4/16/2009. Gabler Laboratory Research Program. Long chain n-3 Fatty Acids, Fetal Programming and Swine. My Background. Swine Research Areas

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1 /1/9 Gler Lortory Reserch Progrm Nichols Gler Venktesh Mni (PhD. student) Emily Kuntz (BSc. student) Mrth Jeffrey (L. Mnger) Deprtment of Animl Sciences Iow Stte University L Troe University B. Agriculturl Science Ph.D. My Bckground Purdue nd Iow Stte University Postdoctorl reserch ssocite Iow Stte University Fll 8 Animl Science fculty 3%Reserch 7% Reserch Feed efficiency Swine Reserch Ares Mternl nutrition nd fetl progrmming Gstrointestinl test trct Helth Nutrient trnsport Endotoxin trnsport Mycotoxin trnsport Skeletl muscle protein synthesis nd degrdtion Swine Residul Feed Intke (RFI) RFI: The difference etween ctul feed intke nd tht predicted on the sis of requirements for production nd mintennce of ody weight (Feed consumed over nd ove expected requirements for production nd mintennce) Yorkshire Pigs Control (rndom reed) Select (low RFI, high efficiency selection reed) AFRI-USDA 9 Growth nd Nutrient Utiliztion Grnt Aim #1: Evlute dietry nutrient retention, energy lnce, protein nd len ccretion rtes in finisher pigs selected for low RFI Aim #: Determine the extent to which ppetite regultory peptides contriute t to ltered feeding ehvior, reduce feed intke nd enhnces feed efficiency in low RFI pigs Aim #3: Identify key metolite, srcoplsmic nd mitochondril proteins tht correlte with improved skeletl muscle metolism nd ccretion in finisher pigs selected for low RFI Long chin n-3 Ftty Acids, Fetl Progrmming nd Swine 1

2 /1/9 Suckling nd Wenling Pig Chllenges Hypothesis The neworn experiences life-thretening situtions which re excerted y its physicl wekness nd low energy reserves Mternl diet influences the ftty cid profile of milk nd the piglet Birth Wening % of totl clories FAT PROTEIN CARBOHYDRATE Incresing the n-3 ftty cid content of piglet tissues vi the mternl diet will improve intestinl morphology nd glucose uptke t the wening trnsition Gler et l. (7) Journl of Nutrition 137(11): Gler et l. (9) Journl of Nutritionl Biochemistry (1): 17-5 Sow Milk Ftty Acid Profile n-3 Ftty Acid Concentrtions in Piglet Smll Intestinl Tissue CONT PFO (g/1g totl ftty cid) α-la.1.57 EPA..7 DPA..7 DHA.. EPA CONT PFO DHAGF DHA CONT PFO DHAGF Totl n Totl n PFO = Protected Fish Oil rich in DHA nd EPA 1.. CONT PFO DHAGF Different letters within rows represent significnt differences etween tretments (P<.5) Mens±SEM, n = milk smples from different sows/tretment Different letters, represent significnt differences etween tretments (P<.5) Mens±SEM, n = piglets/tretment Jejunum Morphology ws not Altered y n-3 Ftty Acids Intestinl Glucose Trnsport CONT PFO SEM P-vlue Villus height (μm) Crypt depth (μm) Villus:Crypt rtio Ussing Chmers used to mesure ctive glucose trnsport Chnge in current cross the intestine High chnge in current = Incresed Active Glucose Trnsport Mucos uffer solution seros uffer solution Mucos Seros

3 /1/9 n-3 Ftty Acids Increse Intestinl Glucose Trnsport t Wening Intestinl Glucose Trnsporters Active glucose uptke Cont/Cont PFO/PFO PFO/Cont Cont/PFO High Intestinl Glucose ENTEROCYTE Lumen N+ Glucose N+ SGLT1 GLUT Blood strem Apicl memrne Sodiumdependent Glucose Trnsporter (SGLT1) Bsolterl Glucose Trnsporter (GLUT) GLUT Different letters, represent significnt differences etween tretments (P<.5) Mens±SEM, n = 5 piglets/tretment Apicl memrne MUCOSAL Bsolterl memrne SEROSAL n-3 Ftty Acids Increse Glucose Trnsporter Protein Where to From Here? Memrne GLUT Aritrry Units Cont/ContPFO/PFO Cont/PFOPFO/Cont SGLT1 (Totl) 35 ts Aritrry Unit Cont/Cont PFO/PFO Cont/PFO PFO/Cont Wht is hppening to mino cid trnsporter? Link etween AMPK nd ATPse? AMPK or ATPse knockdown kd studies in pig enterocytes Do fll off pigs hve different nutrient trnsport cpcities thn rigorous littermtes? Different letters represent significnce (P<.5) Innte Immunity nd n-3 Ftty Acids Toll like receptor (Tlr) ntgonized y n-3 ftty cids (Gler et l., 8) n-3 ftty cids re le to lock LPS trnsmemrne signling (Chu et l., 1999) Anti-inflmmtory properties Toll like receptor signling pthwy LPS CD1 TLR MD Myd88 IRAK TRAF TRAF IKKβ IκBαNFκB TNF,IL IκBα NFκB NFkB pthwy regultes inflmmtion Required for innte immune responses NFkB IL TNFα Pro-inflmmtory cytokine production nd rective oxygen species (ROS) 3

