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German Journal of Veterinary Research
eISSN:2703-1322
Research article
Effectiveness of standardized red orange extract (Citrus sinensis) for weight
reduction in canine obesity
Emerson Milla∗and Ivo I. Kerppers
Postgraduate Studies Program in Veterinary Sciences, Department of Veterinary Medicine, Universidade Estadual do Centro
Oeste, Guarapuava, Paran´a, Brazil
Article History:
Received: 01-Aug-2023
Accepted: 28-Aug-2023
*Corresponding author:
Emerson Milla
emerson.milla@uniguairaca.edu.br
Abstract
Obesity is the most common nutritional disease in both humans and dogs, currently
characterized as a low-intensity inflammatory state. The anthocyanins found in red
oranges can assist in combating weight gain and reducing body fat accumulation by
reducing lipogenesis and modulating inflammation. This study aimed to compare the
effectiveness of standardized dried red orange extract in reducing overweight in obese
dogs compared to chromium picolinate and a placebo. In total, 23 animals were used
in the current study and divided randomly into three experimental groups. All dogs
received the same hypocaloric diet and supplementation: standardized Citrus sinensis
red-orange extract (G1, n=9), chromium picolinate (G2, n=7), and a placebo-control
group (G3, n=7). The dogs were managed by their owners for 90 days. In the
distribution of the body condition score (BCS) determined at the beginning of the
study, 47.8% of the animals had BCS of 6; 26.1% had BCS of 7; 8.7% had BCS of
8, and 17.4% had BCS of 9. During the program, most of the owners reported being
unable to involve other residents of their households in the dogs’ treatment. At the
end of the experiment, the G1 group had a significant reduction in weight and BCS
(p<0.01), as well as in HDL cholesterol levels (p<0.05) when compared to the other
groups. It was concluded that incorporating dried red-orange extract into the diet of
overweight and obese dogs can lead to weight loss and improved BCS.
Keywords: Companion animals, Citrus sinensis, Dogs, Nutraceuticals, Obesity, Red orange
extract
Citation: Milla, E. and Kerpper, I. I. Effectiveness of standardized red orange extract
(Citrus sinensis) for weight reduction in canine obesity. Ger. J. Vet. Res. 3 (2):35-40.
https://doi.org/10.51585/gjvr.2023.2.0057
Introduction
Obesity is currently the most common nutritional dis-
ease in companion animals, and in dogs, it occurs when
the body weight is at least 15% above the ideal weight
(Burkholder and Toll,2000). In both humans and com-
panion animals, the medical significance of obesity lies
in its effect on the mortality and morbidity of associ-
ated diseases (Suarez et al.,2022). Overfeeding is the
main reason for developing the disease in humans and
dogs, which can lead to a positive energy balance, re-
sulting in excess body fat storage (Carciofi et al.,2005).
In its current concept, obesity has been seen as a
low-intensity inflammatory state directly or indirectly
associated with cardiovascular, respiratory, and ortho-
pedic problems and metabolic disorders such as re-
duced glucose tolerance, diabetes mellitus, and dys-
lipidemia (German,2010).
Natural agents, like red-orange Citrus sinensis (L.)
Osbeck has been researched as an alternative for re-
ducing excess adipose tissue. It is rich in phenolic
compounds, vitamin C, and anthocyanins, which act as
powerful antioxidants, modulating inflammation gener-
ated by excess adipose tissue and inhibiting lipid per-
oxidation (Rodrigues et al.,2020;Briskey et al.,2022).
Recent studies have shown that the standardized red-
orange extract can downregulate adipogenic genes and
enzymes, along with modulating adiponectin secretion
and leptin release. Consumption of red oranges (espe-
cially Moro juice) has also been found to curb body
weight gain, improve insulin sensitivity, and reduce
serum levels of triglycerides and total cholesterol in
mice (Titta et al.,2010;Salamone et al.,2012).
