presentations - wur...protein for life conference ede, 25 th october 2016 lonneke...
TRANSCRIPT
Presentations
Day 2 Session 5,6
DAY 2 Session 4;5;6
Session 4 Topic: Protein and human/animal health
Annemarie Rietman Associations between dietary factors and markers of NAFLD in a general Dutch adult population
Marco Mensink True ileal amino acid digestibility of a lamb meat hydrolysate and its postprandial metabolic utilization in elderly subjects
Soumya Kar Use of multi omics approaches in the search for alternative dietary protein sources”
Meike Bouwhuis The influence of low protein diets on growth performance and carcass characteristics of gilts or entire boars fed ad libitum
Session 5 1. Learnings from human and animal studies
Lonneke Janssen Duijghuijsen Mitochondrial ATP production and intestinal epithelial permeability – an in vitro model
Hsuan Chen Protein digestion kinetics of different protein sources in pigs
Janne Beelen Effects of a 12-week intervention with protein-enriched foods and drinks on protein intake and physical performance of older patients during the first 6 months after hospital release
Lotte van der Zanden Novel protein foods: what do elderly need, what do elderly want?
Session 6 2. Side-streams and insects
Marcel Hulst In vivo and in vitro study on the mode of action of spray dried plasma when used as feed additive
Natasja Gianotten Sustainable protein production using lesser mealworms
DAY 2, SESSION 5 Session 5 Topic: 1. Learnings from human and animal studies Presentations available: • Lonneke Janssen Duijghuijsen • Hsuan Chen • Janne Beelen • Lotte van der Zanden
3
Lonneke Janssen Duijghuijsen • Mitochondrial ATP production and intestinal epithelial permeability –
an in vitro model
Novel proteins and intestinal permeability: An in vitro model
Protein for Life Conference
25th October 2016, Ede
Lonneke Janssen Duijghuijsen
IPOP Customized Nutrition
Background
6
Novel proteins
Digestion and
absorption Intestinal
permeability Screening method
Background
7
Novel proteins
Digestion and
absorption Intestinal
permeability Screening method
Transcellular Paracellular
Background
8
Novel proteins
Digestion and
absorption Intestinal
permeability Underlying mechanism
Transcellular Paracellular
Background
9
Novel proteins
Digestion and
absorption Intestinal
permeability Underlying mechanism
In vitro model Caco-2 cells
Digestion and
absorption Intestinal
permeability Screening methods
Underlying mechanism intestinal permeability
10
In vitro model Caco-2 cells
Digestion and
absorption Intestinal
permeability Screening methods
Role of mitochondrial energy production
11
High glucose in cell culture media
Role of mitochondrial energy production
Underlying mechanism intestinal permeability
12
High glucose in cell culture media
‘Crabtree effect’ (Crabtree 1929)
Role of mitochondrial energy production
Underlying mechanism intestinal permeability
13
High glucose in cell culture media
‘Crabtree effect’ (Crabtree 1929)
Energy production by glycolysis
Role of mitochondrial energy production
Underlying mechanism intestinal permeability
14
High glucose in cell culture media
‘Crabtree effect’ (Crabtree 1929)
Energy production by glycolysis
Substrate Glucose
Galactose
Role of mitochondrial energy production
Underlying mechanism intestinal permeability
(Rossignol 2004; Marroquin 2007; Aguer
2011; Kase 2013)
15
Respirometry (OROBOROS and Seahorse)
More ‘aerobic’ = more O2 consumption
Glucose versus galactose
“Aerobic switch”
16
Respirometry (OROBOROS and Seahorse)
More ‘aerobic’ = more O2 consumption
Glucose versus galactose
“Aerobic switch”
17
Western blot
More ‘aerobic’ = more mitochondrial OXPHOS complexes
Glucose versus galactose
“Aerobic switch”
18
Western blot
More ‘aerobic’ = more mitochondrial OXPHOS complexes
