Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology

ISSN No:-2456-2165

Antifertility Effect of Aqueous and Alcoholic

Extract of Momordica Dioica and
Melia Azedarach on Male Albino Rats
Richa Sharma1 and D.N.Choudhary2*
1
Research Scholar, 2Associate Professor
University Department of Zoology, TMBU, Bhagalpur, Bihar, India

Corresponding Author: D. N. Choudhary*

University Department of Zoology, TMBU, Bhagalpur, Bihar, India

Abstract:- Plants have long been utilized to treat a from ancient times[7]. Numerous expert have looked at the
variety of illnesses. Search for male antifertility factor of contraceptive qualities of hers and plant components in both
medicinal plants remained a potential area of men and women[8]. It has been noted that many
investigation. The present investigation was aimed to phytochemicals are becoming more significant in man [5].
study the effect of oral administration of two medicinal
plants , Melia azedarach (leaf) and Momordica In emerging nation, the population growth is one of the
dioica(fruit) at a dose of 100mg/kg body weight for 21 main causes of poverty and population. It is becoming a
days on reproductive organs of male albino rats. Body major problem i.e., facing a significant strain on economic,
weight, Reproductive Organ weight , Fructose content social and environmental resources. It is creating so many
in Coagulating gland(CG), Acid phosphatase activity in obstructions worldwide day by day[2][9]. With reference to
Ventral prostate(VP), Total protein in Seminal adjusting the human reproductive rate this overpopulation
vesicle(SV) were investigated in both control and treated can be controlled biologically[11]. Though different
groups. Histology of testis in treated rats showed hormonal contraceptive tablets are being developed
degenerative changes in the seminiferous tubules. The alongside, however, they all have negative effects, hence
decrease in sperm count with wider lumen, organ weight, there is an urgent need for an efficient plant based drugs to
fructose content in CG, protein content in SV, suggesting oppose these problems. In this context use of common
the antifertility activity in both tested plants. Corpus medicinal plants in reproduction meaning may be an
luteal sites, Implantation sites, and Litter size were also important tool to prevent population explosion[2][5].
found significantly reduced. Though anti spermatogenic activity of some medicinal
plants have been reported by many workers[3][7]. Few
Keywords:- Antifertility, Biochemical Parameters, Melia plants also possess male anti fertility activity[12][13].The
Azedarach, Momordicadioica, Sperm Count, Testis. present work has been carried out to study the antifertility
activity of two common medicinal plants i.e., Momordica
I. INTRODUCTION dioica(fruit) and Melia azedarach(leaf) on reproductive
organs of male albino rats. To know the contraceptive
Plants have historically been utilised to treat variety of strength of these two considered or selected medicinal plants
diseases. From time immemorial, humans have relied on rats were treated with both aqueous as well as alcholic
plants that could meet their basic necessities such as food, extract.
shelter, fuel and health[1][2].Plants have been used globally
across varied cultures as a safe natural source of medicines. II. MATERIAL AND METHOD

Search for male antifertility factor of medicinal plants A. Momordica dioica

remained a potential area of investigation[1][3][4]. For Momordica dioica, or spiny gourd, is a perennial
thousands of years Indian traditional medicine has been climber belongs to the Cucurbitaceae family. Traditional
performed through the use of Ayurveda. Many studies on the medical practitioners employ the fruit, leaves, and roots of
pharmacology and clinical therapies of Ayurvedic medicinal Momordica dioicaas medication. Plant extracts are thought
herbs have been conducted. It has been noted that many to provide antidiabetic[14], hepatoprotective[15],
phyto chemicals are becoming more significant in man[5]. renoprotective[16] and anti-hyperglycemic[17]properties.
In numerous animal model plants have been found to have Strong antioxidant properties have been observed for the
therapeutic and contraceptive properties[6].The practice of alcoholic extract of Momordica dioica. According to a
using contraceptive is not new. Contraceptive use has been study, the root of M. dioica extracted in ethanol has potent
expanded dramatically over last 40-50 years among women abortifacient properties[18]. The present study was
of all reproductive ages. Numerous plant and plant derived conducted to assess the antifertility activity of alcoholic and
items have been used as contraceptives and abortifacient aqueous extract of fruit of Momordica dioica.