While severe overweight conditions in companion
animals are readily noticeable, detecting subtle changes
presents a challenge for many methods. This makes
monitoring weight loss programs and early interven-
tions complex. The assessment of body condition aims
to determine whether animals are underfed or overfed.
35
Table 1: Body condition scores in dogs (Laflamme,2006).
Body condition Scores Description
Underfed
Ribs, lumbar vertebrae, pelvic bones, and all bony prominences are visible from a dis-
tance. There is no discernible body fat. Evident loss of muscle mass.
Ribs, lumbar vertebrae, and pelvic bones are easily visible. No palpable fat. Some other
bony prominences may be visible. Minimal loss of muscle mass.
Ribs are easily palpable and may be visible without palpable fat. Top of lumbar ver-
tebrae visible. Pelvic bones start to become visible. Waist and abdominal tuck are
evident.
Ideal Ribs are easily palpable with minimal fat covering. From a top view, the waist is easily
observed. Abdominal tuck evident.
Palpable ribs without excess fat covering. Abdomen tucked when viewed from the side.
Overfed
Palpable ribs with slight excess fat covering. The waistline is visible when viewed from
the top but not pronounced. Abdominal tuck is apparent.
Palpable ribs with difficulty; intense fat covering. Evident fat deposits over the lumbar
area and base of the tail. Absence of waist or only slightly visible. Abdominal tuck may
be present.
Inability to palpate the ribs located under a very dense fat cover or ribs palpable only
with firm pressure. Heavy fat deposits over the lumbar area and base of the tail. No
waistline. No abdominal tuck. Significant abdominal distension may be present.
Massive fat deposits over the chest, spine, and base of the tail. Fat deposits on the neck
and limbs. Abdominal distension is evident.
Numerous techniques are available for evaluating body
composition, including advanced methods like dual-
energy X-ray absorptiometry (DEXA), magnetic res-
onance, neutron activation analysis, and computed to-
mography. However, only a few of these options are
appropriate for animal use (Diez and Nguyen,2006;
Andrade Junior et al.,2020).
Using body weight as an indicator of body compo-
sition is limited by significant variation among breeds,
age ranges, and sex, and it is not recommended as the
sole form of evaluation as it does not assess fat mass
or muscle mass. Dogs with the same body weight can
have different body compositions. Body weight can
only quantify the percentage of weight excess by com-
paring it to the ideal weight, which is often a theoretical
value (Diez and Nguyen,2006). Body condition score
(BCS) is used to subjectively and semi-quantitatively
assess an animal’s body fat and muscle mass. The BCS
(Table 1) uses numerical scales based on patient inspec-
tion and palpation, employing scales from one to nine
to reduce subjectivity (Laflamme,2006).
The long-term weight control of obese pets can rep-
resent a frustrating clinical challenge destined to fail
unless an organized approach with practical strategies
is adopted (Center,2003;Rodrigues et al.,2020).
There needs to be more research focusing on ex-
ploring novel weight loss solutions for dogs beyond the
commonly recommended methods of a well-rounded
diet and regular physical activity. This study aimed to
evaluate the weight-reducing properties of red-orange
extract, compared to chromium picolinate and a bal-
anced diet alone, for animals.
Materials and methods
Ethical approval
This study was approved by the Ethics Committee
on Human Research, COMEP/UNICENTRO (proto-
col 5.180.123), and by the Committee on Ethics in
the Use of Animals (CEUA) with protocol number
01/2022.
A questionnaire was created using the Google
Forms platform, consisting of 16 objective multiple-
choice questions. It was designed to be easy to un-
derstand and was administered to overweight or obese
dog owners at two veterinary clinics in the city of
Guarapuava-PR.The owners were informed about the
research and voluntarily responded to the question-
naire. The questionnaire was provided from January
21st to March 15th, 2022. Forty-five owners responded
to the questionnaire.
Selection of the animals
The inclusion criteria for participation in the project
were: dogs aged between 2 and 7 years, healthy, with-
out comorbidities with a body condition score (BCS)
between 6 and 9, of both sexes, spayed or not, and
weighing between 5 and 40 kg. The owners were asked
about the body condition of their dogs, and those who
agreed to participate in the project (n=23) signed an
informed consent form.
