EFFECT OF NEEM {Azadirachta inclica A. Juss) EXTRACTS ON GROWTH AND REPRODUCTION OF ALBINO MICE
By
Author
Presented To
Department of Zoology
ABSTRACT
The present study entitled, "Effect of neem (Azadirachta indica A.
Juss) extracts on growth and reproduction of albino mice" was initiated to
have detailed comparative account of the effects of different doses of neem
extracts (leaf, bark and neem oil) on reproduction in male albino mice. Three
different doses of leaf extract (132, 200 and 300mg/kg bw/day), two different
doses of bark extract (200 and 300mg/kg bw/day) and three different doses of
neem oil (2.0, 3.3 and 4.6 ml/kg bw/day) were administered orally for 24 days.
The different doses of neem extracts (leaf, bark and neem oil) when
administered to the mice, affected food consumption, body weight and
weights of testes and accessory reproductive glands such as epididymides
and seminal vesicles. Histomorphological changes in testes and accessory
reproductive glands were also observed after the administration of neem
extracts. Maximum antispermatogenic and antifertility effect was observed
after the administration of leaf extract at the dose level of 300 mg/kg bw/day
for 24 days. Significant decrease was observed in different sperm parameters
such as sperm count, per cent sperm motility, live sperms and normal sperms
after the administration of neem extracts. Fertility rate/index was significantly
decreased in treated mice. It reduced from 100 per cent in control (untreated
mice) to 66, 50 and zero per cent in mice treated with leaf extract at the dose level of 132, 200 and 300 mg/kg body weight. Proteins, deoxyribonucleic acid,
ribonucleic acid, phospholipid levels and acid phosphatase activity decreased
while total lipids, total cholesterol and alkaline phosphatase activity increased
in testes of treated mice. Concentration of testicular trace elements such as
Zn, Cu, Ca, Fe, Na and K was also affected in treated mice. Administration of
leaf extract to immature mice (6 weeks old) at the dose level of 300 mg/kg
bw/day for 28 days did not affect the growth of albino mice however, the body
weight, weights of testes and accessory reproductive organs and various
sperm parameters of cauda epididymis were affected.
Keywords: Neem extracts, growth, reproduction, albino mice.
Abbreviations
AHP
AHS "
CT
DH
EH
ES
HL
EH,
its
Lu
M
MP
MPB
mpg
PS
Py
EHr
RLu
RS
SC
Sp
Spg
ss
ST
Tu
Vc
Abnormal head position
Abnormal head shape
Coiled tail
Double headed spermatozoon
Epithelial height
Elongated spermatid
Headless spermatozoon
Increased epithelial height
Intertubular space
Lumen
Metaphase
Maturation phase
Midpiece bending
microphotograph
Primary spermatocyte
Pycnotic
Reduced epithelial height
Reduced lumen
Round spermatid
Sertoli cell
Spermatozoon
Spermatogonia
Secondary spermatocyte
Seminiferous tubule
Tubule
Vacuole
CONTENT
Chapter Title Page No.
CHAPTER ONE -INTRODUCTION 1-4
CHAPTER TWO - REVIEW OF LITERATURE 5-48
CHAPTER THREE - MATERIALS AND METHODS 49-65
CHAPTER FOUR - RESULTS AND DISCUSSION 66-180
CHAPTER FIVE - SUMMARY 181-185
REFERENCES 186-212
Chapter I
INTRODUCTION
RodcnT S hav e aivVSy'S u66 n with us, mostly as pes lS, SVoT SiHOG Li iS
primitive man became an agriculturist and started having granaries. They
have become a global problem as they damage standing crops, stored food
grains and other commodities. Rodents being the most destructive vertebrate
pests, pose a great threat to our country's economy. Percentage damage by
rodents to various agricultural crops varies under different agroclimatic
conditions (Malhi and Parshad 1990; Islam et al 1993; Jain et al 1993). They
serve as vectors of several diseases (Neelanarayanan and Kanakasabai
2000). They may also gnaw at insulating material causing electrical
installations out of order and creating fire hazards. Rodent problem is very
serious due to their high rate of reproduction, complex behaviour and ability to
adapt under diverse ecological conditions.
