nutrients

Article
Protein from Meat or Vegetable Sources in Meals
Matched for Fiber Content has Similar Effects on
Subjective Appetite Sensations and Energy
Intake—A Randomized Acute Cross-Over Meal
Test Study
Lone V. Nielsen *, Marlene D. Kristensen, Lars Klingenberg, Christian Ritz ID
, Anita Belza,
Arne Astrup ID and Anne Raben ID
Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Rolighedsvej 30,
DK-1958 Frederiksberg, Denmark; [email protected] (M.D.K.); [email protected] (L.K.);
[email protected] (C.R.); [email protected] (A.B.); [email protected] (A.A.); [email protected] (A.R.)
* Correspondence: [email protected]; Tel.: + 45-3532-0552

Received: 25 September 2017; Accepted: 11 January 2018; Published: 16 January 2018

Abstract: Higher-protein meals decrease hunger and increase satiety compared to lower-protein
meals. However, no consensus exists about the different effects of animal and vegetable proteins on
appetite. We investigated how a meal based on vegetable protein (fava beans/split peas) affected
ad libitum energy intake and appetite sensations, compared to macronutrient-balanced, iso-caloric
meals based on animal protein (veal/pork or eggs). Thirty-five healthy men were enrolled in this
acute cross-over study. On each test day, participants were presented with one of four test meals
(~3550 kilojoules (kJ) 19% of energy from protein), based on fava beans/split peas (28.5 g fiber),
pork/veal or eggs supplemented with pea fiber to control for fiber content (28.5 g fiber), or eggs
without supplementation of fiber (6.0 g fiber). Subjective appetite sensations were recorded at
baseline and every half hour until the ad libitum meal three hours later. There were no differences
in ad libitum energy intake across test meals (p > 0.05). Further, no differences were found across
meals for hunger, satiety, fullness, prospective food consumption, or composite appetite score
(all p > 0.05). Iso-caloric, macronutrient-balanced, fiber-matched meals based on vegetable protein
(fava beans/split peas) or animal protein (veal/pork or eggs) had similar effects on ad libitum energy
intake and appetite sensations.

Keywords: animal protein; fava beans; split pea; veal; pork; bean; satiety; egg

1. Introduction
In recent decades, high protein diets have attracted considerable attention and substantial
evidence now supports improved weight loss and weight loss maintenance after consumption of
high-protein diets [1]. These effects can be, at least partly, attributed to the highly satiating and
thermogenic effect of proteins [2,3].
Health authorities recommend an increase in dietary protein from plant-based sources, at the
expense of animal protein, since plant-based proteins sources have low energy density, high fiber
content, and their production is associated with a lower emission of greenhouse gases [4,5]. Lately,
several studies have investigated the different effects of proteins from plant and animal sources on
appetite and energy intake [6–8]. Kehlet et al. found no difference in appetite sensations and energy
intake when soy-based vegetable patties were compared to fiber-matched meatballs in an acute meal
test study [6]. Douglas et al. also found no differences in hunger, fullness, or energy intake when

Nutrients 2018, 10, 96; doi:10.3390/nu10010096 www.mdpi.com/journal/nutrients

Nutrients 2018, 10, 96 2 of 11

comparing a high-protein soy-based lunch with a macronutrient- and fiber-matched high-protein

beef-based lunch [7]. However, they investigated meals with a fairly large protein content (33 E%),
which could have masked potential differences between the protein sources. We have previously shown
that a meal with vegetable protein from fava beans and split peas increased satiety and decreased
hunger and ad libitum energy intake compared to a meal with animal protein from pork and veal [8].
That study was, however, not able to demonstrate that vegetable protein from legumes per se was
more satiating than animal protein, as the study allowed natural differences in dietary fiber content
across the meals [8].
Only a few studies have investigated how egg affects appetite compared to other protein sources.
Pal and Ellis found whey protein to be superior in stimulating satiety compared to tuna, egg, and turkey
when served as a macronutrient-balanced liquid breakfast meal. Also, tuna stimulated satiety to a
greater extent than egg or turkey [9]. Additionally, Anderson et al. observed a higher energy intake
after a liquid preload of egg albumin compared to a preload with soy or whey protein [10]. However,
it is not evident if the effects of protein from eggs are similar when ingested as solid foods in whole
meals [11].
Studies that compare animal and vegetable protein sources using whole meals, controlling for
macronutrient content and fiber, are limited. Therefore, the aim of the present study was to investigate if
a meal based on vegetable protein from fava beans and split peas would reduce ad libitum energy intake
and subjective appetite sensations compared to iso-caloric, macronutrient-balanced, fiber-matched
meals based on animal protein from pork and veal or eggs. The role of a similar egg meal without
supplemented fiber was also investigated.