Exclusion criteria encompassed dogs with a his-
tory of significant medical issues, including cardiovas-
cular, neurological, renal, gastrointestinal, immune,
endocrine, or hematological abnormalities, and those
with a BCS≤5. Animals receiving systemic or topical
glucocorticoids and anticonvulsants were also excluded.
36
Table 2: Data from experimental groups (mean±standard deviation)
Group Number Mean age Sex (%) Body condition
(years) Males Females scores a
Morosil®9 5.89±1.96 22.2 77.8 7.0±1.39
Chromium picolinate 7 4.43±2.07 14.3 *71.4 7.0±0.9
14.3 *
Placebo 7 4.21±1.58 28.6*42.9 6.0±1.11
28.6*
Total 23 4.93±1.97 8.7 65.2 6.96±1.15
13.0*13.0*
aBody condition scores after Laflamme (2006). *Indicates the percentage of non-castrated animals
Table 3: Difficulties reported by owners during the program.
Question Group
Morosil®Chromium picolinate Placebo Total
Are owners following Response Yes No Yes No Yes No Yes No
the program correctly? Frequency 6 3 1 6 4 3 11 12
% 66.7 33.3 16.7 83.3 57.2 42.8 47.8 52.2
Did owners manage to involve other Frequency 4 5 3 4 2 5 9 14
household members in treating their dogs? % 44.5 55.5 42.8 57.2 28.5 71.5 39.1 60.9
Do owners consider that their Frequency 2 7 1 6 3 4 7 16
dogs have been hungry? % 22.2 77.8 14.2 85.8 42.8 57.2 30.4 69.6
On day 60 of the study, a second questionnaire con-
sisting of three multiple-choice questions was adminis-
tered to the owners. This secondary assessment aimed
to gain insight into the challenges they encountered
during their animals’ weight loss program.
At the program’s commencement and conclusion,
the dogs were weighed, and anamnesis data and BCS
were documented on an evaluation form. Blood sam-
ples were collected at the program’s onset and conclu-
sion through punctures to the cephalic or jugular vein,
with 5 ml of blood drawn and placed in anticoagulant-
free tubes. The samples were appropriately labeled
and refrigerated before transport to the Animal Lab
Laboratory in Guarapuava-PR.
Serum biochemical parameters assessed included
total cholesterol, its fractions (HDL, LDL, and VLDL),
and triglycerides. An automated biochemical analyzer
(IDEXX Catalyst One ®, Maine, USA) was used for
analysis.
Experimental groups
The animals were randomly divided into three groups.
Group 1 (G1, n=9) received a specific and individu-
alized diet for each dog, along with dry red orange
extract (Morosil®) at a dose of 200 mg/animal/day
orally (PO), as directed by the supplier (Galena,2021).
The standardized extract was provided by Bionap S. R.
L. (Belpasso, Catania, Italy).
Group 2 (G2, n=7) received a specific and indi-
vidualized diet for each dog, along with chromium pi-
colinate (Sigma-Aldrich, Saint Louis, USA) at a dose
of 0.01 mg/kg/day PO (NRC,2006). Group 3 (G3,
n=7) was assigned an individualized diet for each dog,
supplemented with maltodextrin (Sigma-Aldrich, Saint
Louis, USA) at a dosage of 200 mg/animal/day orally,
intended as a placebo. At this dose, maltodextrin holds
no nutritional value and does not induce changes in
blood biochemical parameters. Thus, G3 functioned
as the control group.
The Morosil®, chromium picolinate, and placebo
were manipulated and distributed to the participants
in similar sachets, identical in appearance and function,
and without identification, with the instruction to be
given to the dogs together with their first meal of the
day. In all three cases, a salmon-flavored palatabilizer
was used.
