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r EFFICACY OF CERTAIN TOXIC COMPOUNDS EXPOSED TO
AGRICULTURALLY IMPORTANT INSECT PEST IN RELATION TO SOME ABIOTIC FACTORS
DISSERTATION
SUBMJTTIEDIN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE A WARD OF THE DEGREE OF
f a s t e r nf ptfilnsnplig
BY
ASAF ALI MURTAZA
Under the supervision of
DR. KHOWAJA JAMAL
1 DEPARTMENT OF ZOOLOGY FACULTY OF LIFE SCIENCES
ALIGARH MUSLIM UNIVERSITY ALIGARH (INDIA)
2011
DS4184
f e d ^^ '•^'••'-
' External: 2700920/21-3430
iBtcrnal: 3430
DEPARTMENT OF ZOOLOGY ALIGARH MUSLIM UNIVERSITY
ALIGARH-202002 Sections: INDIA D.No ../ZD
1. Entomology Dated 2. Fishery Science & Aquaculture 3. Genetics 4. Nematology 5. Parasitology
ST. Jissistant (Profassor-Cum-^useum Curator
l^w is to certify that the research wor^presentecC in the dissertation
entitled 'efficacy of certain toxic compounds exposed to agricutturaSy
important insect pest in relation to some aSiotic factors' carried out 6y
Mr. AsafjiR 9iurtaza under my supervision. The dissertation is an ori^inaf
contriSution of the researcher and adds suSstantiaCCy to the exiting ^owledge
in the discipfine. He is permitted to su6m.it it for the partiaCfuffiUment of the
degree of Master of (Philbsoplty in Zoology, at Migarh MusCim Vniversity,
AGgarh, India. The dissertation is fit for suSmission to the ej^aminers for
evaCuation. »
It is further certified that Mr. JisafM Murtaza has fuCfiOed the
prescriSed conditions of duration and nature given in the Ordinances of the
ACigarh Muslim Vniversity, Migarh. " '
(Dr, Khowaja Jamal) Sr. Assistant Professor-Cum-Museum Curator
Supervisor
AcknomUinmnent
Tirst andforemost, I -woufdCi^ to pay my oBeisance and gratitude to "JtHah
JiCmigHty" thefaitd in wHom fieCpedme aCways.
It gives me immense pleasure to express my heartfeCt gratitude, indebtness
and dumSa regards to my esteemed teacHer and supervisor (Dr. %lwwaja JamaC
JLssistant (Professor, (Department of Zoology, JiMV, JiGgarfi. It is indeed a matter
of great pride and experience -witfiout comparison to Rave had the privilege of
•wor^ng umler his ceaseless and untiring guidance. The present wor^has cuCminated
due to his expert supervision, meticulous guidance and solves each and every proSlem
that I cropped up during the completion of this wor^ His immense knowledge and
constant help irrespective of how Busy his schedule was proved instrumental in
completion of this endeavor.
I would li^ to pay my sincere regards to (Prof. Irfan Ahmed, Chairman
(Department of Zoology, for giving me opportunity to wor^ and providing me laS
and other facilities required during my research wor^
I tender my heartfelt than^ to all my teachers especially
(Dr. Ayesha Qamar Asst. (Professor, (Department of Zoology and
(Dr. (Bachchan Ad "Kfum, for their constant help untiring encouragement, moral
support, valuaSle suggestions, inspiration and profound encouragement for
Betterment, which have Been a driving force for me to perform this wor^
I would li^ to shower my special than^ to (Dr. fHd Sadre ACam Ta^ri,
<Dr Mohd <Fannan 'Kfian, (Dr. Shamim Ahmed, (Dr. Shah Mohammad Haris, the
Budding scientists in entomology, who have Been a source of inspiration for me, too^
painsta^ng efforts during my wor^ fuelled me with his viBrant, constructive and
informative ideas and assisted me in giving the solid shape to my research wor^
m
/ tfian^ffy express my deep sense of gratitude to JAUah for giving me nice
fattier and Motfier, wHo raised me witfi a Cove of science and supported me in ad my
pursuits. What I am today is Because of my parents, their prayers, Cove, sacrifices,
sincere endeavors and sustained efforts tHat enaBCedme to acquire tfiis ^rurwCedge.
9Ay fieartfeCtfeeCings of gratitude to my grandfatRer, Cote grandmother, (Bmtfiers,
sister and ad the family numbers whose prayer and support are aCways with me and
have provided me. aCfthefaciCities whenever needed.
I aCso ta^ this opportunity to my speciaC than^ to my friends
Syed !Hassan MehntR, Jdrdesh. lOimar Sharma, SaBiur ^fiAman ^arooqi,
^ofuf Hamayun, ^ja CftoudRary, Mofid iBraAim, Q?an^j gwari, (Dr. !Mohd
Tauseef, fMofid^fi "Kfian, Mofid Inran, fW<£ Siraj Maneer, <^fyaz Ahmed Mir,
Iram "Kfian, TaSassum <Stfhmat and Tiza %fian for their support at every stage
whenever needed. My time at JACigarh MusRm University, yiCigarh was made
enjoyahCe in Carge part due to many friends that Became a part of my Cife.
Last But not the Ceast, I express my speciaCthan^ to Mrs. Tfasreen iHiissain,
Seminar Librarian, and aCf non-teaching staff of my department especially
%fiaCiq (Bhai and Zaheer (Bhai, who extended ad sorts of heCp and support to me
throughout the compCetion of this wor^
I am deepCy indeBudto Mr. A-% Azad, Mr. AhdufMannan (Munna), Mr. ShoaiS
Ahmad, Mr. Md SuhaiC dC Mr. Shafaqat-MemBers ofAH-In-One Compmmters,
AMI)., ACigarh for composing and (Designing this dissertation.
(Before concCuding, I douBt that I have forgotten to mention and
ac^owCedge many persons who heCp me in one way or the other.
(AsafAS OHurtaza)
m
CONTENTS
INTRODUCTION 1-7
REVIEW OF LITERATURE 8-20
MATERIAL AND METHODS 21 -26
RESULTS AND OBSERVATION 27-46
TABLES 47-54
GRAPHS 55-72
PHOTOGRAPHS 73-74
DISCUSSION 75-96
REFERENCES 97-123
INTRODUCTION
Cotton is an important agricultural crop commonly known as
white gold. It provides major employment in agriculture and textile
industry. The world cotton production estimated at 25.9 million
tonnes in 2007-08, 3% lower than the previous season due to
decline in area under cotton cultivation (Anonymous, 2008).
Whereas, India produced 5.3m tonnes with 11% increase
compared to the previous season because of an increase in cotton
production area by 4% touching 9.58 million ha. However, the
average productivity of cotton in India (589lint/ha) is low as
compared to other cotton producing countries (607 to 1390lint/ha).
There are two types of cotton grown in India, Desi cotton
{Gossipium arboretum L.) and American cotton (G. hirsutum L.).
The Desi cotton is grown in mountain and high rainfall areas. The
American cotton is generally grown where there is some source of
irrigation. Amongst the various constraints in cotton cultivation, the
losses caused by insect pests are of major importance. In India,
160 species of insect and non-insect pests have been reported to
damage cotton crop (Agarwal, 1978). Of these, 24 phytophagous
insect species have attained the pest status, out of which nine are
key pests, in different zones of the country. In the north zone
(Punjab), sucking pests and bollworms cause 11.6 and 49.5% loss
Introducticm
in seed cotton yield respectively and both together caused 52.1%
loss (Dhawan eta!., 1998).
