Bioctive milk components to secure growth nd gut development in preterm pigs ESTER ARÉVALO SUREDA PIGUTNET FA1401 STSM
STSM Pigutnet FA1401 STSM 03/Septemer 30/Novemer/2017 (3 months) Host: Home: Thoms Thymnn Deprtment of Veterinry nd Animl Sciences Section for Comprtive Peditrics nd Nutrition University of Copenhgen, Denmrk Björn Weström Deprtment of Biology Lund University, Sweden 2
OUTLINE BACKGROUND STSM AIMS STSM TIMELINE STSM TASKS LEARNED / SKILLS ACQUIRED STSM OUTCOMES 3
Bckground PhD thesis: Development of the gstrointestinl trct in young mmmls: Effects of enterl provoction with protese or phytohemgglutinin (PHA) in neontl rts Postntl gut mturtion precociously induced y luminl stimuli Switch to n dult type intestinl epithelium with chnged digestive nd sorptive properties, incresed rrier properties nd incresed pncretic function 4
Bckground Orlly dministered pncretic nd pncretic like enzymes: Proteses provoked precocious gut mturtion Other enzymtic ctivities hd no effects Protese nd PHA induced precocious gut mturtion Mimic chnges occurring nturlly t wening Decresed intestinl mcromoleculr permeility Incresed lekiness temporrily Stimultion of pncretic function 5
STSM Aims Prcticl trining on lortory niml hndling in experiments on pre term pigs Become n ctive memer of the tem prticipting in the two ig experiments: 1. Protein fortifiction (ovine colostrum) of se diet for preterm pigs Evlution of ovine colostrum protein fortifiction of se diet for pre terms. It hs een found out tht donor s milk hs low protein content. Even though there is commercilly ville humn protein fortifier, the costs re very high. Thus, ovine origin fortifier could e used in replcement nd mke it commercilly ccessile. 2. Testing of ioctive component from the compny Arl Foods Ingredients To monitor gstrointestinl function sttus for the potentil helth enefits of the dietry fctor 6
STSM Timeline EXPERIMENT 1 5 weeks rek 2 weeks EXPERIMENT 2 5 weeks 7
STSM Tsks lerned / skills cquired During cesren sections: ring neworn piglets to NICU, monitor rething nd temperture until stilistion Dily routines: ody weight, dosge, dt recording, enterl feeding (every 2/3 hours), PN lines, dministering i.v. oluses when needed CLINICAL nd feces scoring to ssess gut nd generl helth sttus In vivo Tests Assistnce: glctose sorption test nd Behviour tests: trough drinking trining, room nd hndling hitution, ssisting during ehviour tests Smpling orgns: gut, other internl orgns, rin, etc. s well s lood collection from the jugulr vein 8
University of Copenhgen FACULTY OF HEALTH AND MEDICAL SCIENCE, Deprtment of Veterinry nd Animl Science Pncretic enzyme mturtion is delyed nd not ffected y the first enterl nutrition in preterm pigs STSM Outcome 51 st nnul meeting ESPGHAN 2018 Geneve, Switzerlnd Astrct Poster presenttion Ester Arévlo Sured 1*, Björn Weström 1, Per Torp Sngild 2, Thoms Thymnn 2 1 Lund University, Biology, Lund, Sweden 2 University of Copenhgen, Comprtive Peditrics nd Nutrition, Copenhgen, Denmrk *ester..sured@icloud.com, thoms.thymnn@sund.ku.dk Ojectives: Preterm infnts hve n immture digestive system, predisposing them to short- nd long-term complictions, including food intolernce nd necrotising enterocolitis (NEC). Pncretic enzymes re importnt for digestion ut little is known out the postntl development of the exocrine pncres, nd how it responds to enterl nutrition. Using piglet model, our ojective ws to compre the exocrine pncres of preterm nd term pigs, fed either miniml enterl nutrition (MEN) or totl prenterl nutrition (TPN) for the first 5 dys fter irth. Hypothesis: - Postntl development of the exocrine pncres is delyed fter preterm irth. - Miniml enterl nutrition during the first dys of life provides etter stimultion of the exocrine pncres function reltive to totl prenterl nutrition. Conclusions: Pncretic trypsin nd mylse reltive ctivities incresed with ge in preterm piglets, nd y dy 26 the enzyme levels converged to those of term piglets. TPN or MEN nutrition for the first 5 dys only hd trnsient effects on enzyme levels in term piglets, indicting tht the effect of enterl nutrition is ge-dependent. Incresed trypsin ctivity in term, 5 d-old TPN-fed piglets my e explined y lck of enterl stimuli to stimulte pncretic secretion. Conversely, mylse ctivity level ws very low t irth nd incresed with ge. Lower mylse ctivity in term, 