Energy requirements and animal monitoring
The feed used in this experiment contained 10% mois-
ture, 35.5% crude protein, 8% ether extract, 7.5%
ash, 15% crude fiber, 1.3% calcium, 0.6% phosphorus,
0.66% potassium, 1.5% omega-6, and 0.3% omega-3.
The feed was provided to the owners free of charge,
and each owner received a measuring cup adjusted for
their dog to facilitate the proper daily feeding amount,
divided into one, two, or three portions, according to
their convenience. The amount of feed offered to the
animals corresponded to 70% of their maintenance en-
ergy requirement (NRC,2006), calculated for their tar-
get weight, standardized as their current weight minus
10%.
The experimental groups were managed by the
owners themselves, who received instructions to follow
the weight loss program for 90 days. It was also rec-
ommended that the dogs not be given treats and other
foods during the experimental period. The follow-up
was done through monthly weighting of the animals,
interviews with the owners, and periodic contact via
telephone to encourage them to continue with the pro-
gram.
37
Table 4: The initial and final average weight (mean±standard deviation) evaluated by repeated measures
ANOVA at a significance level of 5% post-Tukey test.
Group Number Average initial weight Average final weight F*p-value
Morosil®9 20.7±12.6 19.5±12.3 9.42 <0.01
Chromium picolinate 7 16.4±10.1 15.2±8.95 4.71 0.013
Placebo 7 11.1±5.48 10.9±5.64 0.308 0.820
*F; value obtained from the repeated measures analysis of variance..
Table 5: The initial and final body condition score (BCS) expressed as mean±standard deviation assessed
through the Friedman test and multiple Durbin-Conover comparisons with 5% significance.
Group Number Initial BCS Final BCS p-value
Morosil®9 7±1.39 6±1.27 <0.001
Chromium picolinate 7 7±0.9 6±0.976 0.078
Placebo 7 6±1.11 6±0.690 0.078
Statistical analysis
The interview data were tabulated in Microsoft Excel®
spreadsheets, and the results were expressed by de-
scriptive statistics, presenting the values in frequency.
A repeated-measures ANOVA test was used to evalu-
ate the variation in the animals’ weight. Significance
was considered at 5% post-Tukey’s test. The Friedman
test was used to evaluate the animals’ body condition
score. Significance was considered at 5% post multiple
comparisons of Durbin-Conover. For the blood bio-
chemistry tests, the Wilcoxon test was used with a sig-
nificance level of 5%. The statistical analysis was per-
formed using the Jamovi software (Anonymus,2020).
Results
The data from the studied dogs are presented in Ta-
ble 2. The average age of the study population (n=23)
was 4.93±1.97 years old. In the group that received the
dry extract of red orange (n=9), the average age was
5.89±1.96 years. In the group that received chromium
picolinate (n=7), the average age was 4.43±2.07 years,
and in the placebo group (n=7), the average age was
4.21±1.58 years.
The BCS of the animals was assessed at the initial
consultation, and 47.8% of the animals were found to
have a BCS of 6; 26.1% had a BCS of 7; 8.7% had a
BCS of 8 and 17.4% had a BCS of 9. In the evalua-
tion of the BCS of the animals in the Morosil®group,
an average BCS of 7±1.39 was found. Analyzing the
BCS frequencies revealed that 44% of the animals had
a BCS of 6; 22.2% had a BCS of 7, and 33.3% had a
BCS of 9.
In the group that received chromium picolinate, the
average BCS was 7±0.9, with a distribution of 42.9%
of animals with a BCS of 6; 28.6% with a BCS of 7, and
28.6% with a BCS of 8. In the control group, which
received a placebo, the average BCS was 6.96±1.15,
with 57.1% of animals having a BCS of 6; 28.6% with
a BCS of 7, and 14.3% with a BCS of 9.
At the 60-day mark of the program, a second in-
terview was conducted with the participating owners,
in which they were asked about the main difficulties
encountered up to that point (Table 3). Less than half
of the participants (47.8%) stated that they followed
the program correctly. One of the major difficulties re-
ported was the inability to involve all household mem-
bers in treating their animals (60.9%). Additionally,
30.4% of the interviewed guardians believed that their
dogs were feeling hungry.