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
B. Melia Azedarach Male rats was sacrificed on 22nd day and different
The plant Melia azedarach Linn, or mahanimba, is a tissues were collected and weighed on Torsion balance.
member of the Meliaceae family[19]. It is a big, neem-like Serial sections of testis were prepared for microscopic
evergreen tree that grows all over India. It has historically observations [23].
been used as an astringent, stomachic, emmenagouge,
anthelmintic, and antilithic diuretic[20]. The study was Fructose estimation in Coagulating gland (CG) was
conducted to assess the antifertility activity of ethanolic and evaluated by colometry[24]. Acid phosphatase activity in
aqueous extract of leaf of Melia azedarach. Ventral prostate (VP) was evaluated by method adopted
Sigma Technical Bulletin no. 104[25]. Protein estimation in
C. Experimental Animal seminal fluid was estimated by colorimetry[26].
Male Sprague dawleyrats (170-200gm) body weight of
proven fertility were selected for the experiment. Three Spermatozoa collected from Caput, Corpus and Cauda
separate groups ( One for control and two for experimental) epididymis and vas were examined under compound
of male rats were selected . Each group containing 7 microscope and their number and morphology were
animals. The experimental group of rats were administered recorded [2][22].
orally with suspension of test plant extracts at a dose of
100mg /kg Body weight for 21 days .The control group was The data were analysed statistically using student t-
fed with distilled water for the same period of Test.
treatment[2][4].
III. RESULTS
D. Plant Extract Preparation
A. Body weight
 Alcoholic Extract When Momordica dioica fruit extract (aqueous and
The plant extract was prepared by the method alcoholic) was administered orally, there was a significant
adopted[21]. Whole fruit ofMomordica dioicaand leaves of drop (P<0.01) in body weight when compared to the control
Melia azedarachwere taken for investigation. The fruit and group. Similarly, rats showed significant reduction (P< 0.01)
leafof the test plant were washed and dried at room in the body weight after oral administration of Melia
temperature . The dried plant parts were powdered by using azedarach( both aqueous and alcoholic) compared to
mixer grinder. 20gm of plant powder of each test plant was control( Table 1 and graph 1).The pituitary gland may be
poured into a conical flask containing 150ml of 50% of suppressed or has secreted less growth hormone (GH),
ethanol. The mixture was stirred, allowed to settled, and which would explain the cause of lower body
kept covered. At the end of second day the extract was weight[2][27][28].
filtered with no.1 what man filter paper. The filterate was
taken on a petri dish and evaporated at room temperature. B. Reproductive Organ Weight:
The residue remained in the petri dish was ready for There was a significant reduction in weight (P <0.01)
experiment. detected in the reproductive organs like Seminal vesicle
(SV), Coagulating gland (CG),Ventral prostate ( VP) Testis
 Aqueous Extract and Epididymis(Table 1 and graph 2). Significant weight
Whole fruit of Momordica dioicaand leaf of loss of multiple reproductive organs in treated male rats may
Meliaazedarchwas taken. These parts was washed and air be due to decreased gonadotrophic activity or low androgen
dried. For extract preparation, 10g of dried plant parts will levels that interfere with spermatozoa formation and
be dissolved in100ml of distilled water and left for 12hrs maturation[29][30].
,filtered and diluted with distilled water for required
dose(0.1ml/rat/day). After 12 hrs the solution will be C. Biochemical Parameters
filtered. The filterate is now ready for dilution and required When compared to the control group, there was a
dose preparation[2][22]. significant decrease (P<0.01) in the biochemical parameters
such as Protein, Fructose, Acid Phosphatase activity
Fertility performance of individual rat was done from following the oral administration of both the test plants i.e.,
day 16th to 21st of treatment . Each male rat was caged Momordica dioica(fruit) and Melia azedarach(leaf)
separately with 2 coveal female of proven fertility. Presence (aqueous and alcoholic (Table 2 and Graph3). Decline in
of vaginal smear indicated that the females had mated to the fructose content in Coagulating gland (CG) and acid
particular male and the day of mating was considered to be phosphatase activity in Ventral prostate (VP) were possibly
the day 1st of pregnancy. due to decrease secretion of endogenous androgen [2][24].
Reduced protein content in SV of treated rats compared with
Laprotomy was done on 8th day of pregnancy to control group was due to toxic manifestation which lead to
examine the record of Corpora lutea and Implantayion sites. the breakdown of protein and impaired source of ATP
Litters was examined and litters size was recorded at production to meet the energy requirement [31].
term[3][23].