4 /1/9 n-3 Ftty Acids Attenute the Ferile Response to LPS Questions We Are Asking? emperture (C) Rectl te Time P< Tretment P=.8 d Tretment*Time P<.1 1. d c c c.5 c c c ccc (Sline) ccc (LPS). ppp (LPS) 39.5 pcc (LPS) Role of Tlr, CD1? Mycotoxin? Dirrhe, E. coli F18 nd K88 in wened pigs? Endotoxin nd its impct on nutrient trnsport? Cn n-3 ftty cids ttenute endotoxin trnsfer cross the intestinl mucos? Hours post chllenge Pigs (3.5 ± 3. kg) were chllenged with 1 μg/kg BW of lipopolyscchride (LPS) or sline y intrmusculr injection. n = 1 pigs per tretment. Intestinl Endotoxin nd Mycotoxin Trnsport The trnsepithelil trnsport of fluorescein isothiocynte leled LPS (FITC-LPS) or Afltoxin B1 nd B The effects of ftty cids nd fier on pig intestinl endotoxin nd mycotoxin trnsport Mounted jejunum smples from pigs re mounted into Ussing chmers The mounted intestines will e incuted with FITC-LPS or mycotoxin on the mucosl side Aliquots of medium from the mucosl nd serosl sides will e tken t 15 min intervls for up to 1 min nd the luorescence of the liquots determined in plte reder The pprent permetion coefficient (Ppp) will e clculted using the following eqution: Ppp = dq/(dt x A x C ) Where dq/dt is the trnsport rte nd corresponds to the slope of the regression line, C is the initil concentrtion in the donor chmer, nd A is the surfce re of the tissue EMILY KUNTZ INDEPENDENT RESEARCH PROJECT (ANS 9) DEPARTMENT OF ANIMAL SCIENCES IOWA STATE UNIVERSITY Bckground Endotoxin The digestive trct is mjor entry point for pthogens The intestines form mjor physicl rrier to pthogens from entering circultion nd then ctivting the cquired nd innte immune systems Bcteri (endotoxin) Fungl mycotoxin Other toxic compounds Immune chllenges (cute or chronic) directly impct the growth nd reproductive performnce of swine Endotoxin cn e derived from grm negtive cteri such s E. coli nd Slmonell Endotoxin ctivtes receptor medited inflmmtory processes Endotoxin is sored y the intestine Prcellulr trnsport through the tight junctions Trnscellulr pthwy vi pssive diffusion through the enterocyte (sorptive cells) This sorption of the gut-derived cteri endotoxin plys n importnt role in the development of intestinl dysfunction, inflmmtion, peripherl tissue ctolism nd metolic disese

5 /1/9 Mycotoxin Diet, Endotoxin nd Mycotoxin Trnsport The mycotoxin is commonly found in cerels nd cn cuse: Vomiting Porcine pulmonry edem Anorexi Slow growth rtes Necrosis Decresed protein synthesis Dirrhoe Mlsorption of nutrients Reduced reproductive performnce Mortlity Compromised immunity in pigs Mycotoxin Afltoxin B1 & B Deoxynivlenol (Vomitoxin) Zerlnone Tricothecene Do different dietry compound effect the rte of trnsport of endotoxin nd mycotoxin? Dietry ftty cids Sturted ftty cids (coconut oil) Unsturted ftty cids (corn oil, olive oil, veg. oil, fish oil) Dietry fier Strch Cellulose Citrus pectin FITC-LPS Trnsport nd Ftty Acids (1) FITC-LPS Trnsport nd Ftty Acids () FITC C-LPS (rfu) Control Coconut Fish Olive Oil Cod Liver Vegetle Ppp (cm/min) c c FITC-LPS (rfu) Control CornOil Cod CLA Ppp (cm/min) c Control Corn Oil Cod Liver CLA Different letters represent significnce (P<.5) Different letters represent significnce (P<.5) FITC-LPS Trnsport nd Fier Mycotoxin FITC-LPS (rfu) Control Pectin Strch Cellulose Ppp (cm/min) FITC-LPS (rfu) F Control CornOil CodLiver CLA 1 Ppp (cm/min) 1 8 Different letters represent significnce (P<.5) Control Pectin Strch Cellulose Different letters represent significnce (P<.5) Control Corn Oil Cod Liver CLA 5

6 /1/9 Summry Conclusion Sturted fts increses endotoxin trnsport rtes n-3 ftty cids (fish oils) decrese endotoxin trnsport Conjugted linoleic cid ugments oth intestinl endotoxin nd mycotoxin trnsport Potto strch ids in the trnsport of endotoxin cross the intestine, while cellulose decreses this trnsport Dietry ft differentilly lters fltoxin trnsport Dietry compounds could e utilized to decrese the mounts of endotoxin nd mycotoxin trnsport cross the intestine Better understnding of how different dietry compounds influence endotoxin nd mycotoxin could led to etter diet formultions nd improved growth nd reproductive performnce in swine Further reserch needs to confirm this hypothesis