Comparing the initial and final weighings of
the animals (Table 4), the group that received
Morosil®(n=9) showed a statistically significant
weight loss (p<0.01) compared to the group that re-
ceived chromium picolinate (n=7; p=0.013) and the
placebo-controlled group (n=7; p=0.82).
In the comparison of the BCS among the groups
(Table 5), a statistically significant difference was ob-
served in the reduction of BCS between the initial
and final measurements in the group that received red-
orange extract (n=9; p<0.001) when compared to the
chromium picolinate group (n=7; p=0.078) and the
placebo group (n=7; p=0.078).
In the comparison of biochemical parameters
among the three treatments (Table 6), there was no
statistically significant difference, except for the HDL
cholesterol of the group that received Morosil®(n=9;
p<0.05).
Discussion
Based on the results obtained, most of the dogs were
adults, as evidenced in other epidemiological studies
(Carciofi et al.,2005;Courcier et al.,2010;Aptekmann
et al.,2014;Teixeira et al.,2020). As the animal ages,
lean mass is replaced with fat mass, and it is advised
to reduce energy intake by 10 to 15% from the age
of seven, according to the animal’s body condition, as
suggested by Aptekmann et al. (2014). Castration has
been cited as a risk factor for the development of obe-
sity in previous studies, and the hypothesis for this
increased risk is due to a decrease in basal metabolic
rate, changes in eating behavior, and reduced physi-
cal activity (Courcier et al.,2010;Aptekmann et al.,
2014).
A previous study suggested that dogs living in
households with more members were more prone to
developing obesity because there would always be a
temptation, especially for children to offer treats or
leftovers, leading to energy intake above what is desir-
able(Bland et al.,2009). Rohlf et al. (2010) emphasize
38
Table 6: Biochemical parameters (mean±standard deviation) by group.
ParametersaMorosil®Chromium picolinate Placebo
Initial Final Initial Final Initial Final
Cholesterol 168±94.7 131±72.0 159±28.6 132±38.7 146±37.1 128±31.9
HDL 126.6±58.0 101.3±43.3b86.3±28.3 81.6±27.5 101.1±29.4 80.7±18.2
LDL 48.8±35.8 35.3±28.4 58.9±38.1 35.9±21.0 35.9±10.6 36.4±14.3
VLDL 19.5±21.4 16.4±10.25 14.3±10.8 14.4±8.34 8.86±3.21 11.14±7.19
Triglycerides 97.6±106.5 100.8±95.5 70.1±54.9 92.4±65.3 42.1±18.3 55.9±38.2
aReference values (mg/dL): Cholesterol (108-270), HDL Cholesterol (40-78), LDL Cholesterol (31-71), VLDL Cholesterol (<25), Triglyc-
erides (20-150) (Vaden,2009). bp-value <0.05 for the Wilcoxon test.
the need for counseling of the tutors, encouraging the
whole family to be involved in the dog’s weight loss.
Simply pointing out that the dog is overweight and
that something needs to be done about it is unlikely
to lead to satisfactory results. For many families, the
act of feeding and providing treats to dogs is a way of
showing affection. Many owners believe that food re-
striction causes suffering to their dogs, and they prefer
to see their animals ”happy”, even though they know
that excess weight reduces life expectancy. This would
be a way to justify their animals’ body condition and
rationalize the lack of interest in solving the problem
(Larsen and Villaverde,2016).
In the study conducted by Carciofi et al. (2005),
50% of the interviewed owners believed that their dogs
felt hungry during the weight loss program. The behav-
ior of ”begging for food” can be interpreted as hunger,
increasing the chances of providing treats and leftovers,
or even increasing the amount of food.
