Dried bovine placenta improves spermatozoa count in a rat model of male reproductive aging
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Veterinary World, EISSN: 2231-0916 RESEARCH ARTICLE
Available at www.veterinaryworld.org/Vol.14/June-2021/25.pdf Open Access
Dried bovine placenta improves spermatozoa count in a rat model of
male reproductive aging
Surya Agus Prihatno1 , Yosua Kristian Adi1 , Teguh Budipitojo2 , Topas Wicaksono Priyo Jr1 and
Yonathan Alvin Maruli Asi Sihotang3
1. Department of Reproduction and Obstetric, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta
55281, Indonesia; 2. Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta
55281, Indonesia; 3. Sains Veteriner Magister, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta
55281, Indonesia.
Corresponding author: Surya Agus Prihatno, e-mail: prihatno@ugm.ac.id
Co-authors: YKA: yosua.kristian.a@ugm.ac.id, TB: budipitojo@ugm.ac.id, TWPJ: topas.wicaksono@ugm.ac.id,
YAMAS: yonathan.alvin.m@mail.ugm.ac.id
Received: 11-01-2021, Accepted: 06-05-2021, Published online: 21-06-2021
doi: www.doi.org/10.14202/vetworld.2021.1602-1607 How to cite this article: Prihatno SA, Adi YK, Budipitojo T,
Priyo Jr TW, Sihotang YAMA (2021) Dried bovine placenta improves spermatozoa count in a rat model of male reproductive
aging, Veterinary World, 14(6): 1602-1607.
Abstract
Background and Aim: In the male reproductive system, the aging process can lead to infertility. Recently, placenta and its
derivatives have been researched as regenerative agents. This study aimed to describe the basic components of dried bovine
placenta powder and its potential effects as a regenerative agent in a rat model of male reproductive aging with D-galactose
induction.
Materials and Methods: We divided 15 male Wistar rats, 2 months of age, into three groups: A, the health control group; B,
the D-galactose induction group, and C, the D-galactose induction and 10% dried bovine placenta supplementation group.
We measured epididymal sperm concentration and testicular weight and volume and analyzed these using one-way analysis
of variance.
Results: Dried bovine placenta was rich in nutrients, with 61.98% protein, 21.25±2.07 carbohydrates, 8.58% water, 4.93%
ash, and 3.27% fat. The mean epididymal spermatozoa concentration of the rats in Groups A, B, and C was 3026×106/mL,
1492.8×106/mL, and 2732.5×106/mL, respectively. The average total testicle weights were 2.44 g, 2.72 g, and 2.57 g,
respectively. The average total testicle volumes were 2.29 cm3, 2.49 cm3, and 2.33cm3, respectively.
Conclusion: Dried bovine placenta powder is rich in nutrients, especially protein. Supplementation with dried bovine
placenta can improve epididymal spermatozoa concentration that is important in fertility.
Keywords: aging, bovine placenta, male reproductive, sperm count, testicular.
Introduction Recently, placenta and its derivatives have been
Aging is a complex biological process involving studied as regenerative therapy for various degenera-
organic, cellular, and molecular changes. This pro- tive diseases. Placental components can include cord
cess is caused chronologically by increasing age but blood cells [5], mesenchymal stem cells [6,7], amni-
can also be accelerated by environmental factors that otic membrane [8], placental extract [9], cord blood
cause oxidative stress [1,2]. In the male reproductive serum [10], and amniotic fluid [11]. Dried placenta is
system, the aging process can lead to infertility. also rich in protein, fiber, fat, minerals (e.g., sodium,
Semen volume and sperm motility and concentra- potassium, phosphorus, calcium, iron, magne-
tion decline as age increases [3]. A decrease in the sium, zinc, copper, and manganese), and hormones
quality of bull semen due to aging is a problem at (e.g., estradiol, progesterone, testosterone, and growth
artificial insemination centers in Indonesia [4]. Older hormone) [12]. In traditional Chinese medicine, dried
bulls often are retained because the cost to replace human placenta is one ingredient used to treat infer-
them is high. Supplementation to improve bull repro- tility in humans [13]. Compared to human placenta,
ductive performance is an option, but it has not been bovine placenta is easier to obtain in large quantities.
done because limited research exists on regenerative However, dried bovine placenta has never been stud-
agents for male livestock reproductive degeneration ied for its nutritional content or its potential effects as
caused by aging. a regenerative agent in infertility caused by aging.