2. Materials and Methods

2.1. Subjects
Healthy, weight stable (maximum ± 3 kg in the previous two months), non-smoking men,
aged 18–50 years, with a body mass index (BMI) of 18.5–30.0 kg/m2 , were recruited for the study.
Exclusion criteria included: mental disorders, metabolic diseases, systematic or/and chronic infections,
use of medication that is considered to affect appetite, food allergies of relevance to the test meals,
vegetarians, vigorous physical activity of more than 10 h per week, current or previous drug abuse,
alcohol intake above 14 units per week, or simultaneous participation in other studies. The participants
were individually interviewed reading the inclusion and exclusion criteria at a screening visit,
where they also completed the three-factor eating questionnaire [12]. Height was measured on a
wall-mounted stadiometer with 0.5-centimeter accuracy (HeightTronic235, Quick Medical, Issaquah,
WA, United States of America). Body weight was recorded to the nearest 0.05 kg on a decimal scale
(LindelTronic 8000, Samhall Lavi AB, Kristianstad, Sweden).
All subjects gave their written informed consent for inclusion before they participated in the
study. The study was conducted in accordance with the Declaration of Helsinki of 1975, revised in
1983, and was registered at clinicaltrials.gov (NCT01616251).

2.2. Experimental Design

This study was performed at the Department of Nutrition, Exercise and Sports, University of
Copenhagen, Denmark between August and December 2012, and was conducted as a single-blinded
randomized, four-condition, cross-over meal test study. On the four test days, the participants
arrived in the laboratory at the same time in the morning (at 8:00 a.m. or 8:30 a.m.). Upon arrival,
participants were required to attempt to void and body weight was recorded. After a 10-min rest,
the first visual analogue scales (VAS) were provided to assess subjective appetite sensations in the
fasting state. Participants were then served one of the four test meals (time 0), which they ingested
during the next 15 min. Subsequently, the participants filled in a VAS on subjective appetite sensations,
and for the palatability of the test meal. Assessment of subjective appetite sensations was repeated at
Nutrients 2018, 10, 96 3 of 11

30, 60, 90, 120, 150, and 180 min after consumption of the meal. At 180 min, the participant completed
an additional VAS regarding gastro-intestinal feelings. Three hours after the test meal an ad libitum
lunch meal was served. After the ad libitum meal, the participants were asked to complete the last
VAS regarding appetite sensations and palatability. To validate possible compensatory energy intake,
participants were instructed to complete a weighed food record from the time they were discharged
from the laboratory until midnight. On the first test day, the participants were carefully instructed on
how to fill in the records. A timeline of the test day measurements is shown in Table 1. Participants were
placed in rooms with a maximum of three participants in each, and shielding was placed so they
were not able to see each other. During the test day, participants were told to remain awake and were
not allowed to talk or communicate with each other. They were not allowed to use mobile phones,
the Internet, listen to music, watch films, or play computer games. They were allowed to read and
work on their laptops, unless it was related to food or health.

Table 1. Timeline of the measurements on each test day.

Time from Baseline (Minutes)

−5 0 15 30 60 90 120 150 180 ~200 ~200 to ~920
Test meal x
Ad libitum meal x
Subjective appetite
x x x x x x x x x
sensations
Palatability x x
Gastro-intestinal feelings x
Compensatory eating x

2.3. Standardization
Participants were instructed to eat a standardized 4 MJ dinner (a paprika dish with pork and
rice: 17 E% protein, 50 E% carbohydrate and 33 E% fat), provided as a frozen product beforehand,
between 7.00 p.m. and 8.00 p.m. on the evening prior to the test days and to fast from 8.00 p.m.
onward. During the fast, participants were allowed to drink 500 mL tap water, with a maximum of
250 mL allowed on the morning of the test days. Furthermore, the participants were told to abstain
from alcohol, medicine, and vigorous physical activity 24 h prior to the test days. The test days were
separated by a washout period of seven or more days.