5 d-old TPN-fed piglets my e explined y lck of enterl stimuli to increse enzyme production. Pncretic trypsin nd mylse ctivities depend on oth gesttionl ge t irth nd postntl ge nd hve different sensitivities to the first feeding. Methods: Pigs were delivered y cesren section preterm (90% gesttion) or t full term, nd were nurtured during the first 5 dys with totl prenterl nutrition (TPN) or with prenterl nutrition plus miniml enterl nutrition (MEN) with ovine colostrum. From dy 6, ll pigs were fed full enterl nutrition with ovine milk until postntl dy 26. Pncretic smples were collected on dys 1, 5 or 26 from oth preterm nd term pigs (n=116). Trypsin nd mylse ctivities were nlyzed in tissue homogentes. Protein content ws mesured nd used to clculte the reltive mount of the enzymes. Dt were nlyzed for the sme gesttionl ge etween tretments (e.g. MEN vs TPN, Tukey's post-hoc test, P< 0.05 indicted with different letters) nd within tretments etween different gesttionl ges (e.g. Preterm vs Term, Sidk's post-hoc test, P< 0.05 indicted with *). Trypsin (U/g protein) Am ylse (U/g protein) 100 80 60 40 20 0 400 300 200 100 0 A g e (d ): 0 dys M E N T P N M E N T P N M E N T P N M E N T P N 5 dys 26 dys ** **** 0 dys *** M E N T P N M E N T P N ** M E N T P N M E N T P N 5 dys 26 dys c c Preterm Term Results: Trypsin nd mylse ctivity incresed with ge in preterm piglets, especilly y 26 dys (P<0.05), with no differences etween the diet regimens (TPN vs MEN) (see Figure). Term piglets showed higher trypsin ctivity levels t irth nd these were lrgely mintined with dvncing ge, except for higher levels in TPNfed reltive to MEN on dy 5 (P< 0.05). Amylse ctivity ws low t irth in term piglets nd incresed with ge (P< 0.05). At 5 dys, term TPN-fed piglets hd lower mylse ctivity reltive to MEN. Trypsin ctivity ws lower in preterm thn in term piglets t 5 dys fter irth (P< 0.05) wheres mylse ws higher in preterm thn in term piglets t dy 26 (P< 0.05). The uthors declre no conflict of interest Acknowledgements: Coopertion project etween University of Copenhgen nd Lund University funded y the COST Action FA 1401 Pigutnet 9
STSM Outcomes Mnuscript in preprtion Development of exocrine pncretic function in erly life in pigs nd how it cn e influenced y diet. Pncretic tissue collected t Copenhgen, Denmrk Anlysis of pncretic tissue for peptidse, mylse nd lipse ctivity in Lund, Sweden Additionl dt to e included on expression of PLRP 2 nlysed in St. Louis, USA 10
STSM Outcomes Mnuscript in preprtion EXPERIMENT 1 Postconception vs. postntl ge (PRETERM TERM) EXPERIMENT 2 Prenterl Nutrition vs. Miniml Enterl Nutrition (TPN) (MEN) PRETERM vs. TERM EXPERIMENT 3 Bovine colostrum fortifiction vs. rw ovine milk in preterm piglets HYPOTHESIS Postntl development of the exocrine pncres is delyed fter preterm irth Miniml enterl nutrition during the first dys of life provides etter stimultion of the exocrine pncres function reltive to totl prenterl nutrition Bovine colostrum fortifiction stimultes exocrine pncres function 11
Post conception vs. Postntl ge Amylse (U/g protein) Trypsin (U/g protein) 50 40 30 20 10 0 3000 2000 1000 0 Preterm (1d) Preterm(11d) Term (1d) Term (11d) Preterm (1d) Preterm(11d) Term (1d) Term (11d) 100 Lipse (mu/g protein) 80 60 40 20 0 c Preterm (1d) Preterm (11d) Term (1d) Term (11d) 12
Prenterl Nutrition (TPN) vs. Miniml Enterl Nutrition (MEN) MEN: Miniml Enterl Nutrition TPN: Totl Prenterl Nutrition 13
Bovine colostrum fortifiction vs. rw ovine milk in preterm piglets Amylse (U/g protein) Trypsin (U/g protein) 100 80 60 40 20 0 15000 10000 5000 COLOSTRUM **** CONTROL 0 COLOSTRUM CONTROL 60 MEN: Miniml Enterl Nutrition PN: Prenterl Nutrition ENT: Enterl Nutrition Lipse (mu/g protein) 40 20 * 0 COLOSTRUM CONTROL 14
STSM Outcomes Mnuscript in preprtion CONCLUSIONS Pncretic enzymes ctivities depend on oth gesttionl ge t irth nd postntl ge Effect of enterl nutrition is ge dependent TPN or MEN nutrition for the first 5 dys only hd trnsient effects on enzyme levels in term piglets TPN lck of enterl stimuli hs different effects on different pncretic enzyme ctivities Pncretic enzyme ctivities hve different sensitivities to the first feeding 15
DIETARY MODULATION MICROBIOTA PROTEASES EXOCRINE PANCREATIC FUNCTION: DIGESTION ANTIMICROBIAL GUT HEALTH Lipse GUT DEVELOPMENT Amylse Peptidse 16
THANKS FOR YOUR ATTENTION 17