The cotton strainer, Dysdercus koenigii Fabr., is an important
pest of cotton (Gossipium sp) in Punjab and Uttar Pardesh and
widely distributed throughout India. Due to feeding on half opened
cotton bolls and seeds, the oil content and seed viability is reduced
and the yellow excreta of the insect imparts a dirty colour and
spoils the cotton lint. On potted plants, the eggs were laid in cracks
and crevices of soil and covered by loose soil or dried leaves. The
average egg laying capacity of female is 288 (Chatterjeet et al.,
1985). Eggs are oval, smooth, creamy yellow in color with an
average incubation period of 6.18 days. The period of the first,
second, third, fourth and fifth instars range from 3 to 4, 3 to 4, 3 to
5, 6 to 8 and 12 to 16 days, respectively. The adults live for 9 to 25
days and the complete life cycle extends from 51 to 61 days. The
sex ratio of male to female was 50.50 to 48.50. This bug is
prevalent throughout the year and also feeds on okra, hollyhock,
eggplant and other malvaceous plants (Kamble, 1971). The feeding
habits of nymphs and adults were similar except for their
gregariousness during feeding, when food was scarce nymphs and
adults were commonly cannibalistic.
- 2 -
Iianktuction
The chemical control method of the insects is very effective
and rapid in curative action. Although the method is, in general,
ecologically unsound and has many serious limitations like
insecticide resistance, pest resurgence and pest replacement
which can occur following repeated application of these
compounds (Pedigo, 1989), it has been considered as the most
important and powerful tool in controlling insect pests both in the
field and under storage conditions for the past. If we compared with
the alternatives, the benefits of insecticides probably outweigh the
risks. But the goal of insecticide use to obtain maximum benefits
with minimum risks has not yet been achieved in most situations.
This may be achieved by proper use of pesticides and dose
selection,, to follow the rules of pesticide application. A
developmental program for chemical pesticides should involve a
thorough examination of the physical, biological and environmental
factors that can affect the pesticide toxicity. Although there is also a
need to replace dependence on environmentally disruptive
chemicals, it seems likely that the further agriculture production
system will require some new generation insecticides. The
chemical groups, mostly in use today, are organophosphates,
carbamates and synthetic pyrethroids which have broad-spectrum
activity. However, new generation pesticides like imidacloprid,
Introduction
spinosad, fipronyl are much selective, less disruptive and more
environmental benign and having new novel mode of actions,
hence likely to be more compatible with IPM.
Imidacloprid is a relatively new, systemic insecticide
chemically related to the tobacco toxin nicotine. It was first
registered for use as a pesticide in the U.S in 1994. It moves
through the plant from the place where it was applied and kills
insects when they feed. Like nicotine it acts on the nervous system.
Because of the molecular shape, size and charge, it fits into
receptor nnolecules in the nervous system that normally receive the
neurotransmitter acetylcholine. Acetylcholine transmits nerve
impulses from one nerve cell to another or from nerve cell to some
other tissues like muscles or glands. Nervous system of insects as
well as mammals has acetycholine receptors; most of the sensitive
receptors are in the nervous system of insects, but in nerves
associated with muscles in mammals (Ware, 2000). However,
insect acetylcholine receptors are more sensitive to imidacloprid
than are mammalian receptors (Zwart etal. 1994). Imidacloprid has
high affinity to insect nicotinic acetylcholine receptors (nAchR). It
interacts with nAchR, resulting in excitation and paralysis followed
by death of the insect. Their selectivity results from a higher affinity
to the insect nAchR as compared to that of vertebrates, in contrast
Intfoductum
to the original nicotine compounds (Tomizawa et al., 1995). Hence,
it can be suggested that imidacloprid and related compounds be
called as nicotinoids (Yamento et al., 1995). The nicotinoids act
especially on sucking pests and have mild or no effect on
parasitoids and predators and as such fit well in various IPM
programs.
Spinosad (Dow Agrosciences), a mixture of spinosyns A and
D that are tetracyclic macrocyclic compounds produced by an
actinomycete Saccharopolyspora spinosa. Mertz and Yao isolated
from a jamican soil sample (sparks et al., 1998). As these products
are created biosynthetically during fermentation of S. spinosa,
spinosad has been classified as a bio insecticide (Cropping and
Menn, 2000). It is primarily a stomach poision with some contact
activity and is particularly active against Lepidoptera and Diptera. It
is a neurotoxin with a novel mode of action involving the nicotinic
acetylcholine receptor and apparently the GABA receptors as well
(Salgado, 1997, 1998). Exposure of spinosad results in cessation
of feeding followed by paralysis and death. Conventional toxicity
tests indicate that spinosad has virtually no toxicity to birds and
mammals, with contact LC50 value of 200ppm. Spinosad has also
been reported to be practically non-toxic to natural enemies of
insects(Schoonover and Larson, 1995; Bretefa/., 1997).
- 5 -
Introduction
Furthermore, insecticide use is generally the preferred
strategy for its high efficacy during a pest outbreak. In some
environments such as greenhouses, storehouses and quarantine
facilities, with their relatively closed conditions, high temperatures
can be used to control insect pests. In previous studies
(Alexandresu, 1986; Turnbull and Harris, 1986; Horn, 1988; Morytz,
et ai, 1997; Cho et ai, 1999; Rahman et al. 1999) the effects of
temperature on toxicity of some insecticides were investigated.
Generally, the toxicity of chlonnated hydrocarbons,
organophosphates and carbamate compounds increases with
temperature i. e. exhibit positive temperature coefficient, while
toxicity of some pyrethroids has negatively correlated with
increasing temperature (Hinks, 1985; Grafius, 1986; Thaung and
Collins 1986; Horn 1988; Johnson 1990; Arthur 1999; Wadleigh et
al., 1991). The degradation rates of most organophosphate
insecticides generally increase rapidily with increase in temperature
and either grain moisture content or relative humidity (Snelson,
1987; Arthur et al 1992). Although pyrethroid insecticides are
generally' more stable than organophosphates, increased
temperatures will also enhance pyrethroid degradation (Noble et al.
1982). The realized efficacy of an insecticide is a function of the
inter-relationship between vaporization and cuticular penetration.
Introduction
Residual activity of an insecticide is usually correlated with
temperature. Generally, an insecticide will remain active for longer
periods when temperatures are lower, due mostly to reduced
vaporization (Felsot 1989). The influence of temperature on the
efficacy of insecticides in the field has usually agreed with results of
bioassays in the laboratory (Ayad and Alyousef, 1984; Chandler, et
al., 1991) but very few studies have dealt with the association
between insecticides and abiotic factors. Therefore, the present
study aim's to investigate the effects of temperature, photoperiod
and constant relative humidity on the toxicity of different classes of
insecticides to Dysdercus koenigii (Fabr.) in laboratory.
- 7 -
REVIEW OF LITERATURE
There are certain factors that affect the growth and reproduction
of insects. These factors are endogenous as well as exogenous.
Several author studied the effect of various exogenous factors such as
temperature, humidity, light, food, chemicals etc on various
physiological processes including mortality, longevity, fecundity, fertility,
and malformation.
