DOI: 10.5061/dryad.05qfttf53
title: Maternal and/or direct supplementation with a combination of a casein hydrolysate and yeast β-glucan on postweaning performance and intestinal health in the pig

description: A 2 × 2 factorial experiment was conducted to investigate the effect of
 maternal supplementation from day 83 of gestation and/or direct
 supplementation from weaning of a bovine casein hydrolysate plus a yeast
 β‑glucan (CH-YBG) on pig performance and intestinal health on day ten
 post‑weaning. Twenty cross bred gilts (Large White × Landrace) were
 randomly assigned to one of two dietary groups (n = 10 gilts/group): basal
 diet (basal sows) and basal diet supplemented with CH‑YBG (supplemented
 sows) from day 83 of gestation until weaning (2g/sow/day). At weaning, 120
 pigs (6 pigs/sow) were selected. The two dam groups were further divided,
 resulting in four experimental groups (10 replicates/group; 3 pigs/pen) as
 follows: 1) BB (basal sows + basal pigs); 2) BS (basal sows + supplemented
 pigs); 3) SB (supplemented sows + basal pigs); 4) SS
 (supplemented sows + supplemented pigs). Supplemented pigs were offered
 0.5g CH‑YBG/kg of feed for 10 days post-weaning. On day 10 post‑weaning, 1
 pig/pen was humanely sacrificed and samples were taken from the
 gastrointestinal tract for analysis. Pigs weaned from supplemented sows
 (SS, SB) had reduced faecal scores and incidence of diarrhoea
 (P&lt;0.05) compared to pigs weaned from basal sows (BB, BS), with SS
 pigs not displaying the transient rise in faecal scores seen in the other
 three groups from day 3 to day 10 post‑weaning (P&lt;0.05). Pigs
 weaned from supplemented sows had reduced feed intake (P&lt;0.05),
 improved feed efficiency (P&lt;0.05), increased butyrate
 concentrations (P&lt;0.05), increased abundance of Lactobacillus
 (P&lt;0.05) and decreased abundance of Enterobacteriaceae and
 Campylobacteraceae (P&lt;0.05) compared to pigs weaned from basal
 sows. In conclusion, maternal supplementation increased the abundance of
 Lactobacillus and decreased the abundance of Enterobacteriaceae and
 Campylobacteraceae while also increasing butyrate concentrations. The
 combination of maternal and direct supplementation led to pigs having the
 lowest faecal scores compared to all other groups.
description: A total of 20 pregnant gilts (Large White × Landrace genetic
 lines) were blocked according to live weight and backfat and randomly
 assigned to one of two dietary groups (<em>n</em> = 10
 gilts/group): D1) basal gestation/lactation diet (basal sows) and D2)
 basal gestation/lactation diet plus CH‑YBG (supplemented sows) from day
 83 of gestation until weaning (day 28). The CH‑YBG supplement (2.0
 g/sow/d) contained 1.0g CH and 1.0g yeast β‑glucan. The ingredient
 composition of the lactation and gestation diets are presented in Table 1.
 The gestation diet contained 140 g/kg crude protein, 13.5 MJ/kg of
 digestible energy and 4.4 g/kg of standardized ileal digestible lysine.
 The lactation diet contained 190 g/kg of crude protein, 14.5 MJ/kg of
 digestible energy and 8.5 g/kg of standardized ileal digestible lysine.
 The amino acid requirements were met relative to lysine.
 From day 83 of gestation to day 110, gilts were housed in groups
 of 10 according to their allocated treatment and were allowed into
 individual feeding crates at feeding times. On day 110, gilts were moved
 to individual farrowing pens (2.2m × 2.4m) with crates, slatted floors and
 heat pads for the piglets.  The gestation house and farrowing room
 temperature was maintained at 20°C throughout the experiment. The
 experimental supplement (CH‑YBG) was top-dressed on the gestation diet and
 added to the trough prior to feeding the lactation diet each morning (9am)
 to ensure consumption. The dams received specific amounts of feed in the
 following quantities: 2.5 kg/d of gestation diet from day 83 until day 110
 of gestation. They were fed 2.0 kg/d of lactation diet from day 111 of
 gestation until the day of farrowing and then the feed supply increased by
 1.0 kg/d until day 3 post‑farrowing and by 0.5 kg/d until day 6
 post‑farrowing. Afterwards, the sows were allowed semi-ad libitum of the
 diet, which was adjusted for each sow depending on daily intake. The sows
 were fed in 2 equal meals provided at 9am and 3pm. The sows had ad libitum
 access to drinking water throughout the experimental period. 
