(peganum harmala l.), rhizomes of kuth

Pak. Entomol. Vol. 28, No.1, 2006
EFFECT OF DIFFERENT EXTRACTS OF HARMAL (PEGANUM HARMALA L.), RHIZOMES OF
KUTH (SAUSSUREA LAPPA C. B. CLARKE) AND BALCHAR (VALARIANA OFFICIANALIS L.) ON
THE SETTLING AND GROWTH OF PEACH FRUIT FLY, (BACTROCERA ZONATA SAUNDERS)
Masood Khan Khattak 1 , Mohammad Faisal Shahzad 1 and Ghulam Jilani 2
1. Department of Entomology, Faculty of Agriculture, Gomal University, D.I. Khan.
2. Integrated Pest Management Program, IPEP, NARC, Islamabad.
ABSTRACT
Repellent and growth inhibiting effect of the plants viz. harmal, Kuth and balcher each extracted in petroleum ether,
acetone and ethanol were evaluated at 2%, 1% and 0.5% concentrations against Bactocera zonata Saunder. In a free
choice test in cages, none of the plant at 2% affected the settling of the test insect, however, emergence of progeny adult
was significantly lower in petroleum, ether and acetone extract of Saussurea lappa and ethanol extract of Peganum
harmala. Significantly, lower number of fruit flies settled on guava treated with 1% extract of all the tested plants as
compared to that in the control. Pupal formation and emergence of progeny adult was totally inhibited in 1% petroleum
extract of Saussurea lappa at 0.1%, number of adult settled were significantly lower in petroleum ether and acetone
extract of P. harmala and S. lappa, respectively. However, adult emergence was significantly lower than control in all the
extracts.
INTRODUCTION
Fruit fly, Bactrocera zonata Saund. (Diptera,
Tephritidae) is a major insect pest of fruit and
vegetables not only in Pakistan but the world over as
well. Fruit fly cause 25-50% loss to guava during
summer (Syed, 1970). The different species of fruit
fly cause colossal losses to fruit and vegetables in
tropical, sub tropical and temperate regions of the
world (Batman, 1972). They cause direct loss to fruit
and vegetables and indirect loss by enhancing
expenditure on chemicals which alternately reduce
the market value of the product. The infestation may
reach up to 50% in summer crop of guava in
Pakistan. The peaches, apricot, guava or other fruit
attacked by this pest are malformed, mis-shaped,
undersized and rotted inside. The damage caused is
very heavy and the fruits become unmarketable
(Atwal, 1976). Due to its life cycle, the actual
damage is virtually safe from the insecticide
applications as the eggs are laid in the fruit tissues
and the larvae feed inside on the pulp. Therefore,
control measures are to be directed at the adult flies.
The adults are long lived and quite mobile but their
longevity associated with frequent feeding enables
them exposed to the poison baits and other control
strategies. The enormous loss due to fruit flies in
food production at all level of society demand to
solve this problem with the implementation of wide
management strategies in the area.
Mostly, these insect pests are controlled by synthetic
insecticides for their quick knock down effect,
however, careless and indiscriminate use of these
chemicals leads to a number of well known problems
like contamination of food, soil, ground water, lakes,
rivers, oceans, air with toxic residues which carry side
effects on non target insects and other organisms,
development of resistance in insects against these
pesticides and pest resurgence etc. In addition, many non
lethal and lethal accidents occur due to mishandling of
highly toxic synthetic products. Because of these hazards
of the chemicals, there is growing awareness in developed
and even in developing countries of toxicological and
environmental problems involved in the use of synthetic
pesticides. This awareness has lead to a steadily increasing
movement toward more environmental oriented,
sustainable agriculture with low or no input of toxic
synthetic material and other agriculture chemicals in an
attempt to preserve and protect the environment and as
well as human health.
Plant kingdom is by far the most efficient factory of
compounds. Plants synthesize countless products which
can partially be considered as weapon to defend plants
against pests and diseases. Plants derivatives are less toxic
or non toxic to mammals and other vertebrates and
invertebrates. Plant products have several uses in insect
control. Some act as attractants like geranil and eugenol
while some act as repellent, deterrent and antiovipositional
like neem oil and cotton seed oil etc. The
primary role, however, is insect toxicants (Hashimi, 2001).
