Livestock Science 183 (2016) 2427

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Livestock Science
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Short communication

Effect of Brazilian red pepper (Schinus terebinthifolius Raddi) essential

oil on performance, diarrhea and gut health of weanling pigs
Franz Dias Gois a, Pedro Leon Gomes Cairo a, Vincius de Souza Cantarelli b,
Larissa Corra do Bomm Costa a, Renato Fontana a, Ivan Bezerra Allaman a,
Maicon Sbardella c, Fernando Morais de Carvalho Jnior b, Leandro Batista Costa d,n
a

Universidade Estadual de Santa Cruz, 45662-900 Ilhus, BA, Brazil

Universidade Federal de Lavras, 37200-000 Lavras, MG, Brazil
c
Universidade de So Paulo (USP), Escola Superior de Agricultura Luiz de Queiroz (ESALQ), 13418-900 Piracicaba, SP, Brazil
d
Pontifcia Universidade Catlica do Paran, Escola de Cincias Agrrias e Medicina Veterinria, 83010-500 So Jos dos Pinhais, PR, Brazil
b

art ic l e i nf o

a b s t r a c t

Article history:
Received 2 February 2015
Received in revised form
6 November 2015
Accepted 10 November 2015

This study aimed to evaluate the effects of dietary Brazilian red pepper (Schinus terebinthifolius Raddi)
essential oil and an antimicrobial agent on weanling pig growth performance, diarrhea occurrence, pH of
the digestive content, small intestine histology, and intestine microbial counts. Ninety weanling castrated
male pigs (5.67 0.78 kg BW and 21-d old) were used in a randomized complete block design experiment
with ve treatments, six replications per treatment, and three animals per experimental unit (pen). The
treatments were a basal diet supplemented with 0 (negative control), 500, 1000, and 1500 mg/kg Brazilian red pepper essential oil vs with 120 mg/kg chlorohydroxyquinoline (antibiotic treatment). At the
end of the experimental period one animal from each pen was slaughtered to record the pH of digestive
contents, small intestine histology, and intestine microbial counts. Treatments had no effect (P4 0.05) on
growth performance, diarrhea occurrence, pH of the digestive content, villus height, crypt depth, and
intestinal microbial counts of weanling pigs. However, pigs fed the diet containing 500 mg/kg essential
oil had greater villi density (P o0.05) than those fed diets containing the antibiotic or 1000 and 1500 mg/
kg essential oil. Thus, both Brazilian red pepper essential oil and the antibiotic are of limited benet for
enhancing the growth of weanling pigs. However, the effectiveness of growth enhancer additives may be
reduced in low challenging situations as demonstrated by the current study.
& 2015 Elsevier B.V. All rights reserved.

Keywords:
Additives
Microbiota
Performance
Phytogenics
Piglets

1. Introduction
The use of natural additives for food preservation and disease
control in humans, animals, and plants are of scientic and industrial interest. Continued advances in modern technology are
facilitating the isolation and characterization of active components
present in plants, along with improving our understanding about
their potential actions and, hence, human-interest applications
(Costa et al., 2013).
Traditionally, antibiotic growth promoters were used to enhance piglet performance during weaning period; however,
n

Corresponding author.
E-mail addresses: fdgois16@gmail.com (F.D. Gois),
peucairo@hotmail.com (P.L.G. Cairo), vinicius@dzo.ua.br (V. de Souza Cantarelli),
larissa@uesc.br (L.C. do Bomm Costa), rfontana@uesc.br (R. Fontana),
ivanalaman@gmail.com (I.B. Allaman), msbardella@gmail.com (M. Sbardella),
fernandomvet@yahoo.com.br (F.M. de Carvalho Jnior),
batista.leandro@pucpr.br (L.B. Costa).
http://dx.doi.org/10.1016/j.livsci.2015.11.009
1871-1413/& 2015 Elsevier B.V. All rights reserved.

possible bacterial cross-resistance has limited the use of the antibiotics as growth promoters (Brenes and Roura, 2010). Thus, the
potential application of plant extracts, essential oils, oleoresins,
and their puried compounds is being investigated as alternative
feed strategies to enhance weanling pig performance during this
critical period (Hashemi and Davoodi, 2011).
Brazilian red pepper (Schinus terebinthifolius Raddi, Anacardiaceae) exhibits antimicrobial activity in vitro (Lima et al., 2006) and
also has various medicinal properties, including antioxidant
(Bendaoud et al., 2010), antitumoral (Matsuo et al., 2011) and
antifungal (Johann et al., 2010) properties. However, there is in the
literature, one study with Brazilian red pepper in broiler feed. Silva
et al. (2010) found an increase on weight gain and nal body
weight when broiler chickens were fed dietary red pepper essential oil (4000 mg/kg). Therefore, it has been hypothesized that
red pepper could replace performance-enhancing antibiotics also
in weanling pig diets. To our knowledge there are not studies that
evaluated Brazilian red pepper in swine feed.
This study aimed to evaluate the effects of dietary Brazilian red

F.D. Gois et al. / Livestock Science 183 (2016) 2427

pepper (S. terebinthifolius Raddi) essential oil and an antimicrobial

agent on the growth performance, diarrhea occurrence, pH of digestive content, small intestine histology, and intestine microbial
counts of weanling pigs.