 On the expected farrowing date, fresh sow faecal samples, which
 did not come in contact with the floor, (approximately 10 g, (SD=1.0))
 were collected from each sow into sterile containers and stored at -20°C
 for microbial analysis. Farrowing was not induced but all were supervised
 to ensure that there was no difficulty while farrowing. At parturition,
 each piglet was individually weighed, and the number of live born piglets
 were recorded. Six piglets of average birth weight (3 boars and 3 gilts)
 were selected from each litter and ear-tagged shortly after birth. Litter
 size was adjusted shortly after birth by cross-fostering piglets within
 dietary groups to ensure that sows nursed a similar number of piglets (12
 per sow), and this was maintained throughout the suckling period. No creep
 feed was offered to the piglets throughout the lactation period, and
 piglets had no access to the sow’s feed. Piglets received an intramuscular
 injection of Fe-dextran (Ferdex 100; Medion Farma Jaja, Indonesia) on day
 7 after birth. At weaning, the 120 previously selected
 pigs (6 pigs/sow) remained in the same group defined by their dams and
 subdivided into two groups of 3 pigs, resulting in four experimental
 groups. The two factors, lactation diet and post-weaning diet, were
 arranged in a 2 × 2 factorial to provide the four experimental groups that
 were randomly assigned to replicate pens (3 pigs/pen) as follows: T1) - BB
 (basal sows + basal pigs); T2) - BS (basal sows + supplemented pigs); T3)
 - SB (supplemented sows + basal pigs); T4) - SS (supplemented sows +
 supplemented pigs). The current experimental design is presented in Fig 1.
  Pigs from the BS and SS groups were offered the CH‑YBG supplement post-weaning at a
 rate of 0.5 g/kg of feed. The starter pig performance
 study measured performance between day 0 and 10 post-weaning, where pigs
 were supplemented with or without the supplement depending on the
 experimental group, as described previously. The pigs were housed in
 groups of 3 (from original sow litter) on fully slatted pens (1.7m ×
 1.2m). The post‑weaning diet contained 210.6 g/kg crude protein, 15.0
 MJ/kg digestible energy and 12.5 g/kg standardized ileal digestible
 lysine. All amino acid requirements were met relative to lysine. The diets
 used were formulated to create the greatest nutritional challenge possible
 to the post-weaned pig in the absence of in-feed medication. These diets
 contained a high crude protein level, high level of soya bean meal and low
 level of lactose. All diets were milled on site and fed in meal form for
 10 days’ post‑weaning. No medication, zinc oxide or other growth-promoting
 agents were included in the starter diet. The ingredient composition of
 the post‑weaning diet is presented in Table 1. Feed and water were
 available ad libitum throughout the experimental period. The ambient
 environmental temperature within the houses was thermostatically
 controlled. The temperature was maintained at 30°C for the first week and
 was reduced by 2°C the following week. The pigs were weighed individually
 on the day of weaning and day 10 post-weaning. Feed intake was recorded
 per pen to calculate average daily feed intake. On day
 10 post‑weaning, one pig per pen (selected at birth) was sacrificed
 following a lethal injection of pentobarbitone sodium (Euthatal Soluion,
 200mg/ml; Merial Animal Health, Essex, UK) at a rate of 0.71 ml/kg body
 weight to the cranial vena cava to humanely euthanise the animals.
 Euthanasia was completed by a competent person in a separate room away
 from sight and sound of the other pigs. Following this, the entire
 digestive tract was surgically removed. Sections from the duodenum (10cm
 from the stomach), the jejunum (60cm from the stomach) and the ileum (15cm
 from the caecum) were excised and fixed in 10% neutral-buffered formalin.
 tissue samples were taken from the jejunum, ileum and colon to measure the
 gene expression of cytokines, digestive enzymes, nutrient transporters,
 mucins, tight junction and appetite regulators using QPCR. Tissue sections
 of 1 cm² from the duodenum, jejunum, ileum and colon were cut out, emptied
 by dissecting them along the mesentery and rinsed using sterile phosphate
 buffer saline (PBS) (Oxoid, Hampshire, UK). The tissue sections were
 stripped of the overlying smooth tissue before storage in 5ml RNAlater®
 solution (Applied Biosystems, Foster City, CA, USA) overnight at 4°C. The
 RNAlater® was removed before storing the samples at -80°C. Digesta samples
 (approximately 10g) from the caecum and colon were aseptically collected
 into sterile containers (Sarstedt, Wexford, Ireland) and immediately
 frozen for subsequent 16s rRNA sequencing and VFA analysis.