Glucose versus galactose
“Aerobic switch”
19
Mitochondrial staining
More ‘aerobic’ = more (green) mitochondria
Glucose versus galactose
“Aerobic switch”
20
Complex I inhibitor (Piericidin A; PA)
Cellular energy status
Mitochondrial dysfunction
Mitochondrial energy production
21
Complex I inhibitor (Piericidin A; PA)
Cellular energy status
Mitochondrial energy production
Mitochondrial dysfunction
22
Complex I inhibitor (Piericidin A; PA)
Decreased cellular energy status
Intestinal permeability
TEER and FS flux Markers for permeability
Mitochondrial dysfunction
FS
23
Complex I inhibitor
Intestinal permeability
TEER and FS flux Permeability increased
Mitochondrial dysfunction
Decreased cellular energy status
FS
Caco-2 intestinal model
Take home message (1)
Glucose Galactose
O2 O2
O2
O2
O2
O2 O2 O2
O2 O2
O2 O2
O2
O2
O2
O2 O2
O2 O2
“In vivo-like” Caco-2 intestinal model 24
25
“In vivo-like” Caco-2 intestinal model
Take home message (2)
Functional mitochondria
Dysfunctional mitochondria
FS
FS FS
FS FS
FS
Novel proteins and intestinal permeability An in vitro model
Wageningen University and Research
Jaap Keijer (HAP)
Sander Grefte (HAP)
Dorien van Dartel (HAP)
Harry Wichers (FBR)
Jurriaan Mes (FBR)
Klaske van Norren (HNE)
Renger Witkamp (HNE)
26
Protein for Life Conference Ede, 25th October 2016 Lonneke JanssenDuijghuijsen ([email protected])
IPOP Customized Nutrition
Hsuan Chen • Protein digestion kinetics of different protein sources in pigs
FAST OR SLOW Protein digestion kinetics of protein sources in pigs
Hsuan Chen, 2016-10-25
29
Current feed evaluation
The kinetics of protein digestion is not taken into account
• Current evaluation
• Composition of indispensable amino acids
• Digestibility at ileal level
Global shortage of protein sources Increased price of protein sources Increase utilization efficiency of protein sources
What is protein digestion kinetics?
30
Digestion = Hydrolysis + Transition + Secretion + Absorption + Fermentation
Digestion Kinetics
Hydrolysis rate Passage rate
Absorption rate and location
Post-absorption metabolism
31
Lumen
Blood
• Oxidation to produce energy • Protein synthesis for secretion
• Protein synthesis for muscle • Oxidation to produce energy
Free AA Hydrolysis
Protein-bound AA
absorption absorption
Amino acids absorption and metabolism
Energy source
Hydrolysis
absorption
32
Aim of the study
To determine the protein digestion kinetics of protein sources
differing in ileal digestibility in pigs
Digestion Kinetics
Post-absorption metabolism
Increase utilization efficiency of protein sources
33
Experimental setup- Design
Soybean meal (SBM)
Only source of dietary protein TiO2 (2.5 g/kg feed) was included
Wheat gluten (WG)
Rapeseed meal (RSM)
Porcine plasma protein meal
(PPM)
Transition (5 days)
Adaptation (14 days)
Dissection (2 days)
2.5 times the maintenance requirement for energy Twice a day in equal amount Feed: Water = 1:3
34
Parameters- protein digestion kinetics
Postprandial amino acids and peptides in
plasma
Determination of protein digestion kinetics
Digestibility and mean retention time
Peptide molecular weight distribution
35
Results- Total plasma AAs and peptides
3.00
4.00
5.00
6.00
7.00
8.00
9.00
10.00
-60 0 60 120 180 240 300 360 420 480
Plas
ma
amin
o ac
ids a
nd p
eptid
es (m
mol
e/l)
Postprandial time (min)
RSM
SBM
WG
Plasma
Diet (P < 0.01) Time (P < 0.01) Interaction (P < 0.01)
36
Results- Total plasma AAs and peptides Parameter estimates characterising the kinetics of postprandial concentration of amino acids (AAs) and peptides in systemic blood
ka (min-1) k10 (min-1) t1/2ka(min) t1/2k10(min)
RSM 0.0330 - 21 -
SBM 0.0046 0.0039 151 177
WG 0.0319 0.0053 22 130
PPM 0.0062 0.0056 113 124
