08

Review Paper

ANTIFERTILITY ACTIVATES OF TRADITIONAL MEDICINAL PLANTS IN

MALE WITH EMPHASIS ON THEIR MODE ACTION: A REVIEW

Pawan Kumar Soni, Gourav Luhadia, Deepak Kumar Sharma and Pratap Chand Mali

Reproductive Biomedicine and Natural Product Lab.,

Reproductive Physiology Section,

Centre for Advanced Studies, Department of Zoology,

University of Rajasthan, Jaipur -302004.

Abstract

Besides of rapid progress and development of science in medicine field, faith in

and popularity of traditional methods and natural products have not decreased.

There are a huge number of studies which supports the antifertility effects of

plants and their product. The present Study involves various natural plant drugs

and their bioactive component which have antifertility property in male. This

review may help to identify medicinal plants responsible for antifertility activity.

Key words: Antifertility, medicinal plant, male, animal.

INTRODUCTION

In the developing countries like India over population is one of the most serious problems and

world population would be reached about 9.2 billion by the year 2050. The last Indian census

[2011] revealed that the population of India in 2011 was 1,210,193,422. Over population effects

on poverty, environmental degradation, depletion of Natural resources, rise in unemployment.

Modern reproductive biomedicine has provided several preventive and effective methods of

contraceptives for fertility control in male and female but none of which is very safe and without

any serious side effects[1].

Contraception is literally the prevention of conception, but generally is taken to mean the

prevention of pregnancy. Epidemiological studies indicate that combined oral contraceptives

increase risks of cerebral thrombosis, increase serum level of triglyceride, HDL and cholesterol

and increase family mortality due to cardiovascular diseases[2] as well as malignant tumors in

any organs, poor glucose tolerance or diabetes, nausea, abdominal pain, headache, obesity and

menstrual changes [3].Plants were important sources of novel pharmacologically active

compounds, with many blockbuster drugs being derived directly or indirectly, which have been

found in vitro to have antimicrobial [4] antifungal, antiaging and antiimmflamatory properties.

Beside of these, Plants that have antifertility activity [5] and abortifacient properties may act

through rapid expulsion of the fertilized ova from the fallopian tube, inhibition of implantation

due to a disturbance in estrogen - progesterone balance, fetal abortion, perhaps due to lack of

supply of nutrients to the uterus and the embryo, and also on the male side through affecting

sperm count, motility and viability.

Male contraceptive may be work as [6]

1. antispermatogenic agents to suppress sperm production.

2. Prevention of sperm maturation.

3. Prevention of sperm transport through vas deferens.

Journal of Global Biosciences ISSN 2320-1355

Volume 4, Number 1, 2015, pp. 1165-1179

Website: www.mutagens.co.in

E-mail: [email protected]

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Journal of Global Biosciences Vol. 4(1), 2015 pp. 1165-1179 ISSN 2320-1355

http://mutagens.co.in 1166

4. Prevention of sperm deposition.

There are several types of contraceptives which are providing a significant amount of

protection from sexually transmitted diseases [7], and unwanted pregnancies.

To provide modern synthetic medicines for poor and illiterates people is very difficult task. Thus

traditional medicine is often the only affordable and accessible form health care. Therefore, The

use of plants products as anti fertility agent cause minimal side effects as compared to currently

available contraceptive methods. Few herbal contraceptives have been developed but the

potentiality of these contraceptives is very minimal [8]. Several plant bioactive products are

reported to target the testes at the hormonal level or spermatogenesis or both. In this review,

we discuss on some of the commonly used plant products that could be effect on function of the

testis, ultimately cause infertility.

Spermicidal agents

Spermicidal agents are defined as drugs that have the ability to immobilize or kill the sperm

upon contact. An ideal spermicide should immediately and irreversibly produce

immobilization of the sperm, non- irritating to the vaginal and penile mucosa, not have adverse

effects on the developing fetus, free from long- term topical and systemic toxicity and should

not be systemically absorbed. Therefore, one of the oldest approaches for achieving

spermicidal action has been to modify vaginal pH. The acidic pH of lemon juice may

immobilize sperm by denaturing dyein ATPase [9]. Saponins are natural surfactants widely

occurring in many plants (Phytolacca dodecadra, Calendula officinalis and Acacia caesia) are

reported to have spermicidal action. A common lipid bilayer, which contains external, internal

and transmembrane proteins, is fundamental feature of the plasma membrane of the sperm.

