dr. jeffrey escobar - does immune activation alter growth potential and nutrient digestibility?
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Does Immune Activation Alter Growth Potential and Nutrient Digestibility? - Dr. Jeffrey Escobar, Novus, from the 2014 Allen D. Leman Swine Conference, September 15-16, 2014, St. Paul, Minnesota, USA. More presentations at http://www.swinecast.com/2014-leman-swine-conference-materialTRANSCRIPT
Alterations in amino acid digestion and metabolism
during diseaseJeffery Escobar, Ph.D.
Executive ManagerNutritional Physiology Research
Agenda
• Immune activation and metabolism alteration
• Plasma amino acids
– Changes during LPS challenge and their balance
• Amino acid digestibility
– Apparent, basal ENL, standardized
– Salmonella challenge
– Fate of undigested CP/AA
• Conclusions
LPS induce inflammatory response
• Inflammatory cytokines
– TNF-α, IL-1b, IL-6
• Induce fever
• Reduce feed intake
• Stimulate – Skeletal muscle proteolysis
– Liver protein synthesis
• Produce acute phase proteins
Kawai and Akira, 2010
Protein synthesis and accretion in immune challenged pigs
Orellana et al., 2002; Escobar et al., 2004
* P < 0.05
Immune activation and metabolism
mg/kg BWAA Nitrogen
Phe 0 0Trp 2 <1Val 10 5Ser 11 1Cys 12 1Tyr 16 1Pro 25 3Met 26 2Thr 28 4Gly 39 7Ile 41 4Asp+Asn 61 12Ala 66 10His 68 11Leu 72 8Arg 83 28Lys 104 20Glu+Gln 140 15
Excess release of AA and nitrogen from skeletal muscle protein over incorporation into mixed acute-phase proteins
– Increase in plasma urea nitrogen
Reeds et al., 1994
• Compare AA composition of skeletal muscle protein and a mixture of acute-phase proteins
• Assuming a typical increase in the synthesis of a mixture of acute-phase protein of 850 mg/kg BW
• The amount of skeletal muscle protein needed to be mobilized = 1,980 mg
• A difference of 1,150 mg/kg BW
• For a 100 kg pig:
– About 200 g of muscle protein mobilized
– About 13 g of excess nitrogen excreted
LPS, cytokines, and plasma urea NP
lasm
a I
L-6
, p
g/m
L
Pla
sma u
rea n
itro
gen
, m
g/d
L
Hour after-injectionHour after-injection
Webel et al., 1997
12-h feed-deprived pigs
Immune activation in fasted vs fed state
• Most acute immune activation studies with LPS have been conducted in the fasted state
– Normal food deprivation period: 8 - 24 h
• Although sick, pigs normally continue to eat, digest, and absorb nutrients
• Does acute immune activation in the fasted and fed state result in similar changes
– Digestion, absorption, metabolism, excretion
Arrival
24 h
LPS
-10 0
Surgery
Fasting Blood sampling
Feed
Recovery-2
Blood urea nitrogen in fed pigs
Price, 2011
Changes in plasma Phe with LPS
Price, 2011
Changes in plasma Phe and urea nitrogen with LPS in fed pigs
Price, 2011
Changes in plasma amino acids with LPS
Price, 2011
Price, 2011
Area underthe curve
Price, 2011
Area under the curve
Estimating plasma amino acid balance
• Calculate the area under the curve for each AA
– Saline and LPS treated pigs
• Use correction factor of 0.075 L plasma/kg BW to convert AUC to µmolekg BW-1
h-1 as a unit of AA metabolism (Talbot and Swenson, 1970)
• Apply the following equation:
AA balance = (AUCLPS- AUCSAL) 0.075 L 1kg BW h2××
Price, 2011
Negative AA balance: SAL > LPS
Price, 2011
Positive AA balance: LPS > SAL
Price, 2011
Summary of AA metabolism
Is lysine the most limiting AA during immune activation?
• Acute immune activation via LPS
– Induce similar changes in plasma urea nitrogen during the fasted and postprandial state.