ka (min-1) is the increase rate constant k10 (min-1) is the decrease rate constant t1/2ka (min) is the time where half of maximal concentration of AAs and peptides in systemic blood is reached t1/2k10 (min) is the time where half of maximal concentration of AAs and peptides in systemic blood is removed
37
Results- Area under the curve
0
200
400
600
800
1000
1200
120 240 480
Plas
ma
amin
o ac
ids a
nd p
eptid
es (
mm
ole/
l)
Postprandial time (min)
RSMSBM
a
b b b
a a
b b
b b
ab
a Diet (P < 0.001)
38
Parameters- protein digestion kinetics
Postprandial amino acids and peptides in
plasma
Determination of protein digestion kinetics
Digestibility and mean retention time
Peptide molecular weight distribution
0
50
100
150
200
250
300
350
400
450
Stomach 1st SI 2nd SI 3rd SI 4th SI Total SI
Mea
n re
tent
ion
time
(min
)
RSM
SBM
WG
PPM
39
Results- Mean retention time
ab ab b a
a ab
b b
Diet (P < 0.05)
a ab
b b
40
Results- CP Digestibility
0
10
20
30
40
50
60
70
80
90
100
1st SI 2nd SI 3rd SI 4th SI Total tract
Dige
stib
ility
coe
ffici
ent (
%)
RSM
SBM
WG
PPM
c
bc
a
ab
d
c
a b
a a
b
c
Diet (P < 0.001)
41
Results- Digestion curve
0.010.020.030.040.050.060.070.080.090.0
100.0
0 100 200 300 400 500 600
Dige
stib
ility
Coe
ffici
ent (
%)
Time (min)
SBM
0.010.020.030.040.050.060.070.080.090.0
100.0
0 100 200 300 400 500 600
Dige
stib
ility
Coe
ffici
ent (
%)
Time (min)
WG
0.010.020.030.040.050.060.070.080.090.0
100.0
0 100 200 300 400 500 600
Dige
stib
ility
Coe
ffici
ent (
%)
Time (min)
RSM
0.010.020.030.040.050.060.070.080.090.0
100.0
0 100 200 300 400 500 600Di
gest
ibili
ty C
oeffi
cien
t (%
) Time (min)
PPM
42
Results- Digestion curve
Parameter estimates characterising the kinetics of crude protein digestion in the small intestine of pigs Dmax (%) k (min-1) t1/2 (min) RSM 61 0.0136 63 SBM 78 0.0154 58 WG 94 0.0157 45 PPM 86 0.0338 20 Dmax (%) is potentially digestible CP (asymptote); k (min-1) is the rate constant; t1/2 (min) is the time where half of Dmax is reached.
43
Parameters- protein digestion kinetics
Postprandial amino acids and peptides in
plasma
Determination of protein digestion kinetics
Digestibility and mean retention time
Peptide molecular weight distribution
44
Results- Molecular weight distribution
0
100
200
300
400
500
600
Diet Stomach 1st SI 2nd SI 3rd SI 4th SI
Abso
rban
ce a
t 214
nm
SBM
0
100
200
300
400
500
600
Diet Stomach 1st SI 2nd SI 3rd SI 4th SI
Abso
rban
ce a
t 214
nm
RSM
0
100
200
300
400
500
600
Diet Stomach 1st SI 2nd SI 3rd SI 4th SI
Abso
rban
ce a
t 214
nm
WG
0
100
200
300
400
500
600
Diet Stomach 1st SI 2nd SI 3rd SI 4th SI
Abso
rban
ce a
t 214
nm
PPM
45
Conclusions • Protein sources differ in protein digestion kinetics up to
the end of the small intestine in pigs. Plasma AAs and peptides: RSM>WG>PPM>SBM
CP digestion rate up to ileum: PPM>WG>SBM>RSM
Molecular distribution: ??
WG and PPM: once major release of AAs and peptides SBM and RSM: continuous release of AAs and peptides • Nutrient synchronization increase utilization efficiency
of protein sources
Contact: [email protected]
TAKE HOME MESSAGE Protein digestion kinetics differs among protein sources in pigs different timing of postprandial appearance of amino acids and peptides in blood
Janne Beelen • Effects of a 12-week intervention with protein-enriched foods and
drinks on protein intake and physical performance of older patients during the first 6 months after hospital release
Effects of a 12-week intervention with protein-enriched foods and drinks on protein intake and physical performance of older patients during the first 6 months after hospitalization
Janne Beelen
Protein for Life
25 Oct 2016
Background
Protein requirements higher in older adults who are
acutely or chronically ill 1:
1.2 – 1.5 g protein/ kg body weight/ day
On average protein intake of older patients:
<0.9 g/kg/d
New strategies needed!
