Journal of Mosquito Research 2015, Vol.5, No.10, 1-7
5
Table 7 Completely randomized three ways ANOVA analyses using instars (I) of
Cx. quinquefasciatus
, hours (H), and Concentrations
of ethyl acetate root extract of
Annona reticulata
(C) as three independent parameter
Source of variation
Sum of squares (SS)
Degree of freedom (df)
Mean of squares (MS) F value
p-level
Instars (I)
200.25
3
66.75
457.71
0
Time (H)
59.76
2
29.88
204.90
0
Conc.(C)
107.86
3
35.95
246.54
0
I×H
12.29
6
2.05
14.05
0
I×C
26.08
9
2.90
19.87
0
H×C
34.51
6
5.75
39.44
0
I×H×C
5.21
18
0.29
1.98
0.0177
Within groups
459.97
96
0.15
-
-
Total
0.95
143
3.22
-
-
of above mentioned works with the result of present
study, our work is better than their works. So it may
be concluded that crude and different solvent root
extracts of
Annona reticulata
can be used effectively
to control
Cx. quinquefasciatus
mosquito larvae in
their breeding places because different extracts work
at very low concentrations. Further research is needed
to know the name (s) and chemical structure (s) of
actual compound (s) which is responsible for larvicidal
activity.
3 Materials and Methods
3.1 Collection of plant parts
After proper identification of the
A. reticulata
plant,
roots of near about one to two years aged plants were
collected during September and October, 2014 from
Burdwan town, West Bengal, India (23
◦
16
ꞌ
N, 87
◦
54
ꞌ
E)
and the voucher specimen (No.GCZSM-4) of the plant
was kept as herbarium to Mosquito, Microbiology and
Nanotechnology Research Units
,
Parasitology Laboratory,
Department of Zoology, The University of Burdwan,
West Bengal, India. After collection, roots were
washed with tap water and then rinsed with distilled
water and dried on paper towel.
3.2 Preparation of crude extract
Washed roots were cut into small pieces and crushed
with mechanical grinder and juice was filtered by
Whatman No. 1 filter paper and the filtrate served as
stock crude test solution. From stock crude test solution
required graded concentrations (0.01- 0.05%) of crude
extract was prepared through dilution with tap water.
3.3 Preparation of solvent extracts
Washed roots were cut into small pieces and dried for
15-18 days in shade with gentle blowing of air. Dried
small pieces of roots (150 g) were packed into the
thimble of Soxhlet apparatus and passed petroleum
ether, hexane, and ethyl acetate solvents (Volume of
each solvent 1500 ml). For each solvent extraction,
fresh root samples were used and the extraction period
of different solvents were 72 h. Pooled extracts were
concentrated by rotary evaporator and semisolid
extracts of different solvents were obtained and stored
at refrigerator at 4
℃
for further use. 0.05 g semi solid
extract of different solvents were dissolved initially on
5 ml of ethanol and then added 95 ml of distilled water
to obtain 100 ml of stock test solution of different
extracts. So stock solution of different solvent extracts
were prepared on 5% ethanol. From stock test solutions,
1, 2, 4, and 8 ppm concentrations of petroleum ether,
hexane, and ethyl acetate solvent root extracts of the
plant were prepared through dilution with tap water
which were used for larvicidal bioassay experiments.
3.4 Test mosquito
Cx. quinquefasciatus
were collected from cemented
drains of Burdwan town and identified properly and
the larvae were kept in plastic tray with tap water.
Larvae were well maintained in the laboratory at 27±
2
℃
temparature and 85% relative humidity under 14:10
light and dark cycles in a day. The larvae were fed with
powdered mixtutre of dog biscuits and dried Brewer’s
yeast powdered (Ratio 3:1). Pupae were transferred from
plastic tray to a plastic bowl containing tap water and
bowl was kept in mosquito cage (30×30×30 cm
3
) where
adults were emerged. Adults were provided with glucose
solution (10%) in a plastic bowl with a cotton wick.
Adults were provided blood meal from restrained pigeon
on day five. Plastic bowl with 100 ml of tap water were
kept in the cage for oviposition. Next generation larvae
were used for bioassay.