Out of 2600 plant species in the insect pests control
potential, pyrethrum, nicotine, rotenone, ryania, sabadilla
and neem have been registered for significant use as
insecticides (Dhaliwal and Heinrich, 1998).
Different formulations prepared from neem and other
plants like, Pongamina, datura, thevatia etc. have proved
promising in checking insect pest population. The most
important aspect in the use of plant products against insect
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Pak. Entomol. Vol. 28, No.1, 2006
pest is that these naturally occuring materials are non
toxic to man and other animals and are environment
friendly.
METHODS AND MATERIALS
Settling and Growth Regulating Effects of Certain
Plants Extracts
To test the repellent/deterrent and growth inhibiting
effects of certain plants extracts against peach fruit
fly, Bactrocera zonata (Saunder), studies were
carried out at Insect Pest Management Program
(IPMP), Institute of Plant and Environmental
Protection, National Agriculture Research Center,
Islamabad.
Seed of Harmal (Peganum harmala L.), Rhizomes of
kuth (Saussurea lappa C.B.Clarke) and Balchar
(Valariana officianalis L.) used in these studies were
received from the Agervedec shop in Rawalpindi.
Peach fruit fly (Bactrocera zonata) used in these
studies were obtained from the culture maintained in
the laboratory with 27±1C 0 , 60±5% relative humidity
and 14:10 light and dark. Fruit flies were reared on
artificial diet in fruit fly rearing cage (18 x 16 x 16)
having muslin cloth on the two opposite sides. The
other two sides top and bottom were made of
transparent glass sheet. On alternate day, fresh fruits
were offered to adult flies for oviposition. After
infestation, these fruits were placed in glass jars
having sand at the base for pupation. To allow
aeration the jars were covered with muslin cloth. The
pupae were separated from the sand with 20m sieve.
Pupae were placed in rearing cage with cotton soaked
in water to prevent dessication. After emergence of
flies, pertinacious diet was provided on daily basis to
the flies. For water requirements water soaked cotton
was placed in Petri-dish inside the rearing cage.
Artificial diet for fruit flies
Ingredient
Bananas
Egg yolk
Honey
Lederplex vitamin B. Complex syrup
Yeast
Sugar
quantity
2 No
6 No
4 table spoon
2 table spoon
1 table spoon
8 table spoon
The ingredients were mixed in a blender to make a
thick syrup solution. For further use the diet is kept in
refrigerator. Products of all the tested three plants
were separated, ground to fine powder of 60 mesh.
The plant powder were separated, extracted with
petroleum ether, acetone and ethanol in Soxhlet
extraction apparatus, concentrated in rotary
evaporator and finally made solvent free in a vacuum
dessicator. Since plant extracts are insoluble in water
therefore, a small quantity of detergent (Surfactant) was
added in distilled water.
To see the effect of test plants extracts on the settling of
fruit flies. All the plant extracts were applied at 2%, 1%
and 0.5% to the fruits separately by dipping method. The
treated and untreated fruits were placed randomly in the
rearing cages. Fifteen day old 40 female fruit flies were
released in the cage. Each cage providing free choice was
replicated three times. The number of fruit flies adults
settled on treated and untreated fruits were counted after
every two hours for two days. In this way the repellency/
deterrence was counted. After 48 hours the exposed treated
and untreated fruit to fruit flies were removed and placed
in separate plastic jars with sand in it for fruit flies
pupation. The jars were covered with muslin cloth and
properly labeled. Each treatment was replicated three
times. After 20 days the sand was sieved with 20 mesh
sieve. The pupae were separated and counted and data
recorded for each treatment. The pupae were sticked to
glass vials. The vials were covered with muslin cloth and
labeled. Cotton soaked in water were placed at the top of
vials to provide moisture needed to prevent pupal
dessication. The emergence of fruit fly adults were
recorded for each treatment.
RESULTS
Petroleum ether, acetone and ethanol extract of P.
harmala, V. officianalis, and S. lappa when applied at 2%
to guava fruits did not significantly affected the settling or
non preference response of the fruit flies (Table-1); as the
mean number of flies settled on fruit treated with different
extracts of plants at 2% concentration were statistically
similar to those settled on the untreated guava fruit.