Abdullah et al., (2002) tested the efficacy of two insecticides
imidacloprid and temik against whitefly and jassid and reported that
both insecticides were good in controlling the jassid and whitefly
population.
Wang et al., (2008) determined the efficacy of sub lethal
concentrations (LC50) of six insecticides (imidacloprid, rotenone,
fanvalerate, abamectin, pericarb and azadirachtin) on fecundity and
wing dimorphism of green peach aphid Mysius persicae under
laboratory and green house conditions separately. In laboratory
condition, after being exposed to sub lethal doses of imidacloprid and
fanvalerate, the proportion of alate progeny in aphid progeny were
significantly higher than that of the control while in green house, the
percentages of alate offsprings from the mother aphid treated with
imidacloprid and fanvalerate and abamectin increased pronouncedly
compared with controls. Mortality rates of offsprings in the nymphal
^^fviewofLiumture
stages from adults treated with insecticide revealed no significant
differences between laboratory and green houses.
Richman et ai, (1999) investigated the toxicity of imidacloprid to
cat fleas on glass at 20, 26, 30, and 35°C and found that it is the most
toxic to adult cat fleas at 35°C and to larvae at 20°C. Further he
observed that piperonyl butoxide (PBO), a synergist, increased the
relative potency of imidacloprid (1:5 imidacloprid:PBO) 16-fold at 26°C
against adults, but had no effect at 35°C. No synergism occurred in
larvae at 20°C, but addition of PBO (1:5 imidacloprid: PBO) doubled
toxicity at 26°C. PBO (1:5 imidacloprid: PBO) could possibly be used to
synergize imidacloprid premise treatments (20-30°C), but it is not likely
to be effective in pet treatments because no synergism occurred in adult
fleas at 35°C (average fur temperature of tested cats and dog).
De Cock et ai, (1996) investigated the susceptibility of the
predatory bug Podisus maculiventris (Say) to imidacloprid in the
laboratory through direct topical exposure, residual contact and
ingestion and observed that imidacloprid toxicity decreases in order of
topical exposure> ingestion > residual contact.
Nishi and Takashashi (2002) reported that the pre-oviposition as
well as egg incubation period of Amphibolus venator decreased with
increasing temperature, whereas, the average number of eggs laid per
-9-
^fviewofLiUmtttn
ovipositing female increased with increasing temperature, however, the
hatchability remain unaffected.
Ali and Qureshi (1982) observed that virgin females of Dysdercus
cingulatus lay no eggs, however the topical application of the juvenile
hormone analogue methoprene (Z15) on the thorax resulted in some
oviposition by virgins. Further he found that the number of eggs laid
increased with increasing doses, but it was never as great as the
number laid by mated females. Treatment of both mated and newly
emerged females with increasing concentrations of methoprene
resulted \n progressive reductions both in numbers of eggs laid and in
the time required for the completion of vitellogenesis.
Yasmeen et ai, (1984) assesed the iox\c\iy of juvenile hormone
analogue methoprene against 5' instar nymphs of Dysdercus koenigii in
the laboratory and reported that the infection of LD50 dose i.e.
45|jg/nymph caused abnormalities in the 5th-instar, whereas, the topical
application of LD50 i.e. 28|jg/nymph prolonged the mstar duration. Chitin
synthesis and moulting were also inhibited.
Ansari et al., (2008) studied the total effect of imidacloprid on
development of Plutella xylostella and reported that feeding of 3" instar
larvae on cauliflower leaves impregnated with 0.002% of imidacloprid
resulted in decrease of net reproductive rate (RQ) of a surviving adult
from 10.99 to 8.47 females/female. The potential fecundity (Pf) was
-10-
^ItfitnewofLftemhtre
significantly decreased to 166.5, wliereas, intrinsic rate of increase (r^)
was 0.073 females/female/day as compared to 0.09 in untreated
control.
Rathod et ai, (2003) conducted an experiment in India during
1997-2000 to determine the efficacy of imidacloprid against jassids
{Amrasca biguttula), aphids {Aphis gossypii), and thrips {Thrips tabaci)
infesting cotton. The treatments comprised imidacloprid at 5.0, 7.5, and
10 g/kg; diafenthiuron 300 and 400 g/ha; and dimethoate at 1.25
litres/ha. The lowest mean population of jassids (0.99), aphids (4.41),
and thrips (1.73) per 3 leaves were obtained with 10 g imidacloprid/kg,
300 g diafenthiuron/ha. and 5 g imidacloprid/kg, respectively. The
highest cotton yield (826 kg/ha) was obtained with 5 g imidacloprid/kg.
Hu and Prokopy (1998) tested imidacloprid in the laboratory as
well as in the field against Rhagoleta pomonella flies to determine
ingestion/ contact or contact alone toxicity and reported that
imidacloprid 10-12 times more toxic and acted more rapidly by oral
ingestion than by surface contact. Affected flies were observed to cease
feeding and then regurgitate. Mortality stabilized 4 days after treatment.
Compared with control flies, females exposed to imidacloprid showed
reduced fecundity regardless of whether exposure was by oral or
surface contact. In field experiments, spray applications of imidacloprid
to foliage at the manufacturer's recommended rate resulted in no
-11-
^fview of Literature
significant mortality of flies, either among flies released immediately
after treatment or 24 h later.
Imidacloprid residues upto 2ppm in soil did not affect egg viability
or days to hatch of Japanese beetle Popillia Japonica, but they killed
neonates soon after eclosion (George et al., 2007).
Bao et al., 2009 tested four insecticides against brown plant
hopper and reported that imidacloprid and dinotefuran reduced the
fecundity of BPH to 68.8% and 52.4% in macropterous families, and
57.9% and 43.1% in brachypterous families, as compared with the
untreated controls, by contrast, triazophos and fenvalerate increased
the fecundity.
Christopper et al., (2009) observed that when Green peach aphid,
Myzus persicae (Sulzer) were exposed to potato leaf discs dipped in
sublethal concentrations of insecticide, almost all measured endpoints-
adult longevity, F1 production, F1 survival and F2 production-were
affected, and a statistically significant (P < 0.05) stimulatory response
was recorded for F2 production following exposure to imidacloprid.
Boina et al., (2009) studied the effects of sublethal concentrations
of imidacloprid on life stages of Asian citrus psyllid (ACP), Diaphorina
citri and reported that feeding by ACP adults and nymphs on plants
treated daily with a sublethal concentration (0.1 microg mL" ) of
imidacloprid significantly decreased adult longevity (8 days), fecundity
-12-
^fviewofLiUrature
(33%) and fertility (6%), as well as nymph survival (12%) and
developmental rate compared with untreated controls. The magnitude of
these negative effects was directly related to exposure duration and
concentration. Furthermore, ACP adults that fed on citrus leaves treated
systemically with lethal and sublethal concentrations of imidacloprid
excreted significantly less honeydew (7-94%) compared with controls in
a concentration-dependent manner suggesting antifeedant activity of
imidacloprid. It has been concluded from the experiment that sublethal
concentrations of imidacloprid negatively affect development,
reproduction, survival and longevity of ACP, which likely contributes to
population reductions over time. Also, reduced feeding by ACP adults
on plants treated with sublethal concentrations of imidacloprid may
potentially decrease the capacity of ACP to successfully acquire and
transmit the HLB causal pathogen.