2.4. Test Meals

Test meals were served as breakfast meals and consisted of (1) patties with beans served with
mashed split peas; (2) patties with minced pork/veal and pea fiber served with fiber-supplemented
mashed potatoes; (3) patties with eggs and pea fiber served with fiber-supplemented mashed potatoes;
and (4) patties with eggs served with non-fiber-supplemented mashed potatoes. Additionally, all meals
were served with ketchup and water. Water loss during preparation was higher for the bean patties,
and therefore extra water was served together with the bean and pea meal (300 mL and 333 mL,
respectively). The participants were required to consume the entire meal and to drink the water during
the meal. The four test meals were iso-caloric (~3550 kJ) and had a similar macronutrient distribution
of 19 E% protein, 53 E% carbohydrate, and 28 E% fat (Table 2). Pictures of the test meals are shown
in Figure S1.
Nutrients 2018, 10, 96 4 of 11

Table 2. Recipes and macronutrient composition of the four test meals.

Meat/Fiber (g) Bean/Pea (g) Egg/Fiber (g) Egg (g)

Pork/veal 4% 135 Fava Beans 100 Egg, whole 211 Egg, whole 211
Onion 10 Split peas 90 Egg white 40 Egg white 40
Potato 264 Onion 10 Onion 10 Onion 10
Potato flour 38.5 Water 445.5 Potato 305 Potato 338
Water 164 Rapeseed oil 10 Potato flour 41 Potato flour 50
Pea fibers 26 Butter 17 Water 17 Water 0
Rapeseed oil 9 Wheat flour 4.5 Pea fibers 26.5 Cream, 9% 33
Butter 13.2 Vinegar 5 Cream, 9% 33 Tomato ketchup 20
Rasp 20 Tomato ketchup 20 Tomato ketchup 20 Parsley 4
Tomato ketchup 20 Parsley 4 Parsley 4 Garlic 2
Parsley 4 Garlic 2 Garlic 2 Salt 3.3
Garlic 2 Salt 3.3 Salt 3.3 Pepper 0.02
Salt 3.3 Pepper 0.02 Pepper 0.02
Pepper 0.02
Energy (kJ) 3564.8 Energy (kJ) 3567.1 Energy (kJ) 3565.6 Energy (kJ) 3569.0
Weight (g) 709 Weight (g) 711 Weight (g) 713 Weight (g) 711
Density (kJ/g) 5.0 Density (kJ/g) 5.0 Density (kJ/g) 5.0 Density (kJ/g) 5.0
Protein (E%) 18.5 Protein (E%) 18.7 Protein (E%) 18.9 Protein (E%) 18.7
CHO (E%) 53.5 CHO (E%) 53.3 CHO (E%) 53.0 CHO (E%) 53.3
Fat (E%) 28.0 Fat (E%) 28.0 Fat (E%) 28.1 Fat (E%) 28.0
Fiber (g) 28.8 Fiber (g) 28.4 Fiber (g) 28.5 Fiber (g) 6.0
E%: energy percent; kJ: kilojoule; CHO: carbohydrate.

To obtain equal fiber content in the three fiber-matched meals, pea fiber (Fibradan, DLG Food
A/S, Copenhagen, Denmark) was added to the two fiber-supplemented meals. The pea fiber was
produced from the shell of dry yellow and green peas. The product contained cellulose (48.5%),
hemicellulose (18%), pectin (14%), lignin (2%), and other compounds (17.5%), total dietary fiber
content in the product was 90% (w/w) [13]. The physiochemical properties of the pea fiber have
previously been described [13].

2.5. Ad Libitum Meal

The ad libitum lunch meal was pasta bolognaise (556 kJ per 100 g; 15 E% protein,
55 E% carbohydrate, and 30 E% fat). Participants were provided with a pot with 1440 g pasta
bolognaise together with 300 mL water and served themselves. They were instructed to eat until
they felt comfortably satiated. The pot was weighed before the participants were served the meal.
Afterward, the pot was weighed again and the amount ingested was calculated.