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
D. Haematological Parameters IV. HISTOLOGY OF TESTIS
After the oral administration of Momordica
dioica(fruit) and Melia azedarach(leaf) (aqueous and A microscopic histological analysis of control group
alcoholic) though, there was reduction found in blood testes revealed normal germinal epithelium maturation and
glucose and haemoglobin level as compared to the control cytoarchitecture. Every stage of spermatogenesis ,including
group but it was not significant (Table 3 , Graph 5&6). primary and secondary spermatocytes and germinal
epithelium producing spermatogonia divided by sertoli cells,
E. Sperm Count is indicative of a healthy testis. Mature sperm ready for
Sperm count was also found significantly decreased ( release were visible in the lumen.
P< 0.001 and P< 0.05 ) in rats treated with both the plants
Momordica dioica(fruit) and Melia azedarach(leaf) Testes of rats treated with an aqueous and alcoholic
respectively compared to control group (Table 2 and Graph extract of Momordica dioica(fruit) showed sign of
4).The lowered sperm count in the treated male rats could be deteoriation of spermatogonial layer in the seminiferous
the consequence of disintegration of seminiferous tubule tubules and a decrease in the number of sperm in the lumen.
and leydig cell as well as decreased testosterone Though the tubules showing the trend of degeneration but it
synthesis[30][32]. It is commonly known that testosterone was not significant. Only one or two tubules were found
plays a critical role in spermatogenesis and affects the ruptured in the focus.Wider lumens were present with less
epididymal environment. It must have therefore affected spermatozoa and degeneration in spermatogonial layer were
spermatogenesis. Therefore, it was clear that alcoholic and observed(fig. 4&5).
aqueous extracts of both the test plants have antiandrogenic
property that may lead to antifertility in male The testes of rat treated with an alcoholic extract of
rats[27][30][33]. Melia azedarach(leaf) showed comparatively higher
degeneration in histology as well as number of sperms. The
F. Fertility performance test seminiferous tubules showed wider lumen and less
After oral administration of Momordica dioica(fruit) spermatozoa. Rupture in spermatogonial lining were also
extractMelia azedarach(leaf) extract(both alcoholic and observed under the microscope.The testis or rat treated with
aqueous ) a significant decrease (P<0.01) incorpus luteal aqueous extract of Melia azedarach though showed
sites, implantation sites and litter size, was observed ( Table spermatozoa in the lumen but there are certain
4and Graph 7 ). abnormalities(breakage) found in the spermatogonial lining.
 Momordica dioica : Out of seven, Only five male rats Spermatozoa in the lumen were also found clumped in the
treated with an aqueous fruit extract of Momordica lumen of the tubule in comparison to testis of rats treated
dioica could be able to mate with only five normal with an alcoholic extract of Melia azedarach(fig.1&2).
female rats. In contrast, only three male rats out of seven
treated with alcohololic fruit extract of M. dioica were The reduced testicular weights and ruptured
able to mate with four normal female rats. seminiferous tubule with decreased tubular diameter
 Meliaazedarach : Out of seven, only three male rats indicated wide spread damage[38].A reduction in the tubular
treated with an aqueous leaf extract of Melia azedarach diameter with less number of spermatozoa could be due to
could be able to mate with only three normal female rats a destructive effect of ethanolic extract of Momordica
whereas only four male rats out of the seven rats treated dioica(fruit)and Melia azedarach (leaf).Degeneration of
with alcoholic leaf extract of Meliaazedarcah were able seminiferous tubules and Leydig cells, less diameter of
to mate with only four normal female rats. tubular lumen with insufficient number of spermatozoa
could be due to insufficient production of androgen or
The decrease in implantation sites were obviously due antiandrogenic property of both the test
to changes in endocrine activity of luteal structures. Many plants[32][39].Seminiferous tubules with reduced or
plants have been reported to be anti-ovulatory and anti- ruptured spermatogenic components in both experimental
implantational[34]. However, the extracts showed groups suggested widespread injury to the testicular
abortifacient effects as there was reduction in viable litter structure. There were extremely few or no spermatozoa in
size. These abortifacient effects were indicative of either the lumen of some of the tubules. The testis's seminiferous
changes in maternal estrogen/progesterone ratio or may be tubular diameter (STD) shrank. In the testis of treated rats,
due to either inhibition of implantation or increased loosening and sloughing of the germinal epithelium were
resorption of fetuses[35]or due to some toxic components also noted[40].
reaching the female genital tract with semen[2][3]. In
addition , the number of decreased implantations may also
be due to the decreased sperm count and motility[37]may
be another important reason.