Saponin molecules interact with this lipid bilayer, affect the glycoproteins of the cellular

membrane and modify the ionic transport across the membrane, leading to surface changes

[10].

Saponins isolated from Sapindus mukorosii (reetha) are derivatives of hederagenin. At

higher concentrations of saponins (1-50 mg/mL) spermatozoa represented marked

disruption, vacuolation, vesiculation, erosion of the membrane covering the head region and

Coiling of the tail [10].These saponins were formulated into a contraceptive cream named

'CONSAP'.

Some synthetic compounds are also show spermicidal activity as P-nonyl phenoxy polyethoxy

ethanol (N-9) [11], Long Acting, Sustained Release of Spermicide (LASRS) [12],

Benzalkonium chloride [13], Benzalkonium bromide [14]. The active antiliquefying agents as

tannic acid, nitrophenols, and sodium naphthyl phosphate immediately coagulate ejaculated

semen, possibly through a denaturing effect on the glycoproteins present in coagulated

material [15]. 2',4'-dichlorobenzamil hydrochloride [Benzamil] inhibit Na+- Ca2+ exchanger

and Ca2+-ATPase pump present at membrane of the sperm[16].

Spermicidal activity

Botanical name Common name Family Plant part extract Ref.

Acacia

auriculiformis

Ear leaf Acacia Fabaceae Acaciaside A and B 17

Achyranthes Aspera Prickly Chaff

Flower

Amaranthaceae 50% ethanolic extract 18

Aegele marmelos Bael, Golden

Apple,

Rutaceae Leaves extract 19

Albizzia lebbek Benth. Hassk.

siris

Fabaceae Lebbekanin-E. 20

Albizzia procera Fabaceae Oleanolic acid saponin and 21



Seeds. karak,kalsis,safed

siris, karanji

proceric acid saponin

mixture

Allium sativum Lahsun Lilliceae Bulb Crude aqueous

extract

22

Anagallis arvensis red pimpernel,

poor man's

weather-glass

Primulaceae Whole plant 23

Azadirachta indica Neem Meliaceae Leaves Alcoholic extract 24

Cananga odorata Ylang Yalang Annonaceae Root extract 25

Cestrum parqui Chilean Cestrum Solanaceae leaf extract 26

Chenopodium album Lamb's-Quarters Chenopodiaceae seed extract 27

Chrysophyllum

albidum

White Star Apple Sapotaceae Root and bark ethanolic

extract

28

Colebrookia

oppositifolia

Indian Squirrel

Tail

Lamaceae Leaf extract 29

Juniperus phoenicea Cedar Cupressaceae Fruit extract 30

Molluga

pentaphylla

Five Leaved

Carpetweed

Molluginaceae 31

Quassia amara Amargo, Simaraubaceae Stem extract 32

Sapindus mukorosii Reetha Sapindaceae Saponins 33

Sarcostema acidum Sansiveria, Asclepiadaceae Stem extract 34

Terminalia chebula Chebulic Myrobala Combretaceae Fruit extract 35

Tinospora cordifolia Guduchi Menispermaceae Stem extract 36

Trigonella foenum-

graecum

Methi Fabaceae dried seed extract 37

Zizyphus

mauritiana

Ber Rhamnaceae Aq and methanolic bark

extract

38

ANTISPERMATOGENIC ACTIVITY

Spermatogenesis is a complex process which consists [a]Spermatocytogenesis,

[b]Spermatidogenesis [c] Spermiogenesis.

In spermatocytogenesis, a diploid spermatogonium, divides mitotically, producing two diploid

intermediate cells called primary spermatocytes. Each primary spermatocyte undergoes

meiosis I to produce two haploid secondary spermatocytes. Spermatidogenesis is the creation of

spermatids from secondary spermatocytes. Spermiogenesis, includes the differentiation of

spermatids into mature sperm. Spermiogenesis incorporate polarization of the spermatid,

formation of acrosomal cap and flagellum, cytoplasmic remodeling elongation of the nucleus,

develop a thickened midpiece and begin to grow a tail. Antispermatogenic activity as the

number of spermatocytes, spermatids and spermatozoa decreased. Increase in cholesterol

level and Sudanophilic lipid accumulation indicates inhibition in the steroidogenesis.