However, as compared to 0.6867 adults in the control, the
lowest number of fruit flies adults settled on guava fruits
treated with 2% acetone extract of V. officianalis. This was
followed by 0.2633 and 0.3733 flies in 2% petroleum ether
extract of S. lappa and 0.3967 adult ethanol extract of V.
officianalis.
When the same exposed fruits were placed separately for
pupation the number of pupae in some treatments were
significantly lower than those in the control (Table-2).
Minimum number (2.667) pupae were counted each from
the fruits treated with ethanol extract of P. harmala and
petroleum ether of S. lappa followed by 3.00 from those
treated with acetone extract of S. lappa. These number
were statistically lower than 11.33 pupae in the control.
The fruits treated with petroleum ether extract of P.
harmala had 7.00 pupae, which were still significantly
lower than those in the control but not from the remaining
treatments. (Table-2). The trend of adult emergence was
similar to that of pupae. Minimum and significantly lower
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Pak. Entomol. Vol. 28, No.1, 2006
number of adult emerged at the same treatment in
which pupae were minimum and significantly lower
than other treatments and control.
Settling response by B. zonata on guava fruits treated
with 0.5% concentration of petroleum ether, acetone
and ethanol extracts of P. harmala, V. officianalis
and S. lappa in a free choice test is given in Table-3.
It reveled that flies settled on fruits treated with 0.5%
Petroleum ether extract of P. harmala and Petroleum
ether extracts. In these treatments only 0.1300 and
0.0767 flies settled on the treated fruits which were
statistically lower than the 0.8467 flies settled on the
untreated fruits. In other treatment, though, lower
number of flies settled on the treated fruits were not
statistically different to that on the untreated one.
The number of pupae recovered in all these
treatments were significantly lower than the pupae
obtained in the control except for the pupae
recovered from fruits treated with acetone extract of
P. harmala. The lowest number (1.667) pupae were
obtained from the fruits treated with acetone extract
of S. lappa, however, were statistically similar to the
pupae recovered in the other treatments except
control. Almost, similar trend was observed in the
adult emergence. Significantly lower number of
adults emergence in all treatments as compared to
those in the control.
The above results showed that there was no
correlation between the number of adult flies settled
and progeny recovered. This phenomenon clearly
indicate that some of the treatments affected both
settling as well as egg laying response while others,
were partially effective. Extracts of the tested plants,
when applied at 1% to guava fruits, significantly
reduced the settling of fruit fly on treated fruits as
compared to the untreated ones. However, fruit fly
adults settled on the treated fruits with different
extracts were statistically similar to each other
(Table-3). Petroleum ether extract of V. officianalis
with 0.2233 adults ranked first in showing non
preference by test insect followed by 0.2467, 0.2867
and 0.3500 adults on guava fruits treated with 1%
ethanol extract of P. harmala, acetone extracts of P.
harmala, ethanol extract of V. officianalis and
petroleum ether extract of P. harmala, respectively.
When the fruits used in this study were keep
separately in jars with sand at their bottom,
statistically similar pupae were recovered in all
treatments as compared to those in the control except
of S. lappa where no pupae were recovered. Hence,
when there were no pupae, therefore, were no adult
recovery, though the number of flies settled on these
fruits were statistically similar to those on the other
treatments.