1 ESPEN Expert Group (Deutz, 2014) &
PROT-AGE Study Group (Bauer, 2013)
Study objective
To study the effectiveness of supplementing a hospital and
home menu with a protein-enriched assortment of
familiar foods and drinks
in reaching a protein intake of 1,2-1,5 g/kg/d
& in improving physical recovery during 12 weeks after
hospital stay in older patients
* Beelen et al. 2016, JNHA
Intervention products
Step 1: Interviews with elderly patients and dietitians
Step 2: Product
development
Step 3: Sensory evaluation
and Pilot study*
Study design
1 October 2014 – 1 October 2015
Pulmonary & Geriatric Medicine New patients: n = 860
Included: n= 147 Continued at home: n = 75
Exclusion criteria: - < 65 year - Expected hospital stay < 4 days - Delirious - Dietary protein restrictions - eGFR < 30 - Terminally ill
Exclusion criteria: - Living in nursing home - Cognitive impaired
Measurements at day before discharge and 2, 6, 12 and 24 weeks after hospitalization
Subjects: characteristics at admission
Controle (n=39) Intervention (n=36)
Age (mean ± SD) 77.2 ± 7.2 76.5 ± 6.7
Female/Male (n) 22/17 20/16
BMI (mean ± SD) 28.2 26.9
Medical diagnosis (n) - COPD, Asthma - Lung infection/inflammation - Other pulmonary diseases - Other infection (not lung) - Malaise or other
16 7 7 2 7
13 4 12 1 6
MUST score (n) - 0 - 1 - ≥2
33 2 4
26 3 7
Dietary assessment – Protein intake
• 24hr recall during home visit • Food record as a memory aid • Dutch Food Composition Table 2013
Results Protein intake
Control Cater with Care
Average intake during intervention period (wk. 2, 6, 12)
Control Intervention
Protein intake (g/kg/d) 1.0 ± 0.4 1.5 ± 0.6*
Reached intake of 1.2 g/kg/d 31% 72% * P < 0.01
Physical function: Short Physical Performance Battery
Balance (3 positions)
Gait speed (4 meter)
Chair rise time (5x)
Results Physical function
Control Cater with Care
Max score = 12 Linear Mixed Models:
No group or interaction effects
Same for other outcomes: Gait speed, Chair rise, Leg strength, Hand grip, Body weight, MNA, ADL, Physical Activity
Post-hoc analysis based on protein intake: below or above 1.2 g/kg/d
- Time (P<0.01) & Group effects (P=0.003)
- No interaction effect (P>0.05)
- Same letter on 2 time points: significant difference between these 2 time points within a group
Take Home message
Protein-rich and -enriched familiar foods and drinks successfully increased protein intake of older
patients, but did not improve physical recovery.
However, we do not rule out a beneficial role of these products in combination with exercise in older adults
that normally have low protein intakes.
More information: www.alliantievoeding.nl
Thank you! Any questions?
Acknowledgements
All participants
Cater with Care project partners
Astrid Doorduijn & Renske Geers
Dr. Emmelyne Vasse
Nancy Janssen
André Janse, MD.
Medical and Nursing staff, Hospital Gelderse Vallei
Cater with Care students
Dr. Nicole de Roos
Prof. Lisette de Groot
Lotte van der Zanden • Novel protein foods: what do elderly need, what do elderly want?
Imagine...
CHALLENGES IN MARKETING TO ELDERLY CONSUMERS Illustrated with the case of protein-enriched foods
Lotte van der Zanden, Ellen van Kleef, Rene de Wijk & Hans van Trijp October 26th, 2016 Wageningen University
Background
Source: IeC Analysis; Un population division
Source: UN World Population Prospects (2002) Eurostat Demographic Projections (2004)
Background
Background
Background
24%
76%
SuccessFailure
Background
Someone aged….
45+ (e.g. Weijters & Geuens, 2003)
50+ (e.g. Sudbury & Simcock, 2009)
55+ (e.g. Moschis & Friend, 2008)
60+ (e.g. Delaney & McCarthy, 2009)
64 (The Beatles, 1967)
65+ (e.g. Mattila, Karjaluoto & Pento, 2000)
Background
Source: www.projectcartoon.com
Background
This presentation
Methods
Methods
Focus groups
Methods
Narrative review
Methods
Segmentation
Methods
Field study
This presentation
Results (Focus groups)
“Not for us!”
Aged 58-93
Results (Focus groups)
“For pregnant
women, children, ill people, and . . .
for old people”
“Not for us!”
Aged 58-93
ID SUB ACC STA RES aCOM aDSG
cognitive age +- +- +- ++ ++ +- --
life course -- ++ +- ++ ++ -- --
time perspective +- -- -- +- ++ ++ ++
demographics ++ ++ ++ ++ -- -- --
general food beliefs -- ++ -- +- ++ +- +-
food choice motives +- ++ -- ++ +- ++ ++
attributes & benefits -- ++ -- +- ++ ++ ++
past purchase +- ++ +- ++ -- -- +-
Results
abbreviations: ID, identifiability; SUB, substantiality; ACC, accessibility; STA, stability; RES, responsiveness; aCOM, actionability for communication; aDSG, actionability for design
(Narrative review)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Mean age 67.0 66.5 66.6 67.3 66.4 69.7 67.1
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Mean age 67.0 66.5 66.6 67.3 66.4 69.7 66.3
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 7
Percentage 100% 31% 27% 21% 12% 6% 10%
R-squared 0.361 0.560 0.511 0.798 0.435 0.755 n.a.
