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Abstract

The objective of this study was to assess the acute oral toxicity of AV/SGB/27 (M/s Ayurvet Limited) in accordance with OECD-423 guideline. AV/SGB/27 is a herbal formulation for enhancing semen quality of livestock. Nine healthy and adult nulliparous nonpregnant Swiss albino female mice, weighing 24-28 g, were used in this study. After being given test material orally, the mice were examined for toxic effects and mortality. Changes in body weights, symptoms of toxicity, histological appearances of liver, kidney, and lungs, and biochemical parameters were used to assess the toxicity. No toxic effects or mortalities were noticed until the trial was completed, indicating that AV/SGB/27 is safe for oral consumption.
496
The Journal of Phytopharmacology 2021; 10(6):496-499
Online at: www.phytopharmajournal.com
Research Article
ISSN 2320-480X
JPHYTO 2021; 10(6): 496-499
November- December
Received: 30-10-2021
Accepted: 16-12-2021
©2021, All rights reserved
doi: 10.31254/phyto.2021.10611
Sunil Hajare
Post Graduate Institute of Veterinary
and Animal Sciences (PGIVAS), Akola
- 444104, India
Ranjit Suresh Ingole
Post Graduate Institute of Veterinary
and Animal Sciences (PGIVAS), Akola
- 444104, India
Mahesh Kumar Vitthal Ingawale
Post Graduate Institute of Veterinary
and Animal Sciences (PGIVAS), Akola
- 444104, India
Vivek Borekar
Post Graduate Institute of Veterinary
and Animal Sciences (PGIVAS), Akola
- 444104, India
David Kumar
Research and Development Unit,
Ayurvet Limited, Baddi - 173205, India
Ravikanth Kotagiri
Research and Development Unit,
Ayurvet Limited, Baddi - 173205, India
Bhaskar Ganguly
Research and Development Unit,
Ayurvet Limited, Baddi - 173205, India
Correspondence:
Dr. David Kumar
Research and Development Unit,
Ayurvet Limited, Baddi - 173205, India
Email: clinical01@ayurvet.in
Evaluation of acute oral toxicity of a herbal semen quality
enhancer
Sunil Hajare, Ranjit Suresh Ingole, Mahesh Kumar Vitthal Ingawale, Vivek Borekar, David Kumar*,
Ravikanth Kotagiri, Bhaskar Ganguly
ABSTRACT
The objective of this study was to assess the acute oral toxicity of AV/SGB/27 (M/s Ayurvet Limited) in
accordance with OECD-423 guideline. AV/SGB/27 is a herbal formulation for enhancing semen quality
of livestock. Nine healthy and adult nulliparous nonpregnant Swiss albino female mice, weighing 24-28
g, were used in this study. After being given test material orally, the mice were examined for toxic
effects and mortality. Changes in body weights, symptoms of toxicity, histological appearances of liver,
kidney, and lungs, and biochemical parameters were used to assess the toxicity. No toxic effects or
mortalities were noticed until the trial was completed, indicating that AV/SGB/27 is safe for oral
consumption.
Keywords: Acute oral toxicity, AV/SGB/27, OECD 423, Semen quality, Herbal.
INTRODUCTION
Profitability in any commercial livestock-breeding unit is closely related to the reproductive efficiency of
animals. Fertility of the breeding males is important to reduce production losses and to achieve better
economy of the farm. Many herbs have the ability to enhance vigor and fertility in order to improve
reproduction. Low testosterone levels, toxicity causing low sperm count, and poor sperm motility can all
be addressed with herbal therapies. Although the causes of infertility are various and undetermined,
oligozoospermia (low sperm concentration) and asthenozoospermia (low sperm motility) are the most
common causes of male infertility [1, 2]. Hormonal imbalances, mineral deficiencies, rearing stresses, and
radiations are notable factors that lower reproductive performance. So, better semen quality is important
for improving fertility, conception rate and farm economy.