2.6. Measurements of Subjective Appetite Sensations and Palatability

Subjective appetite sensations, well-being, and palatability of the meals were assessed using a
100-mm VAS. At each end of the line, the most positive or most negative ratings were anchored [14].
The VAS questions were combined in a small booklet showing one question at a time. The participant
received a total of nine VAS booklets on appetite and well-being during the test day. All booklets
included questions regarding satiety, fullness, prospective food consumption (PFC), hunger, thirst,
well-being, and subjective sensory-specific desire for sweet, fat, salt, and meat or fish. The composite
appetite score was calculated as [satiety + fullness + (100 − PFC) + (100 − hunger)]/4 [15]. The VAS
questions on appetite included: (1) How hungry do you feel? Responses could range from ‘I am not
hungry at all’ to ‘as hungry as I have ever felt’; (2) How full do you feel? Responses could range
from ‘not full at all’ to ‘totally full’; (3) How satisfied do you feel? Responses could range from ‘I am
completely empty’ to ‘I cannot eat another bite’; (4) How much do you think you could eat right now?
Responses could range from ‘nothing at all’ to ‘a lot’. Well-being was assessed with the question:
how comfortable do you feel? Responses could range from ‘really bad’ to ‘really good’. Palatability of
the meals was assessed immediately after the participants finished the test meal and again after the ad
libitum meal. The VAS question on palatability was: palatability ‘bad’ to ‘good’.
Nutrients 2018, 10, 96 5 of 11

2.7. Gastro-Intestinal Feelings

The VAS booklet used to assess gastrointestinal feelings included eight yes/no questions. Here,
participants answered whether they had experienced acid reflux, nausea, stomach pain, flatulence,
heartburn, bloating, rumbling, or diarrhea. If the participants answered yes, they had to rate the
intensity of the symptom on a 100 mm scale.

2.8. Compensatory Eating

Food records, used to validate possible compensatory eating, were entered into the Danish dietary
software program (Dankost 3000® version 07.11.03, Dansk Catering Center, Copenhagen, Denmark)
and energy intake during the remainder of each test day was calculated.

2.9. Blinding and Randomization

The different meals were coded with a number from 1 to 4. Only the kitchen staff, who prepared
the test meals, knew this combination. The kitchen staff provided the test meals to the investigator
with an opaque lid and the investigator was not aware of which meal the participant received on
the different test days. Before the study was initiated, a list of combinations of the numbers 1–4 was
generated. When participants were found eligible for the study, they were assigned the next available
combination of meals by the study coordinator.

2.10. Statistical Power

The primary endpoint was ad libitum energy intake three hours after the test meals. The sample
size calculation was based on results from a previous study [8], which showed that 30 participants had
to complete the study to detect a mean difference in ad libitum energy intake of 400 kJ three hours
after the test meal, with a standard deviation of 750 kJ (α = 0.05, β = 0.8).

2.11. Statistical Analysis

All statistical analyses were performed using R version 3.1.2 (R Core Team, 2016, Vienna, Austria).
Baseline characteristics are presented as mean ± standard deviation (SD). Ad libitum energy intake,
palatability, incremental area under the curve (iAUC), and incremental area over the curve (iAOC) were
analyzed using linear mixed models with meal as fixed effect and subject as random effect. The models
were adjusted for age, BMI, meal order, and visit number. As the test meals had a fixed energy content,
an additional model, adjusted for estimated daily energy requirement based on the Harris–Benedict
equation and an estimated physical activity level of 1.7, was tested for ad libitum energy intake [16].
iAUC and iAOC were calculated using the trapezoidal method. Repeated measurements of subjective
appetite sensations were analyzed using linear mixed models, which included a time–meal interaction.
The models were adjusted for visit number, age, BMI, fasting value on the actual test day, and meal
order. Additional analyses, adjusted for estimated daily energy requirement, were conducted for satiety,
hunger, fullness, and PFC. Overall subject and within-visit subject differences were modeled through
random effects. Serial correlation between repeated measurements for the same subject within each
visit was modeled, assuming a spatial Gaussian correlation structure with decreasing correlation over
time. Post hoc tests were conducted as model-based pairwise comparisons adjusted for multiple testing.
Table S1 shows pairwise comparisons for each of the outcomes. Multiplicity adjustment of p-values was
based on the single-step method [17]. Results are presented as mean ± standard error (SE); graphs are
based on raw data presented as mean ± standard error of mean (SEM). Additional sensitivity analyses
with adjustment for palatability were conducted. For all models, assumptions about normality and
homogeneity of variance were checked graphically using residual plots and normal probability plots.
Differences in occurrence of gastrointestinal feelings between meals were evaluated using Fisher’s
exact test. p-values < 0.05 were considered significant.
Nutrients 2018, 10, 96 6 of 11

plots. Differences in occurrence of gastrointestinal feelings between meals were evaluated using
Nutrients 2018, 10, 96 6 of 11
Fisher’s exact test. p-values < 0.05 were considered significant.