Copyright: Prihatno, et al. Open Access. This article is distributed Therefore, in this study, we describe the basic
under the terms of the Creative Commons Attribution 4.0
International License (http://creativecommons.org/licenses/
components of dried bovine placenta of beef cat-
by/4.0/), which permits unrestricted use, distribution, and tle from a traditional farm in Yogyakarta, Indonesia.
reproduction in any medium, provided you give appropriate credit
to the original author(s) and the source, provide a link to the
We analyzed the placenta for its potential effects as a
Creative Commons license, and indicate if changes were made. regenerative agent in a rat model of male reproductive
The Creative Commons Public Domain Dedication waiver (http://
creativecommons.org/publicdomain/zero/1.0/) applies to the data
degeneration caused by aging. We used D-galactose
made available in this article, unless otherwise stated. as an inducting agent for male reproductive aging,
Veterinary World, EISSN: 2231-0916 1602Available at www.veterinaryworld.org/Vol.14/June-2021/25.pdf
adapting a method previously described by Salman was separated from the testicular organ. As much as,
et al. [14]. 10 μL of semen from the cauda epididymis was taken
This study aimed to describe the basic compo- using a micropipette. Semen was mixed with 990 μL
nents of dried bovine placenta powder and its poten- of NaCl solution in a 1.5 mL tube. The semen solu-
tial effects as a regenerative agent in a rat model of tion was diluted again using a leukocyte pipette 101.
male reproductive aging with D-galactose induction. Spermatozoa counting was done using a Neubauer
Materials and Methods chamber (Hemocytometer: ASSISTANT®, Germany).
The calculation was carried out 3 times, and the results
Ethical approval
were averaged.
The study was approved by the Research
Ethics Committee, Faculty of Veterinary Medicine, Testes weight and volume
Universitas Gadjah Mada with Ethical Clearance The removed testes were weighed using a dig-
Letter Number: 0023/ EC-FKH/Int./2020. ital scale (model: EHA401, CAMRY®, Zhongshan,
Guangdong, China). After that, the testes were fixed
Study period and location
in Bouin’s solution for 24 h. For preservation, the tes-
This study was conducted from June to November
tes were moved in 70% alcohol solution. The length
2020 at Practical Animal Room and Laboratory of
(l), height (h), and width (w) of the testes were mea-
Reproduction and Obstetrics, Faculty of Veterinary
Medicine, Universitas Gadjah Mada, Yogyakarta sured using a Vernier caliper (Tricle Brand®, Shanghai,
Indonesia. China). Testes volume (V) was measured using the
4
Dried bovine placenta following formula: V = π lhw .
3
Fresh bovine placenta that was expulsedAvailable at www.veterinaryworld.org/Vol.14/June-2021/25.pdf
Table-1: Contents nutrient analysis of dried bovine placenta powder.
Samples Content of dried bovine placenta powder
Water (%) Ash (%) Fat (%) Protein (%) Carbohydrates (by different %)
Bovine placenta powder 1 8.21 (8.25) 5.57 (5.86) 3.32 (2.79) 62.94 (63.84) 19.96 (19.26)
Bovine placenta powder 2 9.05 (8.79) 4.14 (4.16) 3.43 (3.53) 61.49 (59.65) 21.89 (23.87)
Average 8.58±0.41 4.93±0.91 3.27±0.33 61.98±1.83 21.25±2.07
Table-2: The number of spermatozoa from the caudal fetal membrane is normally expelled within 12 h after
epididymis.
delivery of the calf.
Rat number Spermatozoa count (106/mL) Analysis of the dried bovine placenta powder
Group A Group B Group C
showed that it was still rich in nutrients. This find-
ing is in line with a report by Phuapradit et al. [12]
Rat 1 3466 1266 2266
Rat 2 3200 1200 3266
on the nutrients and hormones in a heat dried human
Rat 3 2600 1532 2666 placenta, which was rich in protein and minerals.
Rat 4 2932 2000 - However, the protein hormone concentration was
Rat 5 2932 1466 2732 reduced during the steaming and dehydration pro-
Average 3026±25 *1492.8±315 2732.5±411 cesses [18]. At least 16 hormones can be detected in
*pAvailable at www.veterinaryworld.org/Vol.14/June-2021/25.pdf
4,00,00,00,000 hormone concentrations in dried bovine placenta need
3,50,00,00,000 to be investigated further.
Statistical analysis of testicle weight and vol-
Spermatozoa count
3,00,00,00,000
2,50,00,00,000
ume of rats in Groups A, B, and C showed no sig-
2,00,00,00,000
nificant differences, although the average sperm
*
concentration from the cauda epididymal among the
1,50,00,00,000
groups was significantly different. The average tes-
1,00,00,00,000
ticle weight and volume of the rats in Group B were
50,00,00,000
slightly higher than those (Figures-2 and 3) of the
0
A B C rats in Groups A and C. Similar results have been
Group
previously reported by Amini et al. [40], indicat-
ing that no significant differences were found in the
Figure-1: Concentration of spermatozoa from cauda testicular weight and volume of rats fed a sesame
epididymal semen of rats in Group A, Group B, and Group
C. Statistical analysis showed significant difference (*) with
seed diet although the mean cell number in the left
pAvailable at www.veterinaryworld.org/Vol.14/June-2021/25.pdf
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