Table-1. Mean number of Bactrocera zonata settled and progeny recovered
from untreated Guava fruits and those treated with 2% concentration of
petroleum ether, acetone, and ethanol extract of different plants in a free choice
test
Plant
Mean Number of
Solvent 2% Adu. Settled Pupae Recovered Adu. Emerged
Peganum harmala
Petroleum ether 0.5533 a 7.000 b 7.000 bc
Acetone 0.4433 a 8.000 ab 7.000 bc
Ethanol 0.4433 a 2.667 c 1.333 d
Valariana officianalis
Petroleum ether 0.5500 a 11.33 a 9.667 ab
Acetone 0.2600 a 8.33 ab 6.333 c
Ethanol 0.3967 a 10.33 ab 10.00 a
Sassurea lappa
Petroleum ether 0.2633 a 2.667 c 2.333 d
Acetone 0.3733 a 3.000 c 2.000 d
Ethanol 0.6633 a 9.000 ab 8.000 abc
Control 0.6867 a 11.33 a 7.667a
Each value is a mean of three replications, 10 pairs per replication. Mean
values among the columns followed by the same letters are not significantly
different at α = 0.05, having LSD value = 0.4912 for settling, LSD value = 3.695
for pupae, & LSD value = 2.639 for adult
Table-2. Mean number of Bactrocera zonata settled and progeny recovered
from untreated Guava fruits and those treated with 1% concentration of
petroleum ether, acetone, and ethanol extract of different plants in a free choice
test
Plant
Mean Number of
Solvent 2% Adu. Settled Pupae Recovered Adu. Emerged
Peganum harmala
Petroleum ether 0.3500 b 3.667 abc 3.667 ab
Acetone 0.2867 b 3.667 abc 2.333 ab
Ethanol 0.2467 b 6.670 bc 1.000 ab
Valariana officianalis
Petroleum ether 0.2233 b 2.667 abc 2.333 ab
Acetone 0.5167 b 8.000 abc 6.667 ab
Ethanol 0.2867 b 4.000 abc 3.667 ab
Sassurea lappa
Petroleum ether 0.4367 b 0.000 c 0.0000 b
Acetone 0.4333 b 3.667 abc 3.667 ab
Ethanol 0.4333 b 10.33 a 8.000 a
Control 1.0200 a 8.333 ab 7.667 a
Each value is a mean of three replications, 10 pairs per replication. Mean
values among the columns followed by the same letters are not significantly
different at α = 0.05, having LSD value =0.3431 for settling, LSD value = 7.041
for pupae, and LSD value = 6.127 for adult
Table-3. Mean number of Bactrocera zonata settled and progeny recovered
from untreated Guava fruits and those treated with 0.5% concentration of
petroleum ether, acetone, and ethanol extract of different plants in a free choice
test
Plant
Mean Number of
Solvent 2% Adu. Settled Pupae Recovered Adu. Emerged
Peganum harmala
Petroleum ether 0.1300 b 4.667 b 3.333 b
Acetone 0.4900 ab 12.67 ab 10.33 b
Ethanol 0.4633 ab 8.333 b 7.333 b
Valariana officianalis
Petroleum ether 0.433 ab 3.000 b 2.000 b
Acetone 0.3567 ab 7.000 b 6.333 b
Ethanol 0.3133 ab 3.333 b 2.333 b
Sassurea lappa
Petroleum ether 0.5133 ab 3.000 b 2.000 b
Acetone 0.1767 b 1.667 b 0.000 b
Ethanol 0.5567 ab 5.000 b 3.333 b
Control 0.8467 a 23.00 a 22.00 a
Each value is a mean of three replications, 10 pairs per replication. Mean
values among the columns followed by the same letters are not significantly
different at α = 0.05, having LSD value =0.5287 for settling, LSD value = 12.01
for pupae, and LSD value = 9.910 for adult
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Pak. Entomol. Vol. 28, No.1, 2006
DISCUSSION
To reduce losses by fruit flies, farmers mostly use
synthetic insecticides which cause serious social and
environmental repercussions. Male Inhalation
Technique (MIT), Bait Application Technique
(BAT), sanitation of orchards are very effective and
friendly management measures but are not adapted
by the majority of farmers, therefore, it is felt that
fruit flies can be effectively controlled only through
integrated pest management using environmental
friendly materials to replace insecticides.
Among various plant extracts applied to guava fruit
in the laboratory studies, Valariana officianalis
extract in acetone and petroleum ether significantly
repelled the fruit flies from the treated fruit.
Petroleum ether and acetone extracts of Saussurea
lappa although, was not effective in reducing the
settling of flies on treated fruit, it significantly
affected the pupal formation and adult emergence.
This indicated that Saussurea lappa had suppressed
egg laying. Chander (1999), Harish (2000) found that
these plants oil significantly repelled major insect
pests of stored grain and growth of certain stored
grain insect pests was impaired . Neem seed kernel
extracts worked as oviposition deterrent to
Bactrocera curcurbitae and B. dorsalis on pumpkin
and guava, respectively. The acetone extract of deoiled
kernel powder significantly deterred oviposition
by both species at 0.5% and 1.25% (Shivendra et al.,
1998). Renden et al. (1998) investigated a strong
negative effect of neem based insecticides on the
oviposition, egg development and survival of the
adults.