Banerjee et al., (2001) studied the growth regulatory effect of
naturally occurring quinines and their derivatives against D. koenigii
and reported that all the test compounds disrupted the normal growth at
sublethal doses while 2,6-dimethylbenzoquinone and menadione at low
doses. Baised on LD50 values, the following increasing order of toxicity
could be established: plumbagin < 2, 6-dimethylhydroquinone < 2,3,6-
trimethylbenzoquinone<juglone<menadione<2,6dimethylbenzoquinone.
-13-
^^gviewofLiUrahtn
Amarasekare and Edelson (2004) evaluated the efficacy of
diflubenzuron, azadirachtin, Beauveria bassiana, spinosad, endosulfan,
esfenvalerate, and naled to third instars of differential grasshopper,
Melanoplus differentialis (Thomas), at temperatures ranging from 10 to
35°C. In the laboratory, treatment with esfenvalerate resulted in 100%
mortality at temperatures of 10 to 35X, and efficacy was not
temperature dependent. Treatment with spinosad resulted in similar
mortality as with esfenvalerate at all temperatures except lOX. The
activity of B. bassiana was greatest at 25°C and was adversely affected
by high and low temperatures.
Gregory et al., (2005) studied the persistence and efficacy of
spinosad against Rhyzopertha dominica (F.) in wheat stored for 9
months at 30°C temperature and 55 and 70% relative humidity, and
reported that spinosad applied at 0.5 or 1 mg kg" was completely
effective for 9 months, with 100% adult mortality after 14 days of
exposure and no live Fi adults produced. Adult mortality was <100% in
some samples of wheat treated with 0.1 mg kg" of spinosad, and live
progeny were produced in all samples treated at this level. Further, they
concluded that spinosad is likely to be an effective grain protectant
against R. dominica in wheat stored in warm climates.
Behnam Amiri-Besheli (2009) determine the efficacy of Tracer
(Spinosyn), Insecticidal Gel (Palizin), Insecticidal Emulsion (Sirinol),
-14-
^^fviewofLittmiun
Runner (Methoxifenozide) and Tondoxir with and without mineral oils
(MO) for the control of citrus leaf-miner (CLM), Phylocnistis citrella
Stainton (Lepidoptera: Gracillariidae) in laboratory condition at 24, 48,
72 and 96 h post treatments. Leaves of citrus with second and third
instars of leaf-miner larvae were used in all tests and the larval mortality
were monitored. Analysis of variance showed that there were significant
differences between treatments and control, and also significant
differences were found among treatments. Tukay-Test among above
treatments has shown that Tracer + oil, Runner and Runner + oil and
Tondxir + oil with 98 ± 3. 2, 98 ± 8.1 and 93 ± 5% mortality were more
effective than MO, Tondxir and Sirinol + oil with 85 ± 8.3 and 81 ±7.2 and
78.25 ±8.2% mortality and significant different with control (P< %1),
respectively. Tukay-Test among 96 and 72h post treatments with 77 ±
3.82 and 70.65 ± 7.5% of total mortality are more effective than 48 and
24 h post treatments with 52.27 ± 4 and 35.90 ± 3.8% of total mortality
respectively
Bill et al., (2009) assessed the combining effects of spinosad at
0.01, 0.1 and 0.5 ppm, with the diatomaceous earth (DE) formulation
SilicoSec at rates of 150, 300 and 600 ppm against larvae and adults of
three different populations of Tribolium confusum du Val (Coleoptera:
Tenebrionidae), originating from different European countries (Greece,
Portugal and Denmark). Tests were conducted on wheat and maize at
•15-
^tfviewafLiUmtttre
25 and 30°C and survival of larvae was assessed after 7 d exposure
and survival of adults was assessed after 7 d and 14 d of exposure. At
each dose of spinosad, survival of T. confusum individuals decreased
as the rate of DE increased. As temperature increased, the efficacy of
spinosad and Silicosec alone or in combination also increased. The
efficacy of spinosad alone was slightly higher on maize than wheat,
while the reverse was noted for all the tested combinations of spinosad
with DE as well as in the case of the application of DE alone. The strain
from Portugal was always the least susceptible of the three tested.
Xian-Hui et ai, (2008) investigated the effects of sublethal
concentrations of spinosad applied for 48 h at LC25 (0.12 mg L"^) and
LC50 (0.28 mgL"^) on the biological characteristics i.e. development
time, pupation rate, pupal weight, fecundity, fertility, larval weight and
survival rate in the parent and offspring, of the diamondback moth,
Plutella xylostella (Lepidoptera: Yponomeutidae) and reported that the
pupation rate and pupal weight were significantly lower in treated
groups in which the third instars were treated with spinosad at LC25 or
LC50 than in the control group. The fecundity, egg size and reproductive
effort (fecundityxegg size) were strongly reduced, and the hatchability of
smaller eggs tended to be lower in treated groups. The survival rate of
the offspring at immature stages was lower in the LC25 and LC50
treatment groups than in the control group, and the development time
-16-
^^fviewofLiUmture
tended to be prolonged in the treatment groups. The mean values of the
intrinsic rate of increase (rm), finite rate of increase (D), gross
reproductive rate (GRR) and net reproductive rate (Ro) were
significantly lower in the treatment groups than in the untreated group.
Generally, the effects on biological characteristics were greater in the
LC50 treatment group, and the effects on the offspring were inuch
smaller than those on the parent generation.
Khowaja et al., (1994) studied the effects of sub-lethal
concentrations (10, 15, 20, 25 and 30 ppm) of monocrotophos applied
topically on 5th-instar nymphs of Dysdercus cingulatus Fabr. in the
laboratory at controlled conditions (29 ± VC and 70-80% R.H.) and
reported that the longevity and weight of the treated nymphs increased
linearly and showed positive correlation. On the other hand, fecundity
and fertility of the adults emerging from the treated nymphs decreased
linearly and showed negative correlation with the increasing sub-lethal
concentrations. The LD 50 value representing a very small percentage
of monocrotophos (30 ppm/nymph) increased 5th-instar nymphal
longevity by 43.14%, whereas, the longevity of the adult males and
females enhanced by 31.48 and 26.50%, respectively as compared to
control. Similarly, the weight of the 5* instar treated nymphs increased
by 15.9%; but adult males and females gained 7.9 and 11.8% more
weight as compared to control. On the contrary, fecundity and fertility of
-17-
^ftfiewofLitemtun
the females emerging from the surviving nymphs declined by 18.1 and
22.4%, respectively as compared to controls.
Khowaja et al., (1996) observed that the fecundity and fertility of
the emerging females of Dysdercus cingulatus following the topical
application of 6, 4, 2, 1, and 0.5 Dg doses of two analogues of D-
ecdysone (22, 25-dideoxyecdysone and 2, 22, 25-tndeoxy-5
D-hydroxy ecdysone) decreased linearly and showed negative
correlation with the increasing dose levels. After the application of the
highest dose (6 Dg/nymph) of 22,25-dideoxyecdysone, the maximum
reduction in the egg production and egg hatching of the females that
emerged from the treated nymphs was 30.85 and 35.28%, respectively
as compared to control. But, the same dose of 2,22,25-trideoxy-5n-
hydroxyecdysone resulted in 27.55 and 29.18% reduction in fecundity
and fertility of the females which was 3.3 and 6.1% less when compared
with 22,25-dideoxyecdysone, respectively.