Several traditional and modern approaches of rodent
control/management are being used (Parshad 1999). These include non-
lethal approaches like environmental and cultural methods (harbourage
reduction, food source reduction and immigration reduction), cultural practices
(crop rotation, tillage, varietal resistance and crop scheduling) and biological
methods (the use of predators, parasites and pathogens). Lethal or reductional approach includes mechanical methods such as hunting, killing,
trapping of rodents and use of chemicals. However, at present there is no
single effective method that can be applied for all pest species under different
ecological conditions as the response of different species to different
management strategies is very specific. So, there is need to adopt an
integrated approach using all types of control/management methods. One of
the management methods in Integrated Pest Management (iPM) being in
practice is through regulation of their breeding activity (reproduction) by
chemicals (antifertility, antispermatogenic, spermicidal, antiimplantation and
resorptive) (Sinha et al 1984a,b; Upadhyay et al 1993b; Kaur and Parshad
1997). A number of synthetic compounds have been reported to arrest
spermatogenesis or induce 'functional sterility' in rodents (Kaur et al 1997;
Singla and Parshad 2000; Cheng et a/2001; Grima et a/2001). Indiscriminate
use of synthetic chemicals/pesticides have created so many problems like
pesticide resistance, contamination of food, resurgence of pest species and
acute effects on non-target species. The high cost of these chemicals have
generated the need for intensified research on environment friendly chemicals
of plant origin i.e. phytochemicals (Kasturi et al 1995; Sharma and Dixit 1998;
Verma et al 1998; Mazaro et al 2000).
India's rich biodiversity has a vast source of natural products and there
is considerable scope for developing new and improved fertility regulating
agents of plant origin. Extracts of many plants are known to possess antifertility activity (Pakrashi and Pakrasi 1977; Murugavel and A..»jarsha
1991; Raji and Bolarinwa 1997; Aladakatti and Ahamed 1999; Sarkar et al
2000).
Neem (Azadirachta indica) is an outstanding example of tree species,
a versatile gift of nature (Rawat 1995; Kapoor 2000) having antifertility
properties (Tripathi 1998). Tandan et al (1995) and Gupta (1997) reported
that Azadirachta indica possesses various pharmacological activities. Every
part of neem tree namely leaf, flower, fruit, seed kernels, bark, wood, twig and
root, etc. has been used in Ayurvedic and Unani systems of medicine
(Pandey 1993; Rahman and Jairajpuri 1993).
Neem extracts/products have been found to possess antifertility effects
in females (Sharma and Saksena 1959a,b; Sinha et al 1984a; Khare et al
1984; Lai et al 1986; Tewari et al 1986; Riar et al 1988; Upadhyay et al 1990;
Bardham et al 1991; Chattopadhyay 1993; Juneja and William 1993; Dhaliwal
et al 1998; Mukherjee et al 1999).
A few studies were carried out to demonstrate antifertility effects of oral
administration of neem leaf extracts in male rats (Choudhary et al 1990;
Shaikh et al 1993; Kasturi et al 1995; Joshi et al 1996; Parshad et al 1997;
Aladakatti and Ahamed 1999). No detailed study has been made till now to
establish the antifertility effect of oral administration of bark extract and neem
oil in a single rodent species.
Therefore, the present study entitled "Effect of neem (Azadhirachla
indica A. Juss) extracts on growth and reproduction of albino mice" has been
made* on male albino mice Mus musculus to know the effects of oral
administration of neem extracts (leaves, bark and seed oil) on:
i) Histomorphology of the testes
ii) Spermatogenic activity at different cellular levels
iii) Histomorphological changes in accessory reproductive organs
iv) Sperm parameters
v) Fertility index
vi) Testicular biochemistry
vii) Changes in testicular trace elements
viii) Growth
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