The extract of Cannabis was reported to induce increased lipid peroxidation in the testis,

along with concomitant decrease in the levels of antioxidant enzymes such as superoxide

dismutase, catalase and glutathione peroxidase. Damage to the basement membrane,

shrinkage of the seminiferous tubules, scanty cytoplasm and shrunken nuclei in the

germinal epithelium, and complete arrest of spermatogenesis were also reported [39]. The

aqueous leaf extract of Azadirachta indica damaged the seminiferous tubules, resulting in the

slackening of germinal epithelium, marginal condensation of chromatin in round

spermatids, degeneration of germ cells and the derangement of germ cell types from their

orderly arrangement in spermatogenesis[40, 41]. Gossypol, a polyphenolic compound

present in the stem, seeds and roots of Gossypium species causes spermatogenesis inhibition

[42].The chloroform extract of Carica papaya inhibit sperm concentration, membrane damage in

the acrosome, bent mid piece, coiled tail, detached head and arrest of spermatogenesis [43].

Piperine, an alkaloid extracted from the fruits and roots of Piper longum, has been shown to



cause damage to the germ cells, increase in lipid peroxidation in testis and epididymis,

inhibition in antioxidant enzymes, activation of caspase 3 and Fas apoptotic protein[44,45].

The extract of Vinca rosea has been Changes in the seminiferous tubule and a decline in the

percentage of primary spermatocytes [46]. Ocimum sanctum has been also reported to cause

to antifertility by targeting spermatogenesis [47]. The rhizome extract of Curcuma longa show

reversible effect and caused reduction in the diameter of seminiferous tubules, loosening of the

germinal epithelium , intraperitoneal vacuolation and mixing of spermatids at different stages of

spermatogenesis in male Wistar rats [48].

Antispermatogenic activity

Botanical name Common name Family Plant extract Ref.

Aegle marmelos Bael Rutaceae Methanolic Leaf-extract 49

Albizzia lebbeck Siris fabaceae Methanolic Pods Bark

extract

50

Allium sativum Lahsun Amaryllidaceae Bulb-Crude Aq extract 51-53

Azadirachta indica Neem Meliaceae Leaves extract 54

Bacopa monnieri Brahmi Scrophulariaceae Leaves- extract 55

Barleria prionitis Vajra-Danti Acanthaceae Methanolic Root- extract 56

Cannabis sativa Ganja Cannabinaceae Alcoholic Root- extract 57

Citrullus colocynthis Tumba Cucurbitaceae Ethanolic Root50%

extract

58

Citrullus colocynthis Bitter Apple Cucurbitaceae Fruit-50% ethanolic

extract

59

Colebrookia

oppositifolia

Bhamber Lamiaceae Leaves-Ethanolic extract 60

Crotalaria juncea Sunn Hemp Fabaceae Seeds-Petroleum

ether,benzene and

ethanol extracts

61

Cuminum cyminum Jeera Apiaceae Seed-Ethanolic extract 62

Curcuma longa Turmeric, Haldi Zingiberaceae 50% ethanolic extract 63

Hibiscus

rosasinensis

Gudhal Malvaceae Bark-Methanolic extract 64

Lepidium meyenii Maca - Brassicaceae Root-Ethanolic Extract 65

Leptadenia hastata Cheila [Konsogna] Asclepiadaceae Leaves and stem-Aqueous

extract

66

Momordica

charantia

Karela Cucurbitaceae Seeds-Petroleum ether,

benzene and alcohol

extracts

67

Mondia whitei White's Ginger Apocynaceae Root bark-Aqueous

extract

68

Morinda lucida Brimstonetree Rubiaceae Leaf-Leaf extract 69

Mucuna Urens Sea Beans Cannabaceae Seed-Ethanol extract 70

Ocimum sanctum Tulsi Lamiaceae Leaf-Aqueous extract 71

Parkinsonia aculata Mexican Palo

Verde

Fabaceae Stem bark-Ethanolic crude

extract

72

Piper nigrum Long Pepper Piperaceae, Fruit-Dry powder-Mice- 73

Rosmarinus

officinalis

Rose-Mary Lamiaceae Fruit-Mehanolic extract 74

Ruta graveolens Rue Rutaceae Leaf-Alcoholic extract-Rat 75

Sarcostemma

acidum

Somlata Asclepiadaceae Stem-70% methanolic

extract

76



Semecarpus

anacardium

Bhilawa Anacardiaceae Fruit-Ethanolic extract 77

Tecoma stans Yellow Bells Bignoniaceae Leaves-Ethanolic extract 78

Terminalia bellirica Baheda Combretaceae Fruit-Alcoholic extract 79

Thevetia peruviana Yellow Oleander- Apocynaceae Stem bark-Methanol

extract

80

Plants that affect Sertoli cells

Sertoli cells are columnar in shape having oval- or pear-shaped nuclei and thin mitochondria,