2. Material and methods

2.1. Essential oil composition of Brazilian red pepper (S. terebinthifolius Raddi) fruit
The composition of the essential oil from pepper tree fruit was
determined by using a gas chromatograph (6890N, Agilent Technologies, Santa Clara, USA) with a mass detector (5973, Agilent
Technologies), equipped with a DB-5MS silica column (60 m
0.
32 mm
1.0 m). Identication was conrmed using standard
compounds that were available in the chosen library (Givaudan
Ltda., So Paulo, SP, Brazil). The essential oil from pepper tree fruit
contained -3-carene 41.01%, -phellandrene 14.40%, limonene
12.36%, -pinene 10.36%, and other compounds 21.87%.
2.2. Animals, facilities, and experimental design
All procedures using animals were approved by the Committee of Ethics for the Use of Animals (CEUA) of the State University
of Santa Cruz (Ilhus, Bahia, Brazil) (Protocol 024/2011). Ninety
weanling castrated male pigs (female DanBred DB90
male PICAGPIC337) (5.67 0.78 kg BW and 21-d of age) were used in a
randomized complete block design experiment with ve treatments, six replications per treatment, and three animals per experimental unit (pen).

25

Table 1
Basal diet composition (as-fed basis) fed to piglets throughout the 35-day experimental perioda.
Ingredients (%)

Pre-starter diet
(114 days)

Starter diet
(1435 days)

Soybean meal (46%)

Corn
Gelatinized corn
Biscuit meal
Milk productb
Milk productc
Dextrin
Sugar
Soybean oil
Dicalcium phosphate
Limestone
Salt
NaHCO3
L-lysine.HCl (78%)
DL-Methionine (99%)
L-Threonine (98.5%)
L-Tryptophan (98%)
Choline chloride (60%)
Vitamin and mineral premixd
Inert and/or feed additive
Calculated values (%)
Metabolizable energy (MJ/kg)
Crude protein
Calcium
Total phosphorus
Available phosphorus
Digestible lysine
Digestible met cys
Digestible tryptophan
Digestible threonine

31.02
16.11
15.00
20.00
5.00
3.16
4.10
0.21

1.88

0.40
0.25
0.55
0.31
0.20
0.02
0.10
0.20
1.50

33.71
43.63

6.00
4.00
1.50
2.20
2.60
1.27
1.59
0.42
0.40
0.20
0.40
0.23
0.11

0.04
0.20
1.50

14.57
20.23
0.85
0.71
0.51
1.46
0.83
0.88
0.25

14.15
20.63
0.83
0.66
0.45
1.37
0.79
0.82
0.24

a
b

2.3. Experimental diets

Commercial diet-Master Nutrio Animals (Minas Gerais, Brazil).

Start-pro 20 (40.5% lactose)-Auster Nutrio Animal Ltda. (So Paulo, Bra-

zil).
c

The dietary treatments were a basal diet supplemented with 0

(negative control), 500, 1000, and 1500 mg/kg of red pepper essential oil (AgroRosa Ltda, So Mateus, ES, Brazil) vs 120 mg/kg of
chlorohydroxyquinoline (antibiotic treatment) (Indukern Ltda, So
Paulo, SP, Brazil). The red pepper essential oil was microencapsulated to stabilize the molecule and minimize its sharp
avor. The microcapsules contained 15% of the red pepper essential oil and the carrier.
Basal diets (Table 1) were formulated according to the nutrient
requirements of pigs (Rostagno et al., 2011). A two-phase feeding
program was adopted: pre-starter (days 114 of the experiment)
and starter (days 1435 of the experiment).