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
ISSN No:-2456-2165
Table 1: Showing the effect of aqueous and Alcoholic Extract of Momordicadioic( fruit)and Melia Azedarach (leaf ) on Body
Weight and Reproductive Organ Weight in Male Albino Rats
Male rats Initial body Final body Wt. of reproductive organs
(Control/traeated) weight weight Testis S.V V.P C.G E.P
Control 190.6±3.5 194.4±4.09 0.18±0.12 0.50±0.02 0.53±0.01 0.17±0.007 0.06±0.014
Momordica dioica 179.4±1.2 180.6±2.83 0.03±0.01 0.30±0.01 0.11±0.01 0.1±0.01 0.01±0.028
(aqueous.)
Momordica dioica 184.4±3.48 188±4.32 0.04±0.008 0.28±0.018 0.11±0.010 0.09±0.005 0.08±0.01
(alcoholic)
Melia azedarach 187.17±4.13 193.63±3.23 0.04±0.01 0.48±0.11 0.08±0.01 0.20±0.01 0.09±0.02
(aqueous)
Melia azedarach 175±1.37 176.8±1.39 0.02±0.008 0.38±0.03 0.08±0.008 0.18±0.02 0.09±0.21
(alcoholic)
Abr. : SV- Seminal vesicle VP- Ventral prostate CG- Coagulating Gland EP- Epididymis.

Table 2: Showing the Effect of Aqueous and Alcoholic Extract of Momordica Dioica( fruit)and Meliaazedarch ( leaf) on Sperm
Count and Biochemical Parameters
Male rat Sperm count Fructose(CG) Protein(SV) Acid phosphatase(VP)
(Control/Treated) mg/100mg of tissue mg/100mg of tissue mg/hr/100mg of tissue
Control 196.9±3.001 0.53±0.01 32.8±0.86 42.26±0.56
Momordica dioica 90.8±1.42 0.16±0.008 13±0.94 15.44±0.66
(aqueous)
Momordica dioica 79±1.41 0.14±0.015 10.2±0.91 10.53±0.967
(alcoholic)
Melia azedarach 89.92±1.49 0.2±0.02 8.67±1.76 17.66±2.52
(aqueous)
Melia azedarach 94.6±1.80 0.02±0.0008 8.6±1.20 12.49±2.12
(alcoholic)

Table 3: Showing the Effects of Aqueous and Alcoholic Fruit and Leaf Extract of Momordica Dioica and Meliaazedarch on
Haematological Parameters
Male rats (control/Treated) Blood glucose(mg/dl) Haemoglobin(gm%)
Control 109±1.18 10.3±0.3
Momordica dioica 104.4±1.96 10.64±0.89
(aqueous)
Momordica dioica 104.4±1.93 8.8±0.33
(alcoholic)
Melia azedarach 109±2.32 6.07±0.77
(aqueous)
Melia azedarach 103.6±1.69 8.0±1.02
(alcoholic)

Table 4: Showing the Effects of Aqueous and Alcoholic Extract of Momordica Dioica(Fruit)and Melia Azedarach(Leaf) on
Fertility Performance Test
Effects of plant extract on fertility of male rats
No. of breeder No. of successful No. of Mated Corpora Implantation Litter
males males females lutea site size
Control 7 5 7 3.9±0.27 2.8±0.29 5±0.31