3. Results
3. Results

3.1.3.1.
Subjects
Subjects
Thirty-nine potential
Thirty-nine male
potential participants
male attended
participants attended a screening visit
a screening at the
visit university.
at the Thirty-five
university. Thirty-five
participants were
participants found
were eligible
found and
eligible recruited
and to to
recruited thethe
study (Table
study 3).3).
(Table

Table 3. Baseline
Table characteristic
3. Baseline of the
characteristic participants.
of the participants.

Mean ± SD Range
Mean ± SD Range
Age (year) 26.5 ± 5.5 19–39
Age (year) 26.5 ± 5.5 19–39
BMI, kg/m2 2 23.3 ± 1.9 22.2–27.8
BMI, kg/m 23.3 ± 1.9 22.2–27.8
Restraint 1
Restraint 1 5.67
5.67± ±
2.87
2.87 0–110–11
Disinhibition
Disinhibition11 5.03
5.03± ±
2.12
2.12 2–132–13
Hunger1 1
Hunger 4.11± ±
4.11 2.57
2.57 1–111–11
1 Based on the three-factor eating questionnaire [12]. SD, standard deviation; BMI, body mass index.
1 Based on the three-factor eating questionnaire [12]. SD, standard deviation; BMI, body mass index.

OneOneparticipant dropped
participant dropped outouton on
thethe
first experimental
first experimental day, as as
day, hehe
waswasunable to to
unable consume
consume thethe
testtest
meal. Another
meal. Anotherparticipant
participantreceived
receivedthethe
samesametesttest
meal twice
meal duedue
twice to an experimental
to an experimentalerror. Thus,
error. Thus,
33 33
participants received
participants allall
received four test
four meals
test meals(Figure
(Figure 1).1).

Figure 1. Participant
Figure flowchart.
1. Participant flowchart.

3.2.3.2.
AdAdLibitum Energy
Libitum Intake
Energy Intake
NoNo
differences
differenceswere found
were foundin in
energy
energyintake
intakeat at
thethe
adadlibitum
libitummeal
mealacross test
across meals:
test meat/fiber
meals: meat/fiber
2636
2636 ± 200 kJ (630 ± 48 kcal), egg 2771 ± 201 kJ (662 ± 48 kcal), bean/pea 2608 ± 194 kJ (62337
± 200 kJ (630 ± 48 kcal), egg 2771 ± 201 kJ (662 ± 48 kcal), bean/pea 2608 ± 194 kJ (623 ± ± kcal),
37 kcal),
egg/fiber 2513
egg/fiber ± 203
2513 kJ (600
± 203 ± 48±kcal),
kJ (600 p > p0.05
48 kcal), (Figure
> 0.05 2). 2).
(Figure TheThelevel of significance
level of significancediddid
notnot
change
change
when estimated daily energy requirement was included as a covariate
when estimated daily energy requirement was included as a covariate (p > 0.05). (p > 0.05).
Nutrients 2018, 10, 96 7 of 11
Nutrients 2018, 10, 96 7 of 11

3000

(kJ)
intake (kJ)
2000

Energy intake
Meat/Fibers
Egg

Energy
1000
Bean/Pea
Egg/Fibres
0

Figure
Figure 2. Ad
Ad libitum
libitum energy
energy intake
intake three
three hours
hours after
after the
the participants
participants received
received the
the test
test meals,
meals,
analyzed
analyzed using
using a mixed linear model including meal as a fixed effect.