Mawby et al. (2004) demonstrated the reliability
of BCS when comparing four different body condition
assessment methods using DEXA, deuterium dilution,
BCS, and biometry in 23 healthy dogs. There was a
good correlation (r2=0.92) between the percentage of
fat obtained by DEXA and BCS. DEXA is a technique
used to measure body fat and lean tissue by using two
different energy X-rays (70 and 140 kVp) that enable
differentiation of the type and amount of tissue, as well
as bone mineral density, fat mass, and lean mass of the
body. The low coefficient of variation indicates that it
is a very precise, fast, and safe technique, but due to
the high equipment cost, it is only used experimentally
in research with small animals (Shoveller et al.,2014).
No studies were found that evaluated the effect of
red orange extract on weight control in obese dogs.
Similar results were found in research with humans,
such as in Cardile et al. (2015), in which 60 adults were
supplemented with 400 mg of Morosil®or placebo for
12 weeks. At the end of the experiment, a signifi-
cant reduction (p<0.05) in body weight, body mass
index, and abdominal circumference of the adults
supplemented with Morosil®was reported. Briskey
et al. (2022) evaluated 98 men and women receiving
Morosil®or a placebo for six months in a randomized,
double-blind, placebo-controlled study. Following the
final assessment with DEXA, the authors noted a sub-
stantial decrease in fat mass and abdominal fat com-
pared to the placebo group. The authors attributed
the phyto-complex within Moro orange, primarily com-
posed of flavonoids like anthocyanins, naringenin, and
hesperidin, with the ability to diminish adipocyte lipid
accumulation and modulate cytokine release, thereby
alleviating oxidative stress and inflammation.
In a study that evaluated the effect of Moro or-
ange juice supplementation in rats fed a high-fat
diet for 12 weeks, the results showed a decrease in
weight gain, triglycerides, and total cholesterol, as
well as an improvement in hepatic steatosis (Salam-
one et al.,2012). In another study, Jeusette et al.
(2010) found a significant difference in plasma con-
centrations of cholesterol and triglycerides in cats sup-
plemented with flavonoids (naringenin) compared to
the non-supplemented group. According to the re-
searchers, flavonoids help limit weight gain, decrease
triglyceride synthesis and accumulation in the liver,
lower total cholesterol, VLDL, and glucose concentra-
tions, and improve insulin sensitivity. However, in the
study by Briskey et al. (2022), no difference was found
in the biochemical parameters of 98 human patients
supplemented with Morosil®over six months.
Conclusion
In the present study, the use of red orange extract
(Morosil®) resulted in a reduction of weight and BCS
in obese dogs. The extract is thought to reduce inflam-
mation and oxidative stress associated with adipocyte
hypertrophy. However, additional research is neces-
sary to determine if the supplementation can be rec-
ommended as a truly effective method for treating and
preventing obesity in dogs.
Article Information
Funding. This work was partially funded by AGalena and
Bionap S. R. L., however, they had no role in interpreting data,
concluding findings, or deciding to publish the article.
Conflict of Interest. Emerson Milla is a Master’s degree student
at the Postgraduate Studies Program in Veterinary Sciences,
Department of Veterinary Medicine, Universidade Estadual do
Centro Oeste, Guarapuava, Paran´a, Brazil. The authors declare
no conflict of interest.
Acknowledgment. The authors would like to express their grati-
tude to Universidade Estadual do Centro Oeste (UNICENTRO)
and Galena and Bionap S. R. L. for for the support provided
during this research.
Authors Contributions. Conceptualization EM and IK; method-
ology, EM and IK; formal analysis, EM and IK; investigation,
EM; resources, EM; data curation, EM; writing the original
draft preparation, EM; writing, review and editing, EM and
39
IK; visualization, EM and IK; supervision, EM and IK; project
administration, EM; funding, EM.
Publisher’s Note. The claims and data contained in this
manuscript are solely those of the author(s) and do not rep-
resent those of the GMPC publisher, editors, or reviewers.
GMPC publisher and the editors disclaim the responsibility for
any injury to people or property resulting from the contents of
this article.
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