usually have lipofuscin and lipid droplets at the base of their cytoplasm. Sertoli cell number has

long been thought to be stable in adults with no proliferation of Sertoli cells once adult numbers

have been reached. In the rat, Sertoli cell mitosis ceases after 15 days and FSH then takes control

of differentiation of these cells that are essential to support a qualitatively and quantitatively

normal spermatogenesis. A key feature of Sertoli cell structural support for developing germ

cells is the blood testes barrier that resides in tight junctions located between adjacent Sertoli

cells. This structural arrangement creates an immunologic barrier by isolating the more advanced

germ cell types (spermatocytes and spermatids) from the immune system so that their antigens

do not stimulate autoimmunity.

The Sertoli cells play important role in process of spermatogenesis throughout the adult

life. In addition, the rate and quality of spermatogenesis are monitored by the number of

Sertoli cells present [81].Different studies show that many of plant extracts damage the

viability and function of Sertoli cells and ultimately affects on spermatogenesis in different

manner as reduce nuclear and cytoplasmic volume and vacuolization of the Sertoli cells

(Chloroform extracts of the seeds of Carica papaya), disturb the co-existence of Sertoli–

Sertoli/ Sertoli–germ cell (Azadirachta leaf powder), enlarge of the Sertoli cells (leaf extract of

Thespesia populnea)and reduce the cross-sectional surface area of Sertoli cells(Barleria

prionitis, Dendrophthoe falcate, Albizzia lebbeck).

Plants that affect Sertoli cells


Aegle marmelos Stone Apple, Bael, Bael

Fruit Tree

Rutaceae Methanol leaves 82

Albizzia lebbeck Siris tree, Woman's

Tongue, Saras

Mimosaceae bark methanolic 83

Azadirachta

indica

Neem

Meliaceae leaf powder 84

Barleria prionitis Vjradanti Acanthaceae Root extract 85

Dendrophthoe

falcate

Banda Loranthaceae methanolic extracts 86

Thespesia populnea Tulip Tree, Pacific

Rosewood

Malvaceae leaf extract 87

Tinospora cordifolia Guduchi, Amrita Menispermac

eae

methanolic extracts 88

Tripterygium

wilfordii

Yellow Vine Root Celastraceae 89

Plants that affect Leydig cells

Leydig cells are polyhedral in shape, display a large prominent nucleus,

an eosinophilic cytoplasm and numerous lipid-filled vesicles. The neurons of the

hypothalamus synthesize and secrete gonadotropin-releasing hormone, w h i c h i n d u c e s the



production and release of LH and FSH from the pituitary gland. LH causes the synthesis of

testosterone in the Leydig cells of the testis [90].

Numerous plant products are known to target Leydig cells and hinder their functions

asmethanolic extract of Sarcostemma acidum decrease in the number of mature Leydig cells

and an increase in the degeneration of Leydig cell population [91], 50 percent ethanolic

extracts of the roots of Martynia annua caused Leydig cell atrophy and a significant

reduction in the serum concentration of LH and testosterone [92], methanolic extract of

Tinospora cordifolia reduced stem Leydig cell nuclear area and mature Leydig cell

numbers, [93], leaves extracts of Azadirachta indica exhibited a regresson and decrease in

the number of Leydig cells and their nuclear diameter, indicating androgen deficiency and

Atrophy of the Leydig cells [94],seed extracts o f Carica papaya caused pronounced

hypertrophy of pituitary gonadotrophs and degeneration of Leydig cells [95],Palmitine

hydrochloride(isolated from the roots of Berberis chitria) reduce mature and immature Leydig cells

[96], Ethanolic extracts of Colebrookea oppositifolia decrease in the nuclear and cytoplasmic

surface area of Leydig cells [97] and the flower extract of Malvaviscus conzattii caused[98].