3.3.
3.3. Subjective
Subjective Appetite
Appetite Sensations,
Sensations, Sensory-Specific
Sensory-Specific Appetite Scores, and
Appetite Scores, and Palatability
Palatability
Satiety, hunger, fullness,
Satiety, hunger, fullness, and
and desire
desire toto eat
eat meat
meat oror fish,
fish, something
something salty, sweet, or
salty, sweet, or fatty
fatty did
did not
not
differ
differ after
after the
thead
adlibitum
libitummeal
mealacross
acrosstest
testmeals.
meals.However,
However, thethe
PFC rating
PFC waswas
rating higher following
higher the
following
bean/pea meal compared to the egg/fiber meal (16.1 ± 2.2 mm vs. 11.4
the bean/pea meal compared to the egg/fiber meal (16.1 ± 2.2 mm vs. 11.4 ± 2.3 mm, p < 0.05). ± 2.3 mm, p < 0.05). No
differences
No differences werewere
foundfound
in palatability of the ad
in palatability of libitum
the ad meal libitumafter the after
meal four different
the fourtest meals (p
different >
test
0.05).
meals (p > 0.05).
Fasting
Fasting values
values of
of hunger,
hunger, satiety,
satiety, fullness,
fullness, and
and PFC
PFC werewere not
not different
different across
across the
the test
test meals
meals (all,
(all,
pp >> 0.05).
0.05). No
No time
time ** meal
meal interactions
interactions or or meal
meal effects
effects were
were found
found for
for the
the postprandial
postprandial changes
changes in in
hunger, satiety, fullness, or PFC across test meals (all p > 0.05), neither when analyzed
hunger, satiety, fullness, or PFC across test meals (all p > 0.05), neither when analyzed separately nor separately nor
when combined into
when combined the composite
into the composite score
score (p (p >
> 0.05).
0.05). The
The level
level of
of significance
significance did did not
not change
change when
when
estimated daily energy requirement was included as a covariate (p > 0.05).
estimated daily energy requirement was included as a covariate (p > 0.05). These results remainedThese results remained
unchanged
unchanged when when the
the appetite
appetite ratings
ratings were
were analyzed
analyzed as as iAUC
iAUC or or iAOC
iAOC (all
(all pp >
> 0.05)
0.05) (Figure
(Figure 3).
3).

100 Meat/Fibers
Egg
80 Bean/Pea 10000
10000
(mm)
Satiety (mm)

Egg/Fibres 8000
8000
60
satiety
iAUC satiety

6000
Satiety

6000 Meat/Fibers
40
Egg
iAUC

4000
4000
Bean/Pea
20 Egg/Fibers
2000
2000
0
00
-5 15 30 60 90 120 150 180
Relative time (min)
(b)
(a)

100
Meat/Fibers
Egg 10000
10000
80
Bean/Pea
(mm)
Hunger (mm)

8000
8000
60 Egg/Fibres
hunger
iAOC hunger

6000
6000
Hunger

40 Meat/Fibers
Egg
iAOC

4000
4000
Bean/Pea
20 2000
2000 Egg/Fibers
0 00
-5 15 30 60 90 120 150 180
Relative time (min)
(c) (d)

Figure 3. Mean
Figure 3. Mean unadjusted
unadjusted three-hour
three-hour changes
changes in
in (a)
(a) satiety,
satiety, (c)
(c) hunger,
hunger, and
and (b) corresponding
(b) corresponding
incremental areaunder
incremental area under
thethe
curvecurve
(iAUC)(iAUC) and
and (d) (d) incremental
incremental area overarea over (iAOC),
the curve the curve (iAOC),
respectively.
respectively. Data are presented as mean ± SEM. Both as repeated measures, analyzed
Data are presented as mean ± SEM. Both as repeated measures, analyzed using mixed linear models using mixed
linear models including a time * meal interaction, and iAUC or iAOC, analyzed using
including a time * meal interaction, and iAUC or iAOC, analyzed using mixed linear models including mixed linear
models including meal as a fixed effect. No differences were found in satiety and
meal as a fixed effect. No differences were found in satiety and hunger across meals (all p > 0.05).hunger across
meals (all p > 0.05).
Nutrients 2018, 10, 96 8 of 11

