WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Narayanaswamy et al.
World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 6.041
Volume 5, Issue 7, 1503-1510.
Research Article
ISSN 2278 – 4357
STUDIES ON OVARY OF FRESH WATER FISH GLOSSOGOBIUS
GIURIS (HAM)
Dr. Narayanaswamy S. Y.* and M. Ramachandra Mohan
Department of Zoology, Rural Degree College, Kanakapura.
Article Received on
21 May 2016,
ABSTRACT
The neem oil is a useful local remedy in some chronic forms of skin
Revised on 10 June 2016,
Accepted on 30 June 2016
diseases and ulcers. It is also antiseptic and commonly used for
DOI: 10.20959/wjpps20167-6747
animals both internally and externally. The oil is also extremely useful
as a parasiticide. Fish is a nutritious food for peoples, hence a gonad of
*Corresponding Author
Glossogobius giuris observed during spawning phase on treatment
Dr. Narayanaswamy S.Y.
with sub lethal concentrations of Neem oil (0.05 ppm, 0.25 ppm and
Department of Zoology,
Rural Degree College,
Kanakapura.
0.5 ppm) for 24, 48, 72 and 96 hrs intervals. The large and sexually
mature female fishes were used in this study. The body length and
weight of fishes were recorded. The female fish weighing about 20-50
g and length 110 to 220 mm were selected for ovaries, the ovaries contain ripen eggs
with large oocytes, well defined nuclei, distinct nucleoli and a few immature oocytes. The
results of the present study showed morphological changes in the cell structure, the ovarian
growth was inhibited, the oocytes and interstitial tissues were encapsulated by a connective
tissue, capsule consisting of germinal epithelium and tunica albuginea with blood vessels. As
the concentration increases, the amount of atretic follicles increased and the ovarian wall
became more thickened and lifted. The ovaries of fish exposed to higher concentration an
immense amount of empty cells along with necrosis of nuclei have seen.
KEYWORDS: Neem oil, G. giuris, oocytes, nucleoli and tunica albuginea.
INTRODUCTION
The neem oil is a useful local remedy in some chronic forms of skin diseases and ulcers, by
stimulating and exciting a healthy action. Applied to foul and sloughing ulcers, it retards the
sloughing process to some extent, prevents the production of maggots and dislodges them if
already produced. It is also antiseptic and commonly used for animals both internally and
externally. The oil is also extremely useful as a parasiticide in varius cutaneous affections
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such as ring worm, scabies and others. When the parasite is in the deeper layer skin, it will
be necessary to rub the oil will in for perhaps 10 minutes or more at a time. Internally, the oil
in 5-10 minimum doses, once or twice a day is useful in chronic malarial fevers. Neem oil
has good larvicidal properties for mosquitoes
[1]
. Application of neem to rice fields is
positively evaluated with the dual objective of controlling the culicinemosquite vectors of
Japanese encephalitis virus and enhancing the grain yield
[2]
. Effect of aqueous extrat from
neem on hydrophobicity, biofilm formation and adhesion in composite resin by candida
albicansis proposed [3]. The hexane and alcoholic extracts of neem seed seem to be promising
anticandial agents [4]. A novel use of neem oil, a traditional plant product, for long term and
reversible blocking of fertility after a single intrauterine application is described
[5]
. There is
provided a means for reversible fertility control for prevention of pregnancy in female
mammals for several months through the use of neem oil or its components, applied locally to
the uterus. There is also provided a spermicidal composition containing neem oil or its
components in combination with reetha extracts as a vaginal cream
[6]
. Among the herbal
pesticide, neem oil considerd to be an important pesticide in controlling pests and insects [7,8].
The effect of neem oil its products on fishes have been studied by[9], on
mosquitoes[1,2,10,11,12,13,14], on rats. Arrest of follicle development recrundscences, atresia
ruptured follicles and necrosis
[9]
were noted treatment with neem leaf meal in Tilapia Zilli
(Gervais) fishes.Decreases the number of ovarian follicle, significant changes in the levels of
associated reproductive hormones and presence of higher concentrations of active neem
components in the gonads of female rats. Neem oil shows toxicity of fish like tilapia and
carp[15], in rats and rabbits
[16]
, in humans
[15,17]
. Neem leaf extract when administered for 48
days in albinorats causes decrease in sperm count, sperm molity, probably due to androgen
deficiency
[18]
. Hence, the present investigation, histopathological studies in the ovary of
Glossogobius giuris has been made during spawning phase.
MATERIAL AND METHODS
The fresh water gobiid fish Glossogobius giuris (HAM) were randomly collected in and
around Bangalore using cast and gillnets (10 mm). The fishes were brought alive to the
laboratory and were kept in 50 L aquaria containing aerated tap water and acclimitated in
laboratory conditions for 15 days prior to using them in experiments. They were keep under
natural photoperiod and room temperature of 26 ± 4 ◦C and were fed daily with earth worms.
These fishes were treated with 0.1% potassium permanganate solution for 15 min to get rid of
dermal infections. The large and sexually mature female fishes were used in this study. The
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body length and weight of each fishes were recorded. The female fish weighing about 20-50
g and length 110 to 220 mm were selected for ovaries. The stock solution of 1 mg/L is
prepared and the desired concentration is obtained by adopting the dilution technique outlined
by [19]. The acclimitated fishes were divided into three experimental groups of six fishes each.