Plants exhibit Effects on Leydig cells


Albizia. lebbeck L Woman's Tongue, Fabaceae methanolic bark extract 99

Azadirachta indica Neem Meliaceae leaf extracts 94

Berberis chitria Indian Barberry, Berberidaceae roots extract 96

Calotropis procera Camelweed Asclepiadaceae root extract 100,

101

Colebrookea

oppositifolia

Indian Squirrel Tail,

Bhamber, Bhirmoli,

Lamiaceae Ethanolic extract 97,

102

Garcinia cambogia Malabar Tamarind,

Brindall Berry

Cluciaceae

Ethanolic Seeds extract 103

Malvaviscus

conzattii

Turk's Cap Mallow Malvaceae flower extract 98

Martynia annua Devil's Claws, Tiger's

Claw

Martyniaceae Ethanolic root extract 104

Ocimum sanctum Tulsi Lamiaceae Leaf powder 105

Affect on motility

The passage of the sperm through three segments of the epididymis-caput, corpus and cauda is

very essential for the final maturation of the sperm [106]. The synthesis and secretion of

various proteins by the epididymis, as well as the attainment of various morphological,

biochemical and motile properties during the passage through the epididymis are

fundamental for the fully fertilizing capabilities of spermatozoa.

Different studies show that many of plant extracts affect motility of the sperm, as the

alcoholic seed extracts of Abrus precatorious significantly decreased cauda epididymal sperm

motility and brought about a decrease in the levels of succinate dehydrogenase and ATPase in

the sperm of male albino rats. The SEM studies on sperm morphology revealed decapitation,

acrosomal damage and formation of bulges on the midpiece region of sperms due to administered

of Abrus precatorious seed extracts [107]. The methanol seed extract of Abrus precatorious was

reduce motility due to the decline in cAMP and enhanced production of reactive oxygen

species [108].

The ethanolic extracts of Azadirachta indica to adult male mice at 0.5 mg, 1 mg or 2 mg per kg

b.w. for 6 weeks interfered with sperm DNA and caused chromosome strand breakage,

spindle disturbances and deregulation of genes responsible for sperm morphology and sperm

motility was decrease linearly with various concentrations (1–50 mg per 1 million sperm) of



neem leaf extract, with motility falling to absolute zero within 20 s of exposure to 3 mg dose

[109]. Piperine at 10 mg and 100 mg per kg b.w. Show a reduction in rat sperm motility,

viability and count [110].An in vitro study on the effects of allitridum, an active principle

from garlic, has been reported to inhibit sperm motility and complete immobilization of rat,

hamster and human spermatozoa at a dose of 7.5 mg mL-1 of allitridum treatment [111]. The

benzene leaves extract of Ocimum sanctum at a dose of 250 mg per kg b.w. for 48 days was

demonstrated to decrease sperm count, motility and the forward velocity of the sperm. The effects

were found to be reversible upon withdrawal of treatment for 2 weeks [112].

Plants affect on sperm motility

Botanical name Common name Family Plant extract Ref.

Abrus precatorius Rosary Pea Fabaceae Seed Dry extract 113

Aegle marmelos Bael Rutaceae Leaf-Methanolic extract 114

Aegle marmelos Shreephal Rutaceae Leaf and ethanol extract 115

Bursera fagaroides Elephant Tree Burseraceae Cortex/ethanoic extract. 116

Carica papaya Papita Caricaceae Seeds Chloroform extract 117

Carica papaya Papita Caricaceae Seeds Chloroform extract 118

Cyclamen persicum Cyclamen Primulaceae Saponins compounds 119

Echinops echinatus Utakatira,

Oontkateli

Asteraceae Root/50% alcoholic extract 120

Gossypum

herbaceum

Cotton Malvaceae

Gossypol compound 121,

122

Hedera nepalensis Himalayan Ivy Araliaceae Nepalin compound 123

Lagenaria

breviflora Robert

Molina Cucurbitaceae Whole fruit and ethanol extract 124

Pentapanax

leschenaultii

Araliaceae Fruit/ethanolic extract 125

Phytolacca

dodecandra

Ghana Akan,

Guinea Kissi

Phytolaccaceae BuOH extract of berries 126

Primula vulgaris Primrose Primulaceae Saponins compounds 127

Effect on Testosterone

The spermatogenesis is regulate by different hormones which are secreate by the

hypothalamus, pituitary gland and testis [128].The major male sexual hormone Testosterone

is synthesized and secreted by Leydig cells under the control of the pituitary gonadotrophin

luteinizinghormone [LH].