Prius L72 (71.5% lactose)-Auster Nutrio Animal Ltda. (So Paulo, Brazil).
Quantities per kg of feed: 50 mg of manganese; 160 mg of zinc; 246 mg of
iron; 14 mg of copper; 1.5 mg of iodine; 15000 IU of vitamin A; 3000 IU of vitamin
D3; 110 IU of vitamin E; 5.5 mg of vitamin K3; 4.4 mg thiamine; 9.2 mg of riboavin; 6.6 mg Pyridoxine; 38 g of vitamin B12; 1.6 mg of folic acid; 27 mg of
Pantothenic acid; 44 mg of niacin; 0.14 mg of biotin; 0.36 mg of selenium.
d

Table 2
Effects of dietary Brazilian red pepper essential oil and an antimicrobial agent on
the body weight (BW), average daily feed intake (ADFI), average daily gain (ADG),
feed:gain ration (F:G), diarrhea occurrence (DF) (n 18/group), and villi density
(VD) in the duodenum and jejunum (n 6/group) of weanling piglets during the
35-day post-weaning period.
Variables

2.4. Experimental procedure

During the trial period, the animals were provided water and
feed ad libitum.
The performance variables (average daily feed intake, average
daily gain, and feed:gain ratio) were calculated by using animal
BW and feed disappearance data. The presence or absence of
diarrhea (liquid feces on the oor and/or soiling in the anal area)
was checked twice a day to calculate the percentage of days with
diarrhea.
At the end of the experiment one animal with BW closest to the
average BW of each pen (with six animals per treatment) was
slaughtered according to humanitarian approved methods by
electrical stunning followed by exsanguination. The pH of the
stomach, jejunum, and cecum contents was immediately measured by inserting a unipolar electrode, following the methods of
Manzanilla et al. (2004).
To measure the histology of the small intestine (villus height
and crypt depths), 3-cm samples of duodenum (resected at 15 cm

Performance:
Initial BW (kg)
Final BW (kg)
ADFI (g)
ADG (g)
F:G
DF (%)
VD duodenumc
VD jejunum

Treatmentsa
ANT

500

1000

1500

5.64
14.85
497.82
321.23
1.71
36.00
25.44c
32.75

5.65
16.99
557.29
311.56
1.80
37.50
33.75ab
32.22

5.66
16.30
543.14
345.53
1.72
37.83
39.56a
28.89

5.65
14.84
478.88
308.65
1.61
38.17
18.29c
27.00

5.65
16.66
538.84
323.79
1.90
28.00
26.56bc
37.50

SEMb

P-value

0.078
103.9
34.88
0.001
2.692
0.81
2.55

0.10
0.14
0.36
0.16
0.25
0.00
0.31

a
ANT Antibiotic (120 mg/kg chlorohydroxyquinoline); 0; 500; 1000; or
1500 mg/kg of red pepper essential oil.
b
Standard Error of the Mean.
c
Different letters in the row indicate a signicant difference between the values by the Tukey test (5%).

from the pylorus sphincter) and jejunum (resected at 150 cm from

the ileocecal junction) were cut, washed with saline solution (0.9%
NaCl), and xed in buffered formalin solution as described by Gao

26

F.D. Gois et al. / Livestock Science 183 (2016) 2427

Fig. 1. Electron-micrographs of the duodenum mucosa of piglets at 35 days of age. (ANT Antibiotic; 0; 500; 1000; or 1500 mg/kg of Brazilian red pepper essential oil)

et al. (2000). To analyze the intestine ultrastructure, the duodenum and jejunum samples were processed following the methodology described by Rigueira et al. (2013). The best images from
each sample were chosen for the measurement of villi density
(number of villi per area) (villi/847.882 mm2).
For the microbial counts, samples from the jejunum (150 cm
from the ileocecal junction) and cecum (homogenized content)
were placed in sterile containers by using a glass slide scraping.
Total counts of aerobic bacteria, Enterobacteria and Lactobacillus,
were determined within 24 h as described by Montagne et al.
(2012).
2.5. Statistical analysis
The data were submitted to analysis of variance, considering a
randomized complete block design experiment and the pen as the
experimental unit. When P o0.05, the means were compared by
Tukey. R software (R Development Core Team, 2014) was used for
statistical analysis.