The first three groups of fishes are placed in 0.05, 0.25 and 0.5 ppm of neem oil, while fourth
group kept in fresh water (control). Six fishes in each concentration in 10 L capacity glass
trough. For all experiments the acclimitated fishes were starved for 24 hr prior to their
exposure to neem oil use in experiment and were not fed during the course of the
experiment[20]. The water was changed on alternate days and the concentration of pesticide
was maintained. The pesticide treated fishes (24, 48, 72 and 96 hr) were sacrificed by
decapitation are fixed in fixative. After studying the ovary in situ they were carefully
extirpated, weighed and processed.Serial sagittal sections were cut 5 to 6 µ for further
histological studies during spawning phase. The GSI was also calculated.
RESULT
Control
The ovaries contain ripen eggs with a few immature oocytes. The large oocytes (diameter:
248.53±0.15µ) with large well defined nuclei and distinct nucleoli. The tunica albuginea is
made up of muscular and connective tissue fibers and contain blood capillaries.
Treated
The ovaries of 0.05 ppm of neem oil for 24 hrs, showed decrease in size (GSI: 5.94 ± 0.16 µ).
This was due to presence of large number of V stage oocytes slighty increases the number of
degenerating oocytes. The ovarian growth was inhibited with treatment of neem oil for 24 hrs
in 0.25 ppm. Some of the stages III and IV oocytes exhibited degeneration. The GSI was
comparatively lesser when compared to control (control: 6.98 ± 0.11 µ, treated: 5.54 ± 0.20
µ). Neem oil treatment of 0.5 ppm for 24 hrs, the oogonia had undergone cytoplasmic
clumping and degeneration.
Exposure of fishes to 0.05 ppm of neem oil for 48 hrs, showed atretic follicles, degenerating
oocytes, growing oocytes like III and IV stages were seen to undergo degeneration.The
ovaries of 0.25 ppm of neem oil for the 48 hrs treatment, the oocytes showed a few yolk mass
in the mature oocytes. The nuclear diameter of V and VI stages of oocytes decreases when
compared to that of control. The ovaries of 0.5 ppm of neem oil treatment for 48 hrs,
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oocytes showed maximum enlargement with large yolk oocytes. Some of the stages V and VI
oocytes exhibited follicular degeneration.
Exposure of fishes to 0.05 ppm of neem oil for 72 hrs, the oocytes were seen to undergo
degeneration. Some yolky oocytes undergo atresia. The ooplasm was disorganized and the
zone pellucida had swollen. Fishes exposed to 0.5 ppm of neem oil for 72 hrs, oocytes
showed more number of atretic follicles of stage V and VI oocytes and immature oocytes
were also found.
On treatment with 0.05 ppm of neem oil for 96 hrs, the ooplasm was disorganized and the
zona pellucida had swollen and defected from the original position. The percentage of
oocytes stage VI decreased when compared to control. The ovarian growth was inhibited with
treatment of 0.25 ppm of neem oil for 96 hrs, the oocytes and interstitial tissue were
encapsulated by a connective tissue, consisting of germinal epithelium and tunica albuginea
with blood vessels. There was reduction in the GSI (2.30 ± 0.21 µ). Exposure of fishes to 0.5
ppm of neem oil for 96 hrs, the ovary revealed, large number atretic follicles, further
interfollicular oedema due to shrinkage of oocytes were also appeared and
display an
immense amount of empty follicles and necrosis of nuclei and expelled nuclei . Decreases the
percentage of mature oocytes when compared to control. The results indicated that the
neemoil induced the growing oocytes to undergo further development resulting in early
atresia.
Table-1: The effect of neem oil on oocytes diameter of G. giuris during spawning phase
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Table-2: The effect of neem oil on oocytes percentage of G. giuris during spawning
phase
Graph-1: Effect of neem oil on GSI of gobiid fish G. giuris during spawning phase
DISCUSSION
During the spawning period (September to December) ovaries of Glossogobius giuris contain
ripen eggs with a few immature oocytes. The large oocytes with large well defined nuclei and
distinct nucleoli. Some of the stages III and IV oocytes exhibited degeneration. The GSI was
comparatively lesser, the oogonia had undergone cytoplasmic clumping and degeneration.
[21]
who worked on Tor tor (barbus) also reported that extensive depletion of cyanophilis was
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accompanied by an increase in the number of atretic follicles and oogonial proliferations in
the ovaries. Neem oil has interfered with maturation and growth of the follicles in the ovary
which has also been showed by the microscopic sections of the ovary in which the number of
follicles decreases
[14, 22]
. The inter follicular spaces, as markedly observed in the present
study, has also been observed by many earlier workers in fish exposed to various
pollutants[23,
24,25, 26]
. Ovarian recrudescence has been reported in Channa punctatus and
Clarias batracus after Ion treatment with Hgcl and Emisan
[27, 28]
. Large number ofatretic
follicles, further interfollicular oedema due to shrinkage of oocytes were also appeared and an
display an immense amount of empty follicles and necrosis of nuclei & expelled nuclei and
reduction in GSI, similar changes have also been reported by [29].
SUMMARY AND CONCLUSION
In Glossogobius giuris, ovulated during September to December. Both the gonadosomatic
index (GSI) and the study of ova diameter indicate that G. giuris spawns only once a year
during September to December. The ova diameter and GSI appear to be maximum in
September and is coincident with the peak of spawning activity of the fish. The results
indicate that, neemoil is capable of inhibiting ovarian development and inducing
cytomorphological changes in the oocytes, deformities in nucleoli,interfollicular oedema and
formation of atretic oocytes. A low dose of neem oil brings about a significant reduction in
the Gonado somatic index (GSI).
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