Khowaja and Qamar (2003) observed by topical application of
sublethal concentrations (10, 15, 20, 25 and 30 ppm) of monocrotpphos
on the individual 4th instar nymphs of D. cingulatus resulted in
concentration-based changes. The regression between concentration
strength and average longevity showed a positive correlation. The life
span of the treated 4th instar nymphs as well as those of the next instar
and adult males and females increased linearly, whereas, the weight of
-18-
^StfviewofCiUmtun
the treated nymphs as well as those of ensuing stages did not change
significantly compared to the control. However, the egg production of
the females emerged from the treated nymphs and egg hatching
decreased linearly and showed a negative correlation with increasing
concentrations. Furthermore, the anatomical structure of gonads of the
insects was also affected. The application of higher concentrations, i.e.
25 and 30 ppm, resulted in the gradual degeneration of oocytes, which
ended up in total dissolution of the ovarian follicles. Eventually, all the
oocytes degenerated completely. The lower concentrations of
monocrotophos did not change the morphological and cellular structure
of the gonads, which remain more or less unchanged.
Khowaja et al., (2001) examined the sublethal levels of cythion
(malathion) at the concentrations of 10, 20, 40, 60 and 80 ppm/4th
instar nymph of Dysdercus koenigii resulted in concentration-based
changes. The nymphal loss and survival duration of the treated insects
were directly proportional and increased linearly with the increase in
concentration. On the other hand, the fecundity and fertility of the
adults, emerged from the treated nymphs were inversely proportional,
decreased linearly and showed negative correlation with concentrations.
The initial mortality following the topical application of higher
concentrations (i.e. 80, 60 and 40 ppm) was very high within 24 hours.
However,, the subsequent mortality up to adult emergence was
-19-
^^fmwofLitemture
insignificant. Out of the total adults that emerged from the treated
nymphs, few were with malformed wings and legs. The lower
concentrations (i.e. 20 and 10 ppm) had negligible effect and were well
tolerated.
-20-
MATERIAL AND METHODS
Breeding and maintenance of stock culture of Dysdercus koenigii
Adults of Dysdercus koenigii were collected from the cotton field
during the kharief period and were kept in glass jars measuring
20x15cm, containing 2-3cm thick layer of moist and loose sterilized
sand at the bottom. These jar were kept at two different temperatures
with photoperiod regime, viz. 25±1°C / 12L: 12D and 30 ± 1°C / 14L:
10D in different B.O.D incubators where as the relative humidity was
kept constant in both incubators i.e. 60-70%R.H. These insects were
fed on water soaked healthy cotton seeds, which were changed daily.
After egg laying, the adult insects were transferred to fresh rearing jars
to avoid overcrowding. On hatching, the nymphs were daily provided
with soaked healthy cotton seeds.
Sampling of experimental insects
Newly moulted one day old 4** instar nymphs of Dysdercus
koenigii were used for the experiments. The samples of freshly moulted
3 ^ instar nymphs between 11:00am and 12:00 noon each day were
isolated and maintained age wise. The experiments were run under
controlled laboratory conditions at two different temperature and
photoperiod i.e. 25±1°C / 12L: 12D and 30 ± 1°C / 14L: 10D where the
relative humidity was kept constant in both cases. Subsequently, fresh
MaUmCand9ietfu)ds
moulted 4* instar nymphs of Dysdercus koenigii were treated with
different concentrations of selected insecticides for observation as
described below.
Insecticides Used
1. Confidor (Imidacloprid 17.80% SL)
2. Tracer (Spinosad 45%SC)
These chemicals were purchased commercially from the authorized
dealer.
Dilution and application of insecticides
The following formula was proved helpful to make the stock
solution as
X = AxB/C
Whereas
X = Volume of the solution in hand to be diluted to desired volume
A = Concentration desired
B = Volume of solution desired
C = Concentration of solution in hand.
The stock solution of each insecticides were further diluted to sub
lethal concentration of each insecticide on the basis of LD 30 value as,
Imidacloprid: 0.008%, 0.006%, 0.004%, 0.002%, 0. 001%
Spinosad: 0.1%, 0.08%, 0.06%, 0.04%, 0.02%.
-22-
Material and Methods
These sub lethal concentrations of each insecticide were used for
topical application on the 4* instar stage of Dysdercus koenigii. The
effect of these chemicals was observed on mortality, malformation,
fecundity, fertility and longevity by using five-sub lethal concentrations
of each insecticide at their respective temperature and relative humidity.
For each concentration of diluted insecticide only 1|j| was applied
topically on the thoracic pleuron of individual IV** instar of Dysdercus
koenigii by 26-gauge needle attached to a tuberculin syringe fitted into a
manually operated micro applicator. At the time of application of
insecticides the age of the experimental insect was one day. For each
concentration and temperature with their photoperiod 30 individual IV'
instar nymphs were used. Two controls at each temperature,
photoperiod and humidity consisting of same number of nymphs of
same age were also run parallel to each series of experiment. One
control was treated with acetone (solvent) and served as control-l while
untreated nymphs of the same age served as control-l I. The controlled
groups are maintained at their respective controlled conditions. When
females subsequently emerged from the treated nymphs they were
paired with the males emerged from the untreated nymphs of
corresponding age from control II (Magdum et al., 2001). The data on
mortality, malformation, fecundity, fertility and longevity were recorded
on both treated and control insects.
-23-
Material aiuf94etfukb
Method of observation
Following the application of each concentration of insecticide on
24-hour-olcl 4'^ instar nymphs of D. koenigii. Observations were made
on mortality, malformation, longevity as well as fecundity and fertility of
females emerged from the treated nymphs. The data was compared
with acetone treated control (control-1). The selection of concentrations
was made on the basis of LD30 value. For determining LD30 value of
insecticides different concentration was prepared and applied topically
by micro applicator. For observations on the fecundity and fertility pf the
females, the adult males emerged from the survived treated nymphs
were paired with the affected females of the corresponding age and
stage and each pair was kept in separate rearing jar at controlled
conditions. The number of eggs laid by each female and fertility were
recorded. The unhatched eggs were also counted. Similar observations
were also made in controls consisting of the males and females
emerged from acetone-treated as well as untreated nymphs.
Interpretation of data
The data on mortality, malformation, fecundity, fertility and
longevity of Dysdercus koenigii affected by different concentrations of
various insecticides were recorded for each replicate and mean value of
four such replicates was calculated. The determination of any change
was not only ascertained arithmetically but appreciated by statistically
-24-
MaUfiaC and Methods
analysis also. The standard deviation (S.D) was calculated by the
following formula
S.D.= V^Z)'/«-l
Where,
S.D = Standard deviation
D = Sum of squares of the difference of the mean value.
n = Number of observation
On the basis of S.D (Standard deviation), Standard error was
calculated by the formula as
5£.= S.DJ 4n
Where,
SB = Standard error
S.D = Standard deviation
n • = Number of observation
It was necessary to appreciate the difference between mean
values of different sets of samples. Arithmetically it was uncertain;
therefore we tested the mean value statistically for this purpose the
statistical technique of variation significance (t-Test) was applied
(Bailey-1959) by using the formula
SD, m L 4- 1 «,
-25-
MaUmCan^Metfukb
Where,
t = Significant value
mi = Mean value of first set of observation
m2 = Mean value of second set of obsen/ation
SDi = Standard deviation of first set of observation
SD2 = Standard deviation of second set of observation
ni = Number of observations of the first set
n2 = Number of observations of the second set
The calculated 't' was compared with tabulated 't' (Bailey- 1959)
at 5% level. If the calculated value remained higher than the tabulated 1'
value, the data were regarded significant otherwise insignificant. The
tabulated value oft' at 5% level is 2.447.