3. Results and discussion

3.1. Growth performance, diarrhea occurrence, pH of the digestive
contents and microorganism counts
Growth performance, diarrhea occurrence (Table 2), pH of the
contents of the stomach, jejunum, and cecum and microorganism
counts (log UFC/g) of the content of jejunum and cecum were not
inuenced (P 40.05) by the dietary treatments.
Data from this study corroborate those obtained by previous
studies. For instance, the positive effects of plant-based additives
on performance were not detected following the application of
dietary essential oil extracted from Origanum vulgare (Henn et al.,
2010), Foeniculi aetheroleum, or Carvi aetheroleum (Schne et al.,
2006). Santana et al. (2015) also found no improvement in the
performance of piglets fed with 40 mg/kg of colistine sulfate. In

other study, Perina et al. (2014) observed that the antimicrobial

treatment (120 ppm of chlorhydroxyquinoline), the same used in
the current experiment, reduced BW and G:F of weanling pigs
from 13 to 27, 27 to 40, and 1 to 40 d, compared to control
treatment. The lack of positive results of chlorhydroxyquinoline
may be due to rigorous sanitary control of the facilities, low
challenging situations. Besides, the possible broad-spectrum activity of this antimicrobial may have caused the imbalance between benecial and pathogenic microbiota and, consequently,
show no benecial effects.
There are no records in the literature regarding the use of red
pepper in swine feed. However, broiler chickens fed dietary red
pepper essential oil (4000 mg/kg) exhibited increased weight gain
and nal body weight (Silva et al., 2010). We supplemented it in
low doses, since that the properties of this essential oil can reduce
of palatability of feed, changing the feed avors and causing feed
aversion in swines. Although the major compounds present in the
red pepper essential oil in this preceding study were similar to
those in the current study, there was wide variation in the concentration of these compounds; -pinene (29.39% vs 10.36% in the
previous and current study, respectively), -3-carene (19.69% vs
41.01%), limonene (18.15% vs 12.36%), and -phellandrene (9.39%
vs 14.40%). This variation might explain the differences in the
growth-enhancing effects observed between these two studies,
along with the fact that different animal species, experimental
conditions and level (0, 500, 1000 and 1500 mg/kg) of red pepper
essential oil used, the plant parts used for essential oil extraction
(Ao et al., 2011), the extraction method (Barbosa et al., 2007) and
variations in plant parameters. No effect of dietary Brazilian red
pepper essential oil or chlorohydroxyquinoline on the occurrence
of diarrhea was observed, according to Manzanilla et al. (2004)
and Henn et al. (2010).
Plant-based feed additives showed no signicant effect on the
pH of stomach (3.127 0.84), jejunum (6.20 70.08) and cecum
(6.17 0.14) content of weanling pigs, supporting previous study
(Costa et al., 2011). In this current study, the addition of the mineral and protein sources to the diet might have enhanced the

F.D. Gois et al. / Livestock Science 183 (2016) 2427

buffering capability, preventing any pH variation in the digestive

content.
Piglets fed dietary antibiotics and Brazilian red pepper essential
oil had not reduced total aerobic bacteria (18.0 72.92), enterobacteria (18.0 72,26) and lactobacillus (18.0 75,21) (log cfu/g)
in the jejunum, and total aerobic bacteria (11.15 70.08), enterobacteria (10.68 70.43) and lactobacillus (11.04 70.09) (log cfu/
g) in the cecum of weanling pigs. Data from this study corroborate
those obtained by Li et al. (2012) that used essential oil in piglets
diets evaluating total aerobic bacteria, enterobacteria and lactobacillus counts (log cfu/g).
3.2. Small intestine histology
Piglets fed dietary 500 mg/kg red pepper essential oil had
higher villi density (number of villi per specic area)
(villi/847.882 mm2) in the duodenum (P o0.05) than those fed
dietary 1000 and 1500 mg/kg essential oil or the antibiotic (Table 2) (Fig. 1). Greater densities of villi enhance the ability to digest
and absorb nutrients. This phenomenon is explained by the presence of more enterocytes, which increase the absorptive surface.
No effects of dietary Brazilian red pepper essential oil or
chlorohydroxyquinoline were observed on villi height in duodenum (229.56 710.78 mm) and jejunum (244.507 17.12 mm), crypt
depth
in
duodenum
(101.26 75.17 mm)
and
jejunum
(112.13 78.41 mm), and villi height:crypt depth in duodenum
(2.32 70.09 mm) and jejunum (2.28 7 0.10 mm), supporting previous reports (Li et al., 2012) that used plant-based additives in
weanling pig diets.

4. Conclusions
The results of this study revealed that neither Brazilian red
pepper essential oil nor the antibiotic were of any benet to the
performance of weanling pigs. However, the effectiveness of
growth enhancer additives may be limited in low challenging situations or rigorous sanitary control of the facilities, as observed in
the current study. Therefore, further information is required on the
compounds present in Brazilian red pepper essential oil, in addition to their effects and potential application under different
conditions in swine feed.

Conict of interest
All authors declare that there are no conicts of interest concerning to the information provided in this paper.

Acknowledgments
We acknowledge the Fundao de Amparo Pesquisa do Estado da Bahia-FAPESB for granting aid for this research-APP0052/
2011.

27

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