < 0.05) and 1.90 ± 0.57 (P < 0.001) fold, respectively.