– Reduces the digestion and/or absorption of dietary nutrients or increases AA catabolism
– Alters plasma balance of amino acids
• Lysine was the most negative
• Alanine was the most positive
Phe to Tyr conversion
Harper’s Biochemistry; Kilani et al., 1995; Hsu et al., 2006
Changes in plasma Tyr with LPS
Price, 2011
Changes in plasma Phe to Tyr
Price, 2011
Amino acids:nutritional considerations
Rethink AA limitation order for metabolic purposes
• Dietary amino acids:
– Essential, conditional, nonessential
• “One which cannot be synthesized by the animal organism out of materials ordinarily available to the cells at a speed commensurate with the demands for normal growth” W.C. Rose, 1946
• Essentiality of amino acid for protein synthesis
– Which one is the most limiting AA?, second?
van de Waterbeemd and Gifford, 2003
Digestibility vs. bioavailability
EndogenousFactors
Species
Breed/strain
Gender
Physiological state
ExogenousFactors
Dietary AA balance
Anti-nutritional factors
Intake
Pathogens, disease?
Factors affecting AA digestibility
Salmonella challenge in pigs
• Euthanized pigs (n=12) 24 and 72 h after oral inoculation to collect ileal digesta samples
• Salmonella Typhimurium DT104
– Nalidixic acid: 25 mg/mL
– Novobiocin: 20 mg/mL
• Intra-nasal inoculation 9.8×109 cfu
Lee, 2012
Apparent ileal AA digestibility, %24 h after Salmonella inoculation
Lee, 2012
Basal endogenous AA losses24 h after Salmonella inoculation
Lee, 2012
Standardized ileal AA digestibility, %24 h after Salmonella inoculation
Lee, 2012
Salmonella challenge in ileal cannulated pigs
• 8 growing pigs
• Ileal cannulation and recovery
• Intra-nasal inoculation 1.3×109 cfuSalmonella Typhimurium DT104 (NalRNovR)
• Collect ileal digesta every 8 h from0 to 72 h after inoculation
Lee, 2012
Dynamic changes in AA digestibility in Salmonella challenged pigs
Lee, 2012
Dynamic changes in AA digestibility in Salmonella challenged pigs
Lee, 2012
Summary on digestibility and disease
• Can poor digestion leads to enteric disease?
• Enteric disease results in:
– Lower apparent AA digestibility
– Higher endogenous losses of AA
• Higher gut demand for nutrients and energy
– Production of mucins, digestibe enzymes, enterocytes, etc.
• Provide highly digestible dietary proteins
Anti-nutritional effect of indigestible protein in the hindgut
Hindgut AA fermentation
Meijers and Evenepoel 2011; Wikipedia
Protein fermentation metabolites limiting pig growth
Yokoyama et al., 1982
Dietary protease improvesintestinal environment
Dietary protease reduces protein fermentation and prevents increase in hindgut pH.
Wang et al., 2011
Dietary protease improvesintestinal environment
Protease limits growth of undesirable bacteria and enhances growth of beneficial bacterial species.
Wang et al., 2011
General conclusions
• Immune activation alters digestion, absorption, and metabolism of amino acids
• Fermentation of undigested proteins produce undesirable bacterial metabolites and alter gut environment and microbiota
• Feed pigs highly digestible ingredients or use available technologies to maximize AA digestibility and hence minimize protein fermentation
Acknowledgements
• Students and technicians
– Dr. Kathryn Price
– Dr. Hanbae Lee
– Dr. Matthew Utt
– Elizabeth Ramirez
– Heather Totty
– Greg van Eyk
– Courtney Klotz
– Heather Reeves
– Pat Williams
• Collaborators
– Dr. Rodney Johnson
– Dr. David Baker
– Dr. William van Alstine
– Dr. Teresa Davis
– Dr. Monica Ponder
– Dr. Mark Hanigan
– Dr. Allen Harper
– Dr. Kevin Pelzer
• Virginia Tech Pratt Fellowship
Thank You!
Obrigado Merci
Gracias
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谢谢
Asante sana
Благодаря ви
Mulţumesc
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감사합니다ありがとう
Tak! Danke Schoen!