Momordica dioica 7 5 5 2±0.42 1±0.25 2.5±0.28

(aqueous)
Momordica dioica 7 3 4 2.88±0.39 1±0.26 2±0.40
(alcoholic)
Melia azedarach 7 3 3 4±0.68 2.33±0.67 3.25±1.25
(aqueous)
Melia azedarach 7 4 4 1.67±0.42 1±0.25 2±0.57
(alcoholic)

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
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200

195

190
Body weigh (gm)

185

180

Initial body weight

175
Final body weight
170

165

Groups

Graph 1: Showing the Effect of Both Plant Extracts On Body Weight In Male Albino Rats

Reproductive organ weight

0.6
Reproductive Organ weight (gm)

0.5

0.4
Testis
0.3
S.V
V.P
0.2
C.G
0.1 E.P

0
Control Momordica Momordica Melia Melia
dioica (aq.) dioica (alc.) azedarach (aq.) azedarach (alc.)
Groups

Graph 2: Showing the Effect of Both Plant Extracts On The Reproductive Organ Weight.

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
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Biochemical parameters
60

50

40
Amount

30

20

10

0
Control Momordica Momordica Melia Melia
dioica (aq.) dioica (alc.) azedarach (aq.) azedarach (alc.)
Groups

Graph 3: Showing the Effect of Both Plant Extracts on the Biochemical Parameters (Protein, Fructose, Acid Phosphatase)

Sperm count
250

200
Number in million

150

100
Sperm count

50

0
(no. of Control(7) Momordica Momordica Melia Melia
animals) dioica (aq.) dioica (alc.) azedarach azedarach
(aq.) (alc.)
Groups

Graph 4: Showing the Effect of Both Plant Extracts On The Sperm Count

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Haemoglobin
12

10

8
gm%

4 Haemoglobin(gm%)

0
Control (7) Momordica Momordica Melia Melia
dioica (aq.) dioica (alc.) azedarach (aq.) azedarach
(alc.)
Groups

Graph 5: Showing the Effect of Both Plant Extractson Haemoglobin Test

Blood glucose(mg/dl)
110
109
108
107
106
mg/dl

105
104 Blood glucose(mg/dl)
103
102
101
100
Control (7) M. d (aq.) M. d (alc.) M. a (aq.) M. a (alc.)
Groups

Graph 6: Showing the Effect of Both Plant Extracts on the Blood Glucose

Fertility performance test

8
7
6
5 No. of breeder males
Number

4 No. of successful males

3 No. of mated females
2 Corpora lutea
1 Implantation site
0 Litter size
Control M.d (aq.) M. d M. a M. a
(alc.) (aq.) (alc.)
Groups

Graph 7: Showing the Effect of Both Plant Extracts on the Fertility Performance Test of Male Rats

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Volume 9, Issue 1, January 2024 International Journal of Innovative Science and Research Technology
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V. DESCRIPTION OF HISTOLOGICAL
PHOTOGRAPH

Fig 5: Histology of Testis

Fig. 1: Histology of Testis treated with Aqueousextract of treatedwithAlcoholicextractofMomordica dioica(10X)
Melia azedarach(10X)
VI. CONCLUSION

The present study indicated that oral administration of

both alcoholic and aqueous extract ofMomordica
dioica(fruit) and Melia azedarach(leaf) extract significantly
affectsreproductive functionof experimental rats because it
altered androgen levels andcauses less sperm count and
testes cytoarchitecture.Thus,it may be concluded that both
the tested plants were found to have potential antifertility
agent for decreasing spermatogenic activity.

ACKNOWLEDGEMENT

Fig. 2: Histology of Testistreated with Alcoholic extract of The authors greatefully acknowledge the laboratory
Melia azedarach(10X) facilities provided by the University department of Zoology,
TMBU, Bhagalpur, Bihar. We are also thankful to some
research fellows of the department for their continuous
support. We onvey our gratitude to the teachers of the
department for their valuable suggestions incorporated in
this manuscript.

CONFLICT OF INTEREST

The authors declare that there is no conflict of interest

regarding the publication of this paper.

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Fig. 3: Histology of Testis of Control Rat
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