Further, to measure the intensity of association between mortality,
malformation, fecundity, fertility and concentration regression line was
fitted to the data, followed by co-relation analysis (Sokal and Rohlf,
1981) using the formula
Y= A+Bx
-26-
RESULTS AND OBSERVATION
Efficacy of Imidacloprid at 30±rc temperature, 14L: 10D
photoperiod and 60-70%RH
For control-1 treatment, 4*' instar nymphs of same age were
sorted out from the culture and treated topically with 1pl acetone while,
for control-!! nymphs of the corresponding age remained untreated. No
significant effects were observed on comparing acetone treated nymphs
{control-1) with untreated nymphs (control-ll). Mortality was not seen
upto the adult emergence during the course of control experiments
(Table 1). The female emerged from acetone treated nymphs, when
mated with normal males, laid almost equal number of eggs to those
emerged from the untreated nymphs. Similarly, there was no significant
effect on the fecundity (t = 1.180, P > 0.05) and fertility (t = 2.130, P >
0.05). The longevity recorded during 4* and 5* nymphal instars as well
as adult male and adult female life stages was 5.25± 0.24, 6.75± 0.24,
20.25 ± 0.24 and 16.75± 0.24 days respectively.
Different sublethal concentration of imidacloprid i.e. 0.001%,
0.002%, 0.004%, 0.006%, and 0.008% were applied topically on 24-
hour-old 4** instar nymphs of Dysdercus koenigii. The number of insects
taken for each treatment was thirty and the experiment was repeated
three times. Each insect was treated topically with 1 | j | of these sublethal
concentrations of insecticide by 26-guage needle attached to a
^l(psu&s and^OSs&vation
tuberculin syringe fitted into a manually operated micro applicator. The
insecticide was applied on the thoracic pleuron of individual 4* instar
nymphs. Observations were made on various aspect of biology like
mortality, malformation, fecundity, fertility and longevity.
Effect on Mortality
The mortality was recorded at two stages, initial mortality upto 24
hour and the total mortality upto adult emergence. The initial nymphal
mortality upto 24 hours at five different sublethal concentration viz.,
0.001%, 0.002%, 0.004%, 0.006% and 0.008% were recorded as 3.33 ±
0.33 (t = 4.186, P > 0.05), 4.00 ± 0.57(t =3.464, P > 0.05), 6.66 ± 0.33(t
= 5.920, P > 0.05), 8.66 ± 0.33(t =6.751, P > 0.05) and 9.00 ± 0.57 (t =
5.196, P> 0.05) respectively. The results were found to be statistically
significant when compared with the acetone treated (Control-I). The
total nymphal mortality upto adult emergence following the topical
application of above said sublethal concentration were recorded as 3.66
± 0.33 {t = 4.388, P > 0.05), 6.00 ± 0.57{t = 4.242, P > 0.05), 10.66 ±
0.33 (t = 7.490, P > 0.05), 12.66 ± 0.33 (t = 8.162, P > 0.05) and 14.00 ±
0.57 (t = 6.480, P > 0.05) respectively, which were found to be
statistically significant as compared with control-1. The initial mortality
upto 24 hours and total mortality upto adult emergence of the treated
nymphs increased linearly with increasing sublethal concentration of
imidacloprid at 3 0 ± r c temperature, 14L:10D photoperiod and 60-
-28-
^^§su&s aiufOSservation
70%RH (y = 1004.4X + 1.910, R = 0.878, P > 0.001 and y = 1624.7x +
2.387, R- = 0.918, P > 0.001 respectively) which shows positive
correlation coefficient (Graph land 2).
Effect on Malformation
Malformations in the nymphs and adults emerged from treated 4*
instar nymphs of D. koenigii were of variable degree in the whole body,
wings and legs. A number of adults were found having folded, twisted
and deformed wings as well as sunken abdomen. In controlled sets, the
malformations were not observed. The malformation either in wings or
legs caused by these sublethal concentrations viz., 0.001%, 0.002%,
0.004%, 0.006% and 0.008% were recorded as 2.66 ± 0.33 (t = 3,741,
P > 0.05), 3.33 ± 0.33 (t = 4.186 P > 0.05) 3.33 ± 0.33 (t = 4.186 P >
0.05), 3.66 ± 0.33 (t = 4.186, P > 0.05) and 5.00 ± 00 (t = 6.388, P >
0.05) respectively (Plate-1). These results were statistically significant
when compared with the acetone treated nymphs or untreated nymphs.
The malformation caused due to treatment of nymphs increased linearly
with increasing sublethal concentration of imidacloprid (y = 417.14x +
1.599, R = 0.665) showing positive correlation coefficient (Graph 3).
The malformations were much pronounced in higher concentrations.
The abnormal adults formed were found to have shorter life span.
-29-
^Stpsu&s andOSservation
Effect on Fecundity and Fertility
The Fecundity and Fertility were affected by sublethal
concentration of insecticides viz., 0.001%, 0.002%, 0.004%, 0.006%,
and 0.008% and fall in fecundity were recorded as 179.75 ± 1.49 (t =
4.418, P > 0.05), 144.25 ± 1.49 (t = 5.853, P > 0.05), 110.75 ± 0.85 (t =
7.454, P > 0.05), 92.00 ± 1.29 (t = 8.092, P > 0.05) and 83.25 ± 0.85 (t =
8.291, P > 0.05) at respective concentrations as compared to control
whereas, the fertility were recorded as 154.50 ± 1.04 (t = 6.902, P >
0.05), 122.25 ± 1.24 (t = 5.853, P > 0.05), 86.75 ± 0.85 (t = 10.725, P >
0.05), 66.00 ± 0.91 (t = 11.441, P > 0.05) and 52.25 ± 1.24 (t = 11.200,
P > 0.05) respectively. Both the results were statistically significant
when compared with control-1. The results showed reduction in
fecundity and fertility on increasing the concentration of imidacloprid. At
lowest selected concentrations the fecundity dropped to 20.72% while
the fertility / egg hatching were reduced to 26.42%. Similarly, at highest
selected concentration the fecundity dropped to 63.28% and fertility
reduced to 75.11%. The average egg production of the females that
emerged from treated 4*'' instar nymphs decreased linearly with
increasing sublethal concentration (y = -15864x + 192.6, R = 0.855)
showing negative correlation coefficient (Graph 4) while the fertility of
the eggs laid by the affected females also decreased linearly (y = -
-30-
^^fsu&s awCOBsefwition
17018X + 172.3, R = 0.858) and showed negative correlation
coefficient (Graph 5).