In the present studies, the tested plant derivatives
significantly affected the growth of fruit flies.
Similar, results were obtained by Hassan (1998),
according to him, neem seed kernel extract (NSKE)
negatively affected the 1 st and 2 nd instar larvae of the
Queensland fruit fly on persimmon. A high
proportion of adults emerging from the treated pupae
showed wing deformation preventing them from
flying. Shakunthala et al. (2001) demonstrated
remarkable changes in the size and morphology of
the reproductive organs of adult flies when Acorus
calamus extract were used against B. cucurbitae.
10% ether extract of Peganum harmala significantly
increased adult mortality, retarded rate of
development, reduced adult emergence of
Callosobruchus chinensis.
Some botanicals are also used as attractants for fruit
flies. Kardinam (1999) found that Melaleuca
bracteala oil containing methyl eugenol was effective in
trapping fruit flies. Shivendra et al. (2001) tested five
fractions of Acorus calamus oil against Bactrocera
cucurbitae and B. dorsalis. Fl fractions were attractive to
male Bactrocera dorsalis. FII (acoragermacrone) was
more attractive to female flies.
LITERATURE CITED
Atwal, A.S., 1976. Agricultural pests of India & Southeast
Asia, Kalyani Publishers, New Delhi.
Bateman, M.A., 1972. The ecology of fruit flies, Ann. Rev.
Ent., 17: 493-518.
Chander, H., 1999. Laboratory evaluation of plant extracts as
repellent to the rust flour beetle, Tribolium
castaneum on jute fabric, Internat. Pest Contr., 41
(1): 18-20.
Dhaliwal, G.S. and R.A. Heinrich, 1998. Critical Issues in
insect management. Common wealth publishers,
New Delhi.
Harish, C., 2000. Repellency of different plant extracts and
commercial formulations used as prophylactic spray
to protect bagged grain against Tribolium
castaneum, J. Food. Sci., and Techa. Mysore. 6(37):
582-585.
Hashmi, A.A., 2001. Integrated pest management in the 21 st
century, PARC publication, Islamabad.
Hassan, E., 1998. Insecticidal toxicity of neem seed kernel
extract (NSKE) on Bactrocera tryoni (Frogg.)
(Diptera: Tephritidae) and repellency on persimmon
fruit. Zeitschrift-fur-Pflanzenkrankheiten-und-
Pflanzenschutz. 105 (4): 411-416.
Kardinan, A., 1999. Prospect of Melaleuca bracteata leaf oil
as fruit fly Bactrocera dorsalis control in Indonesia,
Journal Penelitian and Pengembangan Pertanian.,
18(1): 10-17.
Renden, E.J., B.D. Roitberg and E.J. Randen, 1998. Effect of
a neem Azadirachta india based insecticide on
oviposition deterrence, survival, behavior and
reproduction of adult western cherry fruit fly
(Diptera: Tephritidae), J. Eco. Ent., 91(1): 123-130.
Shakunthala, N. and J. Thomas, 2001. Ovipositior deterrence
of Acorus calamus L. on melon flu, Bactrocera
cucurbitae, Insect Environm., 39(2): 145-148.
Shivendra, S. and R.P. Singh, 1998. Neem Azadirachta indica
seed kernel extracts and azadirachtin as oviposition
deterrents against the melon fly Bactrocera
cucurbitae and the oriental fruit fly Bactrocera
dorsalis. Phytoparasitica, 26(3): 191-197.
Shivendra, S., S.S. Sehgal and S. Singh, 2001. Investigations
on constituents of Acorus calamus root oil as
attractants to melonfly, Bactrocera cucurbitae and
oriental fruit fly, Bactrocera dorsalis. Indian J. Ent.,
63(3): 340-344.
Syed, R.A., 1970. Studies on the trypetids and their natural
enemies in West Pakistan. Dacus spp. of lesser
importance, Pak. J. Zool., 2: 17-24.
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