Journal of Global Biosciences ISSN 2320-1355

http://mutagens.co.in

Numerous plant products ar

androgens. Gossypol acetic acid

plant when incubated with isolate

dramatic decrease in histochemica

the compound [129]. The crud

in the levels of testosterone, LH

[20 g L−1] and Mentha spicata

LH levels and decreased tota

steroidogenic enzymes including the

dehydrogenase,17α-hydroxylase

was observed when primary mouse Leydig cells were incubated with

crude Toona sinensis [132]. Dose

hydroxysteroid dehydrogenase

Abrus pecatorius was administered

Botanical name Common name

Abelmoschus

esculentus

Okra, Lady's Fingers

Abrus precatorius Coral Bead Vine,

Rosary Pea, Ratti,

Aegle marmelos Bel, Beli Fruit, Bengal

Quince, Stone Apple,

Albizia. lebbeck L Siris

Allium sativum Clove Garlic, Poor

Man's Treacle

Journal of Global Biosciences Vol. 4(1), 2015 pp. 11

are known to target enzymes which are utilize in synthesis of

acid, a polyphenolic compound isolated from

isolated rat interstitial cells at a dose of 50

histochemical stain for 3-β-HSD, proving the direct

crude methanol extract of Quassia amara was reported

H and follicle-stimulating hormone [130]. Mentha piperita labiatae

a labiatae [20 g L−1] was identified to increase

total testosterone levels [131]. Suppression of the activities of

steroidogenic enzymes including the P450 side-chain cleavage enzyme

hydroxylase,20α-hydroxylase and 17 β-hydroxysteroid

when primary mouse Leydig cells were incubated with varyin

. Dose-dependent decrease in the enzyme activity

dehydrogenases and degeneration of Leydig cells wer

administered to male rats [133].

Effect on Testosterone

Common name Family Plant extract

, Lady's Fingers Malvaceae Fruit and

extract

Coral Bead Vine,

Rosary Pea, Ratti,

Fabaceae Dry extract from

Bel, Beli Fruit, Bengal

Quince, Stone Apple,

Rutaceae Leaf and ethanol extract

Fabaceae . Bark and

extract

Clove Garlic, Poor

Man's Treacle

Lilliceae Crude extract

Vol. 4(1), 2015 pp. 1165-1179

1172

enzymes which are utilize in synthesis of

m the seeds of cotton

0 µg mL−1 caused a

t inhibitory effect of

s reported to reduce

Mentha piperita labiatae

increase the FSH and

n of the activities of

enzyme,3β-hydroxysteroid

hydroxysteroid dehydrogenase,

varying concentrations of

activity of 3α, 3β, 17β-

were reported when

Ref.

methanolic 134

from seed 133

Leaf and ethanol extract 135

methanolic 136

137



Bulbine natalensis Bulbine Asphodelaceae Stem and aqueous extract 138

Chromolaena

odoratum L.

Siam Weed,

Christmas Bush, Devil

Weed,

Asteraceae Leaves and aqueous

extract

139

Curcuma longa L Turmeric Zingiberaceae Crude alcoholic extract 140

Dendrophthoe

falcate

Honey Suckle

Mistletoe, Banda

Loranthaceae Stem methanol extract 141

Juniperus

phoenicea L.

Cedar Cupressaceae Cones and ethanol extract 142

Psoralea

corylifolia

Babchi Leguminosae Crude extract 143

Stevia rebaudiana Sweet Leaf, Sugar

Leaf

Astraceae Whole plant, aqueous

extract

144

Syzygium

aromaticum L

Clove Myrtaceae Flower buds and hexane 145

CONCLUSION

Ultimately, it was clear that the herbal medicinal plants and their extracts have significant

antifertility activity in animal models. This review showed that above mentioned medicinal

plants possess antifertility activity on dose dependent manner and can be used as an alternative

for oral contraceptives which are currently in use for birth control.

ACKNOWLEDGEMENT

Authors are thankful to the Head and Coordinator CAS, Department of Zoology, University of

Rajasthan, Jaipur for providing laboratory facilities and the UGC for financial support.

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