Effect on Longevity
The effect of topical application of imidacloprid on longevity has
been recorded during nymphal instar (4* and S*" ) and adult (male and
female) stage of Dysdercus koenigii. The longevity for 4 ^ nymphal instar
at sublethal concentrations viz., 0.001%, 0.002%, 0.004%, 0.006% and
0.008% were 5.50 ± 0.28 (t = 0.975, P > 0.05), 5.75 ± 0.24 (t = 1.428, P
> 0.05), 6.00 ± 00 (t = 2.474, P > 0.05), 6.50 ± 0.28 (t = 5.182, P > 0.05)
and 6.75 ± 0.24 (t = 2.474, P > 0.05) days respectively. The result at
lower concentration (0.001% and 0.002%) showed statistically
insignificant increase in nymphal duration whereas, at higher
concentration (0.004%, 0.006% and 0.008%) gave significant when
compared with the control-1. Similarly, the 5* nymphal instar span were
7.25 ± 0.24 (t = 1.428, P > 0.05), 7.75 ± 0.24 (t = 2.020, P > 0.05), 8.00
± 00 (t = 3.194, P > 0.05), 8.50 ± 0.28 (t = 2.581, P > 0.05) and 8.75 ±
0.24 (t = 2.857, P > 0.05) days at above mentioned concentrations
respectively. The results at lower concentrations i.e. 0.001% and
0.002% gave insignificant increase whereas higher concentration i.e.
0.004%, 0.006%, and 0.008% gave statistically significant enhancement
when compared with control-1. The adult male longevity were recorded
at these concentrations as 21.75 ± 0.24 (t = 2.474, P > 0.05), 22.00 ± 00
-31-
^^fisufts andOSsetvation
(t = 3.779, P > 0.05), 22.75 ± 0.24 (t = 3.194, P > 0.05), 23.50 ± 0.28 (t
= 3.761, P > 0.05) and 24.00 ± 0.28 (t = 5.532, P > 0.05) days
respectively. The results were found to be significant when compared
with control-1. The adult female longevity at these concentrations were
recorded as 17.00 ± 00(t = 1.428 P > 0.05), 17.25 ± 0.47 (t = 1.428, P >
0.05), 18.50 ± 0.28 (t = 2.569, P > 0.05), 20.00 ± 00 (t = 5.150, P >
0.05) and 20.50 ± 0.28 (t = 3.761, P > 0.05) days respectively. The
results were found to be significant at higher concentration (0.008%,
0.006%, and 0.004%) but were insignificant at lower concentrations
(0.001% and 0.002%). The longevity of treated 4"" instar nymphs
increased linearly with increasing sublethal concentrations and same
affects were observed in rest of the life stages as 4'* instar (y = 176.3x +
5.3677, R2 =0.976), 5* instar (y = 223.5x + 7.084, R = 0.929), adult
male (y = 387.9x + 21.07, R =0.872) and adult female (y = 487.9x +
16.69, R? = 0.9676) shows positive correlation with increasing
concentrations (Graph 6-9).
Efficacy of Imidacloprid at 25±1*C temperature, 12L: 12D
photoperiod and 60-70%RH
For control set I, when the 4* instar nymph was treated topically
with 1 1 acetone solution no mortality was observed upto adult
emergence (Table 2). For control set II the initial mortality upto 24
hours, total mortality upto adult emergence, malformation, fecundity,
-32-
^§su&s andOBsewation
fertility and longevity were compared with the acetone treated control I.
The ecdysed 5' ^ instar nymphs and as well as emerged adult male and
female were normal in appearance showing no malformation. The adult
females emerged from the acetone treated 4th instar nymphs niated
with the untreated normal males (control- II), laid almost equal number
of eggs as compared to females emerged from the untreated 4 * instar
nymphs (Table 3), showing no significant difference in fecundity (t =
1.052, P > 0.05) and fertility (t = 1.973, P >0.05). The longevity recorded
during 4* and 5* nymphal instar as well as adult males and adult
female stage were 6.50 ± 0.28, 6.50 ± 0.28, 22.25 ± 0.24 and 17.50 ±
0.28 days respectively. The survival duration of all the stages in both
control I and control II remain almost same indicating no statistical
difference.
Effect on Mortality
The initial mortality upto 24 hours and total mortality upto adult
emergence were recorded with treated 4* instar nymphs of D. koenigii.
The 4'^ instar nymphs were treated with different sublethal
concentrations as, 0.001%, 0.002%, 0.004%, 0.006% and 0.008% of
imidacloprid to observe the initial nymphal mortality upto 24 hours and
total mortality upto adult emergence. The initial mortality upto 24 hours
were recorded at these sublethal concentrations as, 2.33 ± 0.33(t =
3.501, P > 0.05), 3.00 ± 0.57 (t = 3.000, P > 0.05), 5.00 ± 0.57 (t =
-33-
^S^fsufts cmdOSsmmtion
3.872, P > 0.05), 5.33 ± 0.33 (t = 5.292, P > 0.05) and 7.00 ± 0.57 (t =
4.542, P > 0.05) respectively. The results were found to be significant
when compared with control-1. The total nymphal mortality upto adult
emergence were found to be 3.33 ± 0.33 (t = 4.186, P > 0.05), 6.00 ±
0.57 (t = 4.242, P > 0.05), 9.00 ± 0.57 (t = 5.196, P > 0.05), 11.00 ± 0.57
(t = 5.744, P > 0.05) and 12.00 ± 0.57 (t = 6.000, P > 0.05) respectively.
The results came to be statistically significant when compared with
control-1. The initial mortality of nymphs upto 24 hours and total
mortality upto adult emergence at this temperature and relative humidity
increased linearly with increasing sublethal concentrations (y = 727.1x +
1.340, R = 0.891 and y = 1364.5x + 2.3171, R = 0.904, P > 0.01
respectively) showing positive correlation coefficient (Graph 10 and 11).
Effect on Malformation
The affects on malformations of variable degree were noticed in
the wings, legs and body of adults that emerged from the treated 4*
instar nymphs. The malformation formed at sublethal concentration viz.,
0.001%, 0.002%, 0.004%, 0.006% and 0.008% were found to be 0.67 ±
0.33 {t = 1.877, P > 0.05), 2.33 ± 0.33 (t = 3.501, P > 0.05), 3.00 ± 00 (t
= «°, P > 0.05), 3.33 ± 0.33 (t = 4.186, P > 0.05), and 4.33 ± 0.33 (t =
4.773, P > 0.05) respectively {Plate-2). There was no significant effect
on malformation at 0.001% and 0.004% concentration while treatment
with 0.008%, 0.006% and 0.002% indicate statistically significant results
-34-
^^^ and observation
as compared to control-1. The proportion of malformation increased on
increasing tiie concentration of insecticide (y = 485.7x + 0.649, R =
0.912). Thus malformation showed positive correlation with increasing
sublethal concentration at 25±1°C/ 12L: 12D temperature, photoperiod
and constant relative humidity {Graph 12).