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Percentage 100% 31% 27% 21% 12% 6% 3%
R-squared 0.361 0.560 0.511 0.798 0.435 0.755 0.385
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Mean age 67.0 66.5 66.6 67.3 66.4 69.7 66.3
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Percentage 100% 31% 27% 21% 12% 6% 3%
R-squared 0.361 0.560 0.511 0.798 0.435 0.755 0.385
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Mean age 67.0 66.5 66.6 67.3 66.4 69.7 66.3
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Percentage 100% 31% 27% 21% 12% 6% 3%
R-squared 0.361 0.560 0.511 0.798 0.435 0.755 0.385
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results
“Healthy products that we consume
frequently” (Focus groups)
(Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
“Healthy products that we consume
frequently” (Focus groups)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Healthiness 1.03 1.09** 0.85** 1.59** 1.09** 0.35* -.68*
Novelty -0.22 0.46** -.80** n.s. n.s. -1.56** -.61*
Meal type 0.40 0.72** 0.59** 0.43** -.42* n.s. -.67*
Results (Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Tailored formats
appropriateness 5.20 5.22 5.31 5.53 4.68 5.06 3.63
trial purchase 4.92 4.50 5.26 5.20 4.59 5.40 4.06
repeat purchase 4.18 3.31 5.15 4.57 3.38 4.95 2.44
Results
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
All formats appropriateness 3.45 3.06 3.65 3.38 4.86 4.00 3.74
trial purchase 3.17 2.83 3.23 3.37 3.33 3.51 3.41
repeat purchase 2.58 2.35 2.65 2.80 2.60 2.71 2.52
Measured on a scale from 1 to 7
(Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Tailored formats
appropriateness 5.20 5.22 5.31 5.53 4.68 5.06 3.63
trial purchase 4.92 4.50 5.26 5.20 4.59 5.40 4.06
repeat purchase 4.18 3.31 5.15 4.57 3.38 4.95 2.44
Results
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
All formats appropriateness 3.45 3.06 3.65 3.38 4.86 4.00 3.74
trial purchase 3.17 2.83 3.23 3.37 3.33 3.51 3.41
repeat purchase 2.58 2.35 2.65 2.80 2.60 2.71 2.52
Measured on a scale from 1 to 7 Or, roughly speaking, 1-2-3 4 5-6-7
(Segmentation)
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Tailored formats
appropriateness 5.20 5.22 5.31 5.53 4.68 5.06 n.a.
trial purchase 4.92 4.50 5.26 5.20 4.59 5.40 n.a.
repeat purchase 4.18 3.31 5.15 4.57 3.38 4.95 n.a.
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 7
All formats appropriateness 3.45 3.06 3.65 3.38 4.86 4.00 1.66
trial purchase 3.17 2.83 3.23 3.37 3.33 3.51 1.00
repeat purchase 2.58 2.35 2.65 2.80 2.60 2.71 1.01 Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
Tailored formats
appropriateness 5.20 5.22 5.31 5.53 4.68 5.06 3.63
trial purchase 4.92 4.50 5.26 5.20 4.59 5.40 4.06
repeat purchase 4.18 3.31 5.15 4.57 3.38 4.95 2.44
Results
Total Group 1 Group 2 Group 3 Group 4 Group 5 Group 6
All formats appropriateness 3.45 3.06 3.65 3.38 4.86 4.00 3.74
trial purchase 3.17 2.83 3.23 3.37 3.33 3.51 3.41
repeat purchase 2.58 2.35 2.65 2.80 2.60 2.71 2.52
Measured on a scale from 1 to 7 Or, roughly speaking, 1-2-3 4 5-6-7
(Segmentation)
Results (Field study)
“There is nothing extra in there, it is all just marketing.”
(Focus groups)
Control (N = 93) Prompt (N = 115) % fruit quark 6.5% 45.2%
Grams of protein 24.0 30.5
Would you like some
fruit quark?
Results (Segmentation)
iNspire
Discussion
• age is irrelevant
• protein is also irrelevant?
• the “average” elderly does exist, but...
• attractive ≠ purchaseable
• tailoring increases acceptance
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• protein is also irrelevant?
• the “average” elderly does exist, but...
• attractive ≠ purchaseable
• tailoring increases acceptance
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• like calcium is for bones
• the “average” elderly does exist, but...