No differences were found in the desire to eat something salty, sweet, or meat or fish across the
test meals (all p > 0.05). However, a significant meal * time interaction was found for the desire to
eat something fatty (p < 0.05). Pairwise comparisons, however, did not reveal differences across test
meals at any time point (all p > 0.05). When analyzing the sensory-specific appetite scores as iAUC,
no differences were found across the test meals (all p > 0.05). In addition, thirst and well-being were not
different across the meals (p > 0.05) and no differences existed in gastrointestinal feelings (all p > 0.05).
Palatability was higher for the meat/fiber meal compared to the bean/pea meal (61.3 ± 6.1 mm
vs. 39.7 ± 5.7 mm, p < 0.001) and the egg/fiber meal (61.3 ± 6.1 mm vs. 45.8 ± 6.2 mm, p < 0.01)
and for the egg meal compared to the bean/pea meal (54.6 ± 6.1 mm vs. 39.7 ± 5.7 mm, p < 0.05).
After adjusting the subjective appetite ratings for palatability of the test meals, an overall effect of
meal was seen on hunger (p < 0.05). Pairwise comparisons revealed that hunger decreased after the
egg/fiber meal compared to the meat/fiber meal (p < 0.05).

3.4. Compensatory Eating

The completion rate of the food records was 99.2% (134 of 135 potential records were received).
No differences were found in energy intake across test meals during the rest of the test day (meat/fiber:
6331 ± 604 kJ (1512 ± 144 kcal); egg: 6009 ± 550 kJ (1435 ± 131 kcal); bean/pea: 6005 ± 567 kJ
(1435 ± 135 kcal); and egg/fiber: 5806 ± 619 kJ (1387 ± 148 kcal), (p > 0.05).

4. Discussion
In this present study, we did not observe any differences in ad libitum energy intake or
appetite sensations between test meals based on vegetable protein compared to fiber-supplemented,
macronutrient-balanced, iso-caloric meals based on pork/veal or eggs. Furthermore, no differences
were observed when compared to a non-fiber-supplemented egg meal.
The majority of the previously conducted studies that investigated the effect of vegetable
protein compared to animal protein on appetite and energy intake used soy as the vegetable protein
source [6,7,18,19]. Douglas et al. investigated how a high-protein lunch with beef affected appetite
sensations and energy intake compared to a macronutrient- and fiber-matched high-protein soy
lunch [7]. In agreement with our study, no differences in appetite sensations or energy intake between
the vegetable and animal protein meal were found. A study by Kehlet et al. supported these findings,
as they found a similar effect of fiber-matched meals based on pork or soy on appetite sensations and
energy intake [6]. Additionally, a randomized acute meal study by Tan et al. found no differences in
satiety levels across iso-caloric high-protein meals with either meat (lean beef and ham), dairy (low-fat
milk, cheese, and yogurt), or soy as protein sources [20]. Lang et al. evaluated different protein sources
and found no differences in appetite sensations or ad libitum energy intake during the subsequent
dinner, after energy-fixed lunch test meals with either, casein, gelatin, or soy [18]. The results from the
latter two studies should be interpreted with caution. Tan et al. did not include information regarding
fiber whereas the soy meal in the study by Lang et al. contained a higher amount of fiber than the
casein and gelatin meal [18,20]. Contrary to these findings, preloads of pasta with tofu or mycoprotein
have been observed to reduce energy intake 20 min after consumption compared to a preload of pasta
with chicken [19]. These findings are, however, confounded by higher energy content in the meals with
tofu and mycoprotein compared to the meal with chicken and higher fiber content in the tofu meal.
Only a limited number of studies have investigated how bean and pea protein affect appetite
sensations and energy intake compared to animal protein. Furthermore, not many studies have
used solid and real foods. In a randomized single-blinded cross-over study, no differences in
energy intake were found 30 min after a liquid preload of 20 g casein, whey, pea protein, or egg
albumin [21]. Lang et al. observed no differences in hunger, satiety, ad libitum energy intake, or 24-h
energy intake between lunches enriched with egg white protein (albumin), cow milk protein (casein),
gelatin, soy, wheat or pea protein [22]. The study was, however, limited by different energy content and
macronutrient composition in the test meals. We previously investigated the effect of meals containing
Nutrients 2018, 10, 96 9 of 11