AV/SGB/27 (M/s Ayurvet Limited, Baddi, India) is a scientific blend of effective medicinal herbs and
minerals for improving semen quality, fertility, and vigor of the breeding males. In various studies, its
key ingredients such as Withania somnifera, Mucuna pruriens, Asparagus racemosus, and Tribulus
terrestris, have been used as natural remedies for thousands of years in Indian and Chinese medicine, to
improve sperm quality and erectile functions, promote sexual behaviors and increase hormonal levels [3, 4,
5].
These herbal ingredients promote spermatogenesis by improving the testicular, seminal vesicular and
epididymal functions. They stimulate testicles to produce more and high-quality sperm. They also
improve the sperm count and the quality by boosting the number of LH-FSH-producing basophil cells in
the pituitary. AV/SGB/27 is hormone-free, leaves no residue, and is environment and animal-friendly.
The aim of this study was to determine the acute oral toxicity of AV/SGB/27.
MATERIALS AND METHODS
The present research was performed at the Dept. of Vet. Pharmacology and Toxicology at the Post
Graduate Institute of Veterinary and Animal Sciences (PGIVAS), Akola, Maharashtra. Institutional
Animal Ethics Committee of PGIVAS, Akola, approved the research protocol with approval number
312/4/14/2000/20; dated 06-03-2020.
Nine healthy and adult nulliparous nonpregnant Swiss albino female mice, weighing 24-28 g, were used
in this study. The animals were obtained from the lab animal resource unit, Dept. of Pharmacology,
PGIVAS, Akola, Maharashtra. Animals were cared for in accordance with IAEC SOPs and CPCSEA
guidelines. Picric acid staining was used to identify the animals. For easy monitoring, the number of
animals in single cage was kept at three with good housing conditions. The animals were subjected to
12-hour light and 12-hour dark cycle with maintained 25 ± 20 °C temperature and 70% relative
humidity. All mice were fed a standard pelleted feed and had unlimited access to water [6].
The Journal of Phytopharmacology
497
For acclimatization to laboratory environment, all mice were
maintained in the cages for five days. Thereafter, all mice were fasted
for the night; food but not water was withheld for 3-4 hrs. All mice
were weighed after the fasting period and the test material was given
to them per os. The test sample was given to 3 mice in Group-I with
dose rate of 300 mg/kg of body weight (b.wt.). If signs of toxicity not
appeared in Group-I, then other 6 mice in Group-II were administered
the limit dose of the test materials i.e. at 2000 mg/Kg of b.wt.
Following administration of the test material, no mice were fed for 1-2
hrs in both groups.
The animals were observed for toxic effects and fatalities, as well as
the LD50 value, at least once for the first 30 minutes and frequently
during first 24 hrs, and further for 14 days. Changes in the eyes, skin
and coat as well as changes in the respiratory, circulatory, and central
nervous systems, as well as autonomic and somatic activity were
noted.
Clinical signs like muscular tremors, lethargy, sleep, diarrhea,
salivation, and convulsions, if seen, were noted. Animals were
euthanized after 14 days of examination, and necropsy, along with the
histopathological study of the liver, heart, lungs, and kidneys, were
performed. Biochemical characteristics such as alanine transaminase
(ALT), aspartate aminotransferase (AST), alkaline phosphatase
(ALP), and creatinine were analyzed in the blood samples.
The data of biochemical parameters were analyzed statistically using
one way ANOVA followed by complete randomized design.
RESULTS AND DISCUSSION
The b.wt. of mice were measured individually during 0, 7, and 14
days of the study. The b.wt. of Group-I and Group-II continued to
increase during the study period (Table 1). No mortality and abnormal
sign was seen after oral administration of AV/SGB/27 at 300 mg/Kg
b.wt. and 2000 mg/Kg b.wt. to G-I and G-II mice, respectively. The
LD50 of AV/SGB/27 was more than 2000 mg/Kg as no mortality
occurred at this limit dose i.e. the limit dose, which can be given by
orally.
