1. Interaction between host and nutrition – factors of importance for post weaning diarrhea
Thessaloniki · September 2018

2. Working group 2: Title: “Genetic and environmental factors to understand dysbiosis including their interaction (epigenetics)”. Today’s topic: The main drivers that influence the crosstalk between pig and gut microbiome

3. Challenges in pig production
Hyperprolific sows:
increased number of piglets per litter; heterogeneous litters, and high piglet mortality
Weaning is a stressful period for the piglet:
- adaptation to new feed and environment
- separation from the dam
increased risk of diarrhea post weaning
high consumption of antimicrobials
supra-nutritional levels of zinc oxide (ZnO)
Request for alternatives to antibiotics and ZnO

4. Weaned piglets: Treatment of gastric-intestinal sources
(pleuromutilins, tetracyklins)
Sources:
DANMAP, 2016
Udvalget for Fødevarer, Landbrug og Fiskeri, sept. 2016

5. Consumption of zinc oxide in Denmark
The European Commission has confirmed the EU-wide ban on the use of zinc oxide at medicinal levels in piglet feed, giving member states up to five years (2022) to phase it out.
Increase in veterinary prescription of ZnO (0-2 weeks post weaning).
Critical because high ZnO enhances antibiotic resistant bacteria (e.g. E. coli in gut of pigs (source: PLOS one, 2018))

6. Genetics is important for PWD development
Modified from Roubos-van den Hil (2010)
Modified from Fairbrother et al., (2005) and Heo et al. (2013)

7. Dysfunction of the gut Can we measure changes in vivo?
Disrupted gut villi and integrity:
Impaired absorption
of nutrients
Lack of epithelial barrier
function
Can we measure changes in vivo?

8. Fut-1 gene in pigs
A single guanine-to adenine mutation at nucleotide 307 in α-(1,2)-fucosyltransferase (FUT-1) gene is determinant for the susceptibility of piglets to E. coli F-18 diarrhea.
What is the influence of the host genetics on the development of the gut microbiota, and the host responses (immune system and metabolome) under non-challenged conditions?

9. Animal Model X E. Coli Sensitive E. Coli Resistant Sensitive Resistent
♀ M307G/A
♂M307A/A X
Sensitive
Resistent

10. Experimental SETUP Piasma metabolomic profile
PIOC and immune responses

12. Sugiharto et al., 2014, CanJAnim. Sci.
Infection with E. coli
Sugiharto et al., 2014, CanJAnim. Sci.

13. Sugiharto et al., 2014, CanJAnim. Sci.
Immune function
Blood, 4 days post challenge
Challenge
Diet
Challenge
Small intestine, 7 days post challenge
Sugiharto et al., 2014, CanJAnim. Sci.

14. Separation of the data between E. coli challenge (not diet)
Metabolomics profile of the plasma
PCA before challenge
PCA after challenge
Separation of the data between E. coli challenge (not diet)
Sugiharto et al., 2014, CanJAnim. Sci.

15. 1) Sugiharto et al., 2014, CanJAnim. Sci.
Metabolites identified in the metabolomics analysis (discriminating between challenged and unchallenged piglets)1
Metabolomics profile and lymphocyte subsets in the blood provided an additional insight into ETEC F4-induced inflammatory responses1.
What is the importance of microbial metabolites and lipid derived metabolites for the host immunity?
1) Sugiharto et al., 2014, CanJAnim. Sci.

16. ARA in cell membrane phospholipids
Dietary fatty acids,
(C18:2 & C18:3)
PLA2
Control by antioxidants
Or by hydrolases from commensal flora1
Release of ARA
Reactive oxygen species (ROS)
COX
LOX
Prostaglandins
(PGE2),
Prostacyclins,
Thromboxane
Upregulation of COX2 in UC/source: Lauridsen
Leukotrienes
Pathogen destruction (via phagocytosis) and respiratory burst. Processes trigger immune responses
ROS
Inflammatory response, regulation of immune response (e.g. Treg), mediate of intestinal permeability.
1) Dorrestein et al Imm. Perspective. 40

17. Effect of fat on fatty acid concentration in gut mucosa in 56 d old pigs (Lauridsen, 2010)

18. Fatty acid composition influence immune
cell responses after E. Coli-LPS stimulation
Animal fat, sunflower oil or fish oil (5% for weaners)
Møller & Lauridsen, Cytokine, 2006

19. Knowledge syntheses
‘A critical review on alternatives to antibiotics and pharmacological zinc for prevention of diarrhoea in pigs post-weaning’ (Lauridsen, C., Højberg, O., Kongsted, H., and Canibe, N, 2017) .
Aim: to investigate the possibilities for prevention of weaning diarrhea in pigs in organic and conventional production based on national and international literature.
PWD as response parameter – not proxy parameters

20. Commercial conditions:
Natural or semi-natural conditions:
12–18 wk of age (organic: 7-10 weeks).
A gradual reduction in the amount of contact
between the sow and her piglets.
A concomitant reduction in nursing frequency and milk production.
A gradual change of diet from milk to solid or semisolid feed.
Commercial conditions:
3-4 weeks.
An abrupt separation of piglets from the dam.
A sudden change in diet.
A move to a new environment.
Mixing with unfamiliar animals at a much younger age than would occur in natural conditions.

21. Conclusion of literature review
Suggested strategies to reduce PWD:
Restriction of dietary protein
Valid strategy in herds with high diarrhea incidence
Increasing weaning age
- Good possibility in organic pig production
Optimizing the hygienic status of weaning facilities
-Less documented regarding organic pig production
These strategies were included in a cost-benefit analyses.
To pinpoint their

22. Conclusion of literature review
Besides, some dietary factors gained our interest:
Organic acids, antibodies, fermented liquid feed,…., but these alternatives requires more development and research to pinpoint their efficacy against PWD.
Important to design studies with PWD as primary response variable, and to design the study to test PWD.

23. How to progress? -Intervention during suckling
- Avoid abrupt weaning/change
to solid feed
How to progress?
Moeser et al, 2018, Animal Nutrition,

24. BORN2GUT (project )
Importance of colostrum intake and birth weight and additional milk/feed during suckling for weight at weaning, enteric robustness and health of the weaned piglet (7-30 kg)
Four treatments (including dietary intervention):
Weaning at 3,5 weeks & no additional milk/feed
Weaning at 3,5 weeks & additional milk/feed
Weaning at 5 weeks & no additional milk/feed
Weaning at 5 weeks & additional milk/feed

25. Measures Individual weight of all piglets at various timepoints
Individual blood samples at various timepoints
Video of behavior (who is eating what?)
Round 2:
Colostrum intake per pig (during first 24 hrs)
Killing of piglets to obtain various samples to measure gut health
Outcome: Linking individual and early experiences to enteric robustness, immunity, behavior and health post weaning. I