• attractive ≠ purchaseable
• tailoring increases acceptance
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• like calcium is for bones
• focus groups VS market segmentation
• attractive ≠ purchaseable
• tailoring increases acceptance
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• like calcium is for bones
• focus groups VS market segmentation
• distinguish between measures of acceptance
• tailoring increases acceptance
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• like calcium is for bones
• focus groups VS market segmentation
• distinguish between measures of acceptance
• sometimes hiding it is better
• reaching rejecters likely the hardest
Discussion
• focus on preferences
• like calcium is for bones
• focus groups VS market segmentation
• distinguish between measures of acceptance
• sometimes hiding it is better
• inspire initial use and/or purchase
Segmentation van der Zanden, van Kleef, de Wijk & van Trijp (2015). Examining heterogeneity in elderly consumers’ preferences towards carriers for enriched food. Food Quality and Preference, 42, 130-138.
Field study van der Zanden, van Essen, van Kleef, de Wijk & van Trijp (2015). Using a verbal prompt to increase protein consumption in a hospital setting: A field study. International Journal of Behavioral Nutrition and Physical Activity 2015, 12, 110.
Focus groups van der Zanden, van Kleef, de Wijk & van Trijp (2014). Knowledge, perceptions and preferences of elderly regarding protein-enriched functional food. Appetite, 80, 16-22.
Narrative review van der Zanden, van Kleef, de Wijk & van Trijp (2014). Understanding heterogeneity among elderly consumers: An evaluation of segmentation approaches in the functional food market. Nutrition Research Reviews, 27, 159-171.
Thank you!
DAY 2, SESSION 6 Session 6
Topic: 2. Side-streams and insects Presentations available: • Marcel Hulst • Natasja Gianotten
105
Marcel Hulst • In vivo and in vitro study on the mode of action of spray dried plasma
when used as feed additive
IN VIVO AND IN VITRO STUDY ON THE MODE OF ACTION OF SPRAY DRIED PLASMA WHEN USED AS FEED ADDITIVE
HULST, MARCEL1,KAR, SOUMYA1,2 VAN VUURE, CARINE3; VAN DOREMALEN, ANKE3; HERES, LOURENS3; DE WIT, AGNES2; JANSMAN, ALFONS2; SMITS, MARI1,2
1 Wageningen UR Livestock Research, Wageningen, The Netherlands. 2 Host-Microbe Interactomics, Wageningen University, Wageningen, The Netherlands. 3 Darling Ingredients, Son, The Netherlands.
Spray dried (porcine) plasma (SDPP) is widely applied in weaning diets for piglets. Beneficial effects; shortens the period and reduce the severity of Post Weaning Diarrhea Objective > understanding in the mode of action of SDPP > can we use in vitro data to complement in-vivo data ? Approach; 1 -gene expression in ileum piglets after feeding a diet supplemented with SDPP 2- gene expression in porcine enterocyte cell line (IPEC-J2) exposed to SDPP 3 - overlapping in vivo-in vitro pathways/processes -“putative” effector genes/proteins expressed by enterocytes (IPEC-J2) 4- integration of in-vitro data (IPEC-J2) with in vivo data (piglets)
IN VIVO AND IN VITRO EXPERIMENT WITH SPRAY DRIED PORCINE PLASMA (SDPP)
enterocytes produce effector molecules (proteins/compounds) in response to changes in the composition of the luminal content of the ileum
Digestion of proteins in gastro-intestinal tract
Partially or Undigested Proteins (e.g. IgG)
Gene-expression analysis / microarray
• Measures activity of all genes (messenger-RNAs) in cells/tissues • 28.000 pig genes • Comparison treated versus control cells or tissues (ratio treated/non-treated; Fold Change)
mRNA +
1 2
3 4
5
60k expression profiles
“Which proteins are activated and which are
silenced “
List of response genes /Datamining / Functional analysis
Gene symbol FC S/M 2hr up/down
Gene symbol
FC ([M-S] vs [M]) up/down
CXCL2 53.23 up GPR158 -2.00 DOWN CXCL2 39.64 up RNF115 -2.00 DOWN IL1A 21.25 up TMEM151B -2.00 DOWN
CDK17 20.24 up SEC22C -2.00 DOWN CSF2 17.19 up HEATR4 -2.02 DOWN IL1A 13.88 up HYLS1 -2.03 DOWN
MAP3K8 13.41 up KLF13 -2.03 DOWN SLC2A5 13.28 up KLF13 -2.03 DOWN DDIT4 13.11 up FAM189A2 -2.03 DOWN CHAC1 12.75 up BEST2 -2.03 DOWN
TNFAIP3 12.11 up PIGV -2.03 DOWN SNAI1 8.48 up SYNGR3 -2.04 DOWN
IL8 7.42 up SIT1 -2.05 DOWN INHBA 6.41 up Pfn2 -2.05 DOWN CYP1A1 5.53 up CCNL1 -2.05 DOWN
IER3 5.51 up NOSTRIN -2.05 DOWN
Analysis list of IPEC-J2 response genes using bioinformatics programs
(FC: Fold change = ratio treated over control(s)
Pig Feeding experiment; Gene set enrichment analysis (GSEA), a statistical method to identify significantly enriched or depleted groups of genes associated with Gene Onthology terms (function).