Necropsy after 14 days did not showed any significant changes in the
gross appearance of liver, heart, lungs, and kidneys. Similarly, no
changes were seen in the histopathological appearances of liver,
kidneys, heart, and lungs in any of the animals (Figure 1). While
blood biochemical parameters differed significantly in the values of
AST, ALT, and creatinine between G-I and G-II (Table 2), the values
were well within their normal ranges in both the groups, indicating no
liver or kidney damage.
AV/SGB/27 is prepared from parts of plants like Withania somnifera,
Mucuna pruriens, Asparagus racemosus, and Tribulus terrestris,
which belong to the Generally Regarded as Safe (GRAS) category.
The extracts of T. terrestris, W. somnifera and M. pruriens regulating
Nrf2/HO1 and NF-κB pathways and enhance sexual activity and
behavior by increasing the androgen levels and simultaneous lower
the ROS levels [5]. W. somnifera, having active component like
sitoindosides and withaferin A [7, 8], reduce lipid peroxidation by
elevation of superoxide dismutase, free radical scavenging enzymes,
catalase, and glutathione peroxidase levels [7]. The active component
in T. terrestris is protodioscin, which improves semen quality by
increasing androgen level and enhances ejaculation [9, 10]. A.
racemosus and M. pruriens contain active ingredients like shatavarin
I-IV and mucunine, which have anti-oxidant and immunomodulatory
activity [ 11, 12]. M. pruriens significantly increases sperm motility and
sperm concentration. Further, it decreases major anomalies of the
sperm [5, 13]. Moreover, M. pruriens increases erectile function, sexual
desire and overall satisfaction and orgasmic function as well as
increases semen volume, total sperm count, and rapid linear
progressive sperm, and decreases non-progressive and immotile sperm
count [14]. Similarly, A. racemosus is useful for infertility and
decreased libido [3]. Thus, AV/SGB/27 can be used to improve semen
quality and infertility problem in male animals without exerting any
toxic effects.
Table 1: Individual weekly body weights and mortality of mice during experimental period
Dose
Animal number
Body weight (g) on day
Mortality
0
7
14
300 mg/Kg b.wt. orally
(G-I)
1
24
26
26
No
2
27
29
29
-
3
28
28
30
-
Mean ± SD
26.33 ± 1.20
27.67 ± 0.88
28.33 ± 1.20
2000 mg/Kg b.wt. orally
(G-II)
1
25
26
27
No
2
26
27
28
-
3
26
27
29
-
4
27
28
28
-
5
27
27
29
-
6
28
29
30
-
Mean ± SD
26.50 ± 0.43
27.33 ± 0.42
28.33 ± 0.43
The Journal of Phytopharmacology
498
Table 2: Mean ± SD ALT, AST, ALP and creatinine values of experimental animals
ALT (U/L)
AST (U/L)
ALP (U/L)
Creatinine (mg/dL)
42.34±0.59b
49.71±0.84b
119.01±2.63
0.45±0.012b
45.72±0.85a
55.18±0.58a
125.59±2.09
0.55±0.012a
a- b Mean ± SD bearing different superscripts differ significantly (p<0.05) within columns
A
B
C
D
Figure 1: Histopathological appearances of A-heart, B-kidneys, C-liver and D-lungs of mice receiving AV/SGB/27 (2000 mg/Kg b.wt.)
CONCLUSION
AV/SGB/27 produced no acute oral toxicity, as indicated by the
absence of mortality, the absence of clinical signs, and the absence of
gross or histopathological changes, even when given upto the
threshold dose in mice (2000 mg/Kg b.wt.). Based on the results, the
herbal medicine is safe to take orally.
Acknowledgments
The authors are grateful to M/s Ayurvet Limited, Baddi, India, for
financial assistance.
Conflict of Interest
None declared.
Financial Support
None declared.
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HOW TO CITE THIS ARTICLE
Hajare S, Ingole RS, Ingawale MKV, Borekar V, Kumar D, Kotagiri R, et
al. Evaluation of acute oral toxicity of a herbal semen quality enhancer. J
Phytopharmacol 2021; 10(6):496-499. doi: 10.31254/phyto.2021.10611
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