Effect on Fecundity and Fertility
The fecundity were recorded at sublethal concentrations viz.,
0.001%, 0.002%, 0.004%, 0.006% and 0.008% as 157.75 ± 1.54 (t =
3.762, P > 0.05), 128.50 ± 1.70{t = 4.687, P > 0.05), 103.75 ± 1.88 (t =
5.300, P > 0.05), 70.75 ± 0.62 {t = 6.692, P > 0.05) and 65.25 ± 0.62 {t =
6.824, P > 0.05) eggs respectively. While, the egg hatching at these
sublethal concentration were recorded as 132.50 ± 1.70 {t = 6.049, P >
0.05), 105. 50 ± 0.28 (t = 9.396, P > 0.05), 76.50 ± 1.49 (t = 8.954, P >
0.05), 46.25 ± 0.62 (t = 12.492, P > 0.05) and 40.00 ± 0.91 (t = 11.854,
P > 0.05) respectively. The results were statistically significant when
compared with the control-l. The fecundity decreased linearly with
increasing sublethal concentrations (y = -15623x + 174.5, R = 0.868)
showing negative correlation coefficient (Graph 13), similarly the fertility
also reduced linearly (y = -15827x + 150.6, R = 0.863) with increasing
sublethal concentrations and shows negative correlation coefficient
(Graph 14).
-35-
^fstdts andOBsermtion
Effect on Longevity
The longevity was reported at three life stages such as, 4*^ S**"
nymphal instars as well as adult male and female. The longevity of 4*
instar nymphs at sublethal concentrations viz., 0.001%, 0.002%,
0.004%, 0.006% and 0.008% were recorded as 6.75 ± 0.24 (t = 0.971,
P > 0.05), 7.00 ± 00 (t = 1.873, P > 0.05), 7.50 ± 0.28 (t = 1.873, P >
0.05), 8.00 ± 00 (t = 3.244, P > 0.05) and 8.25 ± 0.24 (t = 2.564, P >
0.05) days respectively. The results at lower concentrations i.e. 0.001%,
0.002% and 0.004% were statistically insignificant while at higher
concentrations i.e. 0.006% and 0.008% these were statistically
significant. The longevity were recorded for 5* nymphal instar at these
concentrations were 7.00 ± 00 (t = 1.873, P > 0.05), 7.75 ± 0.28 (t =
1.428, P > 0.05), 8.50 ± 0.28 (t = 2.649, P > 0.05), 8.75 ± 0.24 (t =
2.913, P > 0.05) and 9.50 ± 0.28 (t = 3.244, P > 0.05) days respectively.
These results were statistically insignificant at 0.001% and 0.002%
while higher concentrations i.e. 0.004%, 0.006% and 0.008% gave
statistically significant results. The adult male longevity were recorded
as 23.50 ± 0.28 (t = 2.171, P > 0.05), 24.25 ± 0.24 (t = 2.857, P > 0.05),
25.00 ± 00 (t = 4.738, P > 0.05), 25.75 ± 0.24 (t = 3.779, P > 0.05) and
26.50 ± 0.28 (t = 4.004, P >0.05) days respectively. The results were
statistically insignificant at 0.001% while at 0.002%, 0.004%, 0.006%
and 0.008% showed statistically significant. The adult female longevity
-36-
^fsulis amfOSsewation
was recorded as 17.75 ± 0.24 (t = 0.971, P > 0.05), 18.00 ± 00 (t =
1.873, P > 0.05), 18.50 ± 0.28 (t = 1.873, P > 0.05), 19.00 ± 00 (t =
3.244, P > 0.05) and 19.50 ± 0.28 (t = 2.649, P > 0.05) days
respectively. These gave insignificant result at 0.001%, 0.002% and
0.004% whereas, at higher concentrations i.e. 0.006% and 0.008% it
was significant. The longevity of treated 4 ^ instar (y = 214.Ox + 6.616,
R > 0.981), 5* instar (y = 339.8x + 6.861, R = 0.954), adult male (y =
456.8X + 23.011, R =0.921) and adult female (y = 236.1x +17.58, R =
0.989) increased linearly with increasing sublethal concentrations of
imidacloprid showing positive correlation coefficient (Graph 15-18)
Efficacy of Spinosad at 30±1*'C temperature, 14L: 10D photoperiod
and 60-70%RH
Different aspects of biology as mortality upto 24 hours, mortality
upto adult emergence, malformation, fecundity and fertility and longevity
were recorded. No significant effects were found while comparing the
acetone treated control-1with untreated control-ll. The initial mortality
upto 24 hours and total mortality upto adult emergence and
malformation were not found during different stages of life cycle. The
fecundity and fertility were recorded as 226.75 ± 3.32 (t = 1.180, P >
0.05) and 210.00 ± 1.29 (t = 2.130, P > 0.05) respectively. The
untreated females laid 231.25 ± 3.06 numbers of eggs and the fertility
was recorded as 218.25 ± 2.35. The longevity of treated 4**" instar and
-37-
^ifisu&s otuCOSsmHition
5' instar nymphs, adult male and adult female were recorded as 5.25 ±
0.24, 6.75 ± 0.24, 20.25 ± 0.24 and 16.75 ± 0.24 days respectively and
no effect was found while comparing acetone treated nymphs (control-l)
with untreated ones (control-ll).
Effect on Mortality
The initial mortality of nymphs upto 24 hours at sublethal
concentrations viz., 0.02%, 0.04%, 0.06%, 0.08% and 0.1% were
recorded as 0.66 ± 0.33 (t = 3.473, P > 0.05), 2.33 ± 0.57 (t = 3.501, P >
0.05), 5.00 ± 0.57 (t = 3.872, P > 0.05), 7.33 ± 0.33 (t = 6.211, P >
0.05), 8.33 ± 0.33 (t = 6.621) respectively. The results were statistically
significant when compared with control-1. The total mortality upto.adult
emergence was also recorded as 1.66 ± 0.33 (t = 2.955, P > 0.05), 7.00
± 0.57 (t = 4.582, P > 0.05), 10.66 ± 0.33 (t = 7.490, P > 0.05), 13.33 ±
0.33 (t = 8.376, P > 0.05), and 17.00 ± 0.57 (t = 7.141, P > 0.05)
nymphs respectively. The results were statistically significant when
compared with control-1. The initial mortality upto 24 hrs and total
mortality upto emergence of treated 4*'' instar nymphs increased linearly
with increasing sublethal concentration (y = 91.9x - 0.653, R = 0.973
and y = •176.6x - 0.558, R = 0.987 respectively) showing positive
correlation coefficient (Graph 19 and 20).
-38-
^^fisu&s ondOSserMtim
Effect on Malformation
The malformation were recorded at sub lethal concentrations of
spinosad viz., 0.02%, 0.04%, 0.06%, 0.08% and 0.1% as 6.00 ± 0.57 (t
= 4.242, P > 0.05), 5.00 ± 00 (t = 3.501, P > 0.05), 5.66 ± 0.33 (t =
5.457, P > 0.05), 4.33 ± 0.33 (t = 4.773, P > 0.05), 5.33 ± 0.33 (t =
5.296) respectively (Plate-3). The results came to be statistically
significant when compared with control-1. The malformation during their
life stages increased linearly with increasing sublethal concentration (y
= 31.85X .+ 2.793, R = 0.287) showing positive correlation coefficient
(Graph 21)
Effect on Fecundity and Fertility
The number of eggs (aid by the affected females at sublethal
concentration viz., 0.02%, 0.04%, 0.06%, 0.04% and 0.1% were 161.50
± 1.32 (t = 5.300, P > 0.05), 138.75 ± 1.24 (t = 6.202, P > 0.05), 130.00
± 1.95 (t = 6.053, P > 0.05), 118.75 ± 2.24 (t = 6.224, P > 0.05) and
103.75 ± 1.88 (t = 6.878, P > 0.05) respectively. The egg hatching was
also recorded on these concentrations as 142.