IPEC-J2 experiment; differentially expressed genes with a Fold change (FC) of > 2.0 up- or down regulated (p< 0.05) and selection of up-regulated effector genes/proteins using phenotyping program.
Pathway >>
Associations with compounds
Enriched -pathways
-compounds
Pig feeding experiment with protein diets (Kar, Soumya et. al.)
Animal source -Spray dried porcine plasma protein (SDPP; Proglobulin Darling -ingredients) Plant source -Wheat gluten meal (WGM) -Rapeseed meal (RSM). -Soybean meal (SBM). Feeding experiment piglets -Six-week old pigs -3-weeks fed with experimental diets -Gene expression analysis of mucosal scrapings ileum. -Comparison results of SDPP diet to “plant protein” diets >> SDPP specific genes
Dietary protein level of 160 g/kg for protein sources. “No large differences in weight gain between groups”
IPEC-J2 in-vitro assay
LTB; Lymphotoxin Beta (cytokine) BTG2; B-Cell Translocation Gene 2 (cytokine) IL6; Interleukin 6 (cytokine) IL8; Interleukin 8 CXCL2; Chemokine (C-X-C Motif) Ligand 2 CSF2; Colony stimulating factor 2 (cytokine) enzymes metabolising chemical immune modulators! HPGD; Hydroxy-prostaglandin Dehydrogenase 15 (prostaglandines > regulation of inflammation/cytoprotection/platelet activation)
-Bioinformatics analysis of list with response genes >> pathways -Selection (secreted) effectors from lists of SDPP response genes.
Example immune-modulating effector genes
IL8 ↑ CSF2 ↑↑ IL6 ↓ CXCL2 ↑ LTB ↑ IL1A ↑ BTG2↓ HPGD ↑
IPEC-J2 cells
Exposure to 2 different concentration of SDPP
2 and 6h mRNA Microarray
Cultured IPEC-J2 cells grown for 6 -7 days form an enterocyte-like layer
Approach summarized; integration of in vitro – in vivo gene expression data
114
In-vivo pig experiment
enrichment analysis (GSEA); sets of
response genes specific for SDPP in the ileum (comparison to SBM,
WGM, RSM data).
Association of SDPP-specific
response- genes with
pathways and compounds
In-vitro IPEC-J2 experiment; SDPP
IPEC-J2 response genes with a FC>2 up-and
down (2 and 6h).
Participation of 28 IPEC-J2 EFFECTOR GENES in in-vivo
pathways
in-vitro IPEC-J2 experiment SDPP Genes with FC>2.
up-regulated; EFFECTOR GENES >> secreted genes
coding for cytokines-chemokines-hormones-growth factors- immune regulators-proteases and
inhibitors.
Association of IPEC-J2
response genes with
pathways and compounds
Overlapping compounds, with IPEC-J2 effector
Overlapping pathways, with IPEC-J2 effector
115
Results (1) IPEC-J2 effector genes participating in in vivo pathways/processes
28 effector genes; up-regulated expression in response to SDPP (FC>2) CASP7 apoptosis inducer (non-secreted) TNFAIP3 apoptosis inhibitor CXCL10 chemokine CXCL2 chemokine IL8 chemokine IL1A cytokine CSF2 cytokine IL1RN cytokine VEGFA endothelial growth factor EDN3 endothelial growth factor NRG1 growth factor INHBA hormone/growth factor NPPB hormone/growth factor HBEGF smooth muscle cell proliferation/growth factor ITGA2 focal adhesion MMP13 Metalloendopeptidase NGF Metalloendopeptidase Inhibitor Activity/growth factor TIMP3 Metalloendopeptidase Inhibitor Activity PLAT Serine-type Endopeptidase activity F3 Serine-type Endopeptidase activity COL7A1 Serine-type Endopeptidase Inhibitor Activity CRISPLD2 Serine-type Endopeptidase Inhibitor Activity SERPINE1 Serine-type Endopeptidase Inhibitor Activity SLPI Serine-type Endopeptidase Inhibitor Activity PRDM1 Transcription repressor / Drives the maturation of B-lymphocytes (non-secreted) HMOX1 anti-inflammatory proteins whenever oxidation injury takes places PPARG steroid/thyroid hormone coactivator receptor (non-secreted) PTGS1 Prostaglandin synthesis (Prostaglandins; secreted)