fava beans and split peas as the vegetable protein source compared to pork and veal meat as the animal
protein source [8]. In that study, higher satiety and fullness ratings and lower prospective food intake
and hunger ratings were found after the fava bean and split pea meal compared to the veal and pork
meal. Furthermore, participants had a lower energy intake after the fava bean and split pea meal
compared to the veal and pork meal at the ad libitum lunch served three hours after the test meal.
In that study, dietary fiber content was not matched but reflected the natural content of the included
test foods. Dietary fiber is known to influence appetite [23,24], in this study we matched the fiber
content of the meals. This is probably the main reason for the different outcomes of the two studies.
Few studies have investigated the effect of egg on appetite and energy intake compared to other
protein sources. Our results indicated that egg has similar effects on appetite as meat (pork and veal)
and vegetable proteins (fava beans and split peas). This seems inconsistent with some results from
previous studies [9,10]. Yet, these studies served the meals as liquid breakfasts or liquid preloads.
Liquid meals are known to have less effect on satiety compared to solid meals and this could, therefore,
have influenced the results [25]. Bayham et al. compared an egg breakfast to an energy density and
macronutrient-matched cereal breakfast and observed no difference in energy intake at a subsequent
lunch served three hours after the test meal. However, the participants felt more full 30 min after the
egg breakfast compared to the cereal breakfast [11]. These results are, at least partly, in agreement with
our findings.
Since the greenhouse gas emissions associated with meat production are high compared
to vegetable production, a reduction in consumption of protein from animal sources is highly
recommended [4,5]. In our study, we found a comparable effect on ad libitum energy intake and
subjective appetite sensations after the vegetable protein meal that included fava beans and split peas
and the fiber-supplemented meals with eggs or veal and pork, and the non-fiber-supplemented egg
meal. This demonstrates that legumes are a good alternative to meat with regard to the effect on
appetite. Additionally, intake of legumes may also be beneficial for weight management. A recent
meta-analysis of randomized controlled trials found diets with legumes resulted in better weight
loss (−1.74 kg) and weight maintenance (−0.29 kg) compared to control diets [26]. However,
as demonstrated in our current study, a disadvantage of legume-based meals is the lower palatability.
This could be an obstacle for increasing the intake of vegetable protein in the population, especially in
the Western world where legumes are not part of a traditional diet.
Our findings should be interpreted while considering the following. The test meals did not
represent a typical Danish breakfast, which could have influenced energy intake, subjective appetite
sensations, and palatability ratings. The participants were all males and were constrained to the
laboratory which reduces the generalizability of the results. However, we chose only to include
men to obtain a homogenous group, and for practical reasons, as the time point where women
are tested in their menstrual cycle may affect energy intake (higher energy intake during the luteal
compared to the follicular phase) [27]. A further limitation is the inclusion of both normal weight and
overweight participants.

5. Conclusions
In conclusion, iso-caloric, macronutrient-balanced, fiber-matched meals based on vegetable
protein from fava beans and split peas or animal protein from veal and pork or eggs, were found to
have similar effects on ad libitum energy intake and subjective appetite sensations. Also, a similar
meal based on eggs without supplemented fiber did not differ from the fiber-matched meals.

Supplementary Materials: The following are available online at www.mdpi.com/2072-6643/10/1/96/s1,

Table S1: Mean differences between the four test meals in subjective appetite sensations, well-being, palatability,
and compensatory eating; Figure S1: The four test meals.
Acknowledgments: The study was financed by a grant from the Danish Agriculture and Food Council.
Fibradan® was donated by DLG Food A/S, Copenhagen, Denmark. The authors thank the participants and
Nutrients 2018, 10, 96 10 of 11

the kitchen staff at Department of Nutrition, Exercise and Sports, University of Copenhagen, for creating and
preparing test meals, and Nathalie T. Bendsen for assistance with the design of the study.
Author Contributions: A.A., A.B. and A.R. conceived and designed the experiments; M.D.K. and L.K. performed
the experiments; L.V.N. and C.R. analyzed the data; L.V.N. wrote the paper.
Conflicts of Interest: A.A. is currently consultant/member of advisory boards for Dutch Beer Institute, NL;
Feast Kitchen A/S, DK; Groupe Éthique et Santé, France; McCain Foods Limited, USA; Nestlé Research Center,
Switzerland; Weight Watchers, USA. A.A. has received research grants from Arla Foods, DK; Danish Dairy
Research Council; Gelesis, USA and currently collaborating with Gelesis, USA on commercialization of research
results. He is member of International Carbohydrate Quality Consortium (ICQC), University of Toronto, Canada.
He is co-owner of University of Copenhagen spin-out company Personalized Weight Management Research
Consortium ApS (Gluco-diet.dk) and of patents pending to the University. He receives royalties for several books
in about diet and personalized nutrition for weight loss. None of the other authors declared a conflict of interest.

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