Results (2) IPEC-J2 effector genes participating in in vivo pathways
term Overlapping in-vivo/in-vitro pathways (SDPP specific genes) IPECJ2-effectors part of in-vivo pathway
angiogenesis Angiogenesis (CST) HMOX1, TIMP3 apoptosis Apoptotic Pathways PPARG, ITGA2, NRG1, HBEGF, NGF, TIMP3, CASP7
Ca2+ Ca-dependent Events NRG1, SERPINE1, VEGFA, F3 ECM Cell Adhesion / ECM Remodeling MMP13, PLAT, SERPINE1, HBEGF, TIMP3, IL8, ECM Endochondral Ossification MMP13, PLAT, VEGFA, TIMP3 ECM Integrin Cell Surface Interactions ITGA2 IL2 IL-2 Pathway IL1A, NRG1, HBEGF
NFKB NF-kappa B Signaling Pathway HMOX1, PRDM1, IL1RN, PTGS1, CXCL10, TNFAIP3 Antigen recognition Nucleotide-binding Domain, LRR Containing Receptor (NOD) PRDM1,TNFAIP3, CASP7, CXCL2, IL8 Antigen recognition Toll-like Receptor Signaling Pathway SERPINE1, CXCL10, IL8
TGF TGF-beta Signaling Pathway INHBA B-cell B Cell Receptor Signaling Pathway (KEGG) IL1A, CSF2
Results (3) Common compounds, association with IPEC-J2 effectors (most important)
TERM – Matching COMPOUND – function (Enterocyte effector genes)
Angiogenesis -VEGF - endothelial growth factors (VEGFA, EDN3, HMOX1, TIMP3)
Coagulation - Heparin - Serine-type Endopeptidase (inhibitor) activity (PLAT, F3, SERPINE1)
Inflammation – Dexamethasone, anti-inflammatory drug – (cytokines, chemokines, HMOX1)
ECM - Cysteine - Metalloendopeptidase (Inhibitor) activity (e.g. MMP13, TIMP3, NGF)
ECM-Serine -Serine-type Endopeptidase activity/Inhibitor (SLPI, CRISPLD2, COL7A1, SERPINE1)
Immune modulation- Pge2 - Prostaglandin synthesis (PTGS1, HPGD )
Overlapping compounds, with IPEC-J2
effector Pge2 -Inflammation -cytoprotection -platelet activation
-Overlap in SDPP-induced pathways/processes between the ileum and in-vitro cultured enterocytes. -Short-term response in IPEC-J2 enterocytes identified a set of effector genes/proteins providing insight in how biological processes imposed by a SDPP-rich diet in the ileum are regulated. > Remodeling and repair of the extracellular matrix. > Stimulation of the innate (via Toll- and NOD-like receptors) and adaptive (B-cell) immune system. Conclusion Insight in the mode of action how the protein-rich additive SDPP may support homeostasis in the intestine of piglets.
Summary and Conclusion
Natasja Gianotten • Sustainable protein production using lesser mealworms
SUSTAINABLE PROTEIN PRODUCTION USING LESSER MEALWORMS
PROTEIN FOR LIFE CONFERENCE
Natasja Gianotten Proti-Farm R&D BV
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60% increase 1+ Billion 840M people
18% of all GHG 70% of world’s 25% of world’s
Children suffer from malnutrition
Emissions come from livestock farming
Arable land is used for livestock farming
Don’t have enough to eat today
People without fresh water in 2050
Of meat consumption till 2050
GLOBAL CHALLENGES
More iron than spinach
More calcium
than milk
Natural vitamins & minerals Healthy
fats
High protein content
All nine essential amino
acids
WHY INSECTS - NUTRITIONAL
WILD CAUGHT INSECTS
FARMED INSECTS
Mealworm Tenebrio Molitor 1-2
cm
Giant mealworm Zophobas Morio 2,5-
3cm
Lesser Mealworm Alphitobius diaperinus 0,7 cm
PROTI-FARM’S MEALWORMS
Proti-Farm production facilities The Netherlands Fully mechanised and automated Controlled environment Vertical, sustainable farming
LARGE SCALE UP PRODUCTION OF LMW
Low Feed Conversion Ratio
Limited GHG Impact
Low Land Use
Low Water Use
Feed / raw material choice
INSECTS - SUSTAINABILITY
Feed /raw materials / FCR
Make use of by-products from food industry or food grade ‘rest streams’
Improve FCR by feeding method,
feed quality
High productivity => good quality feed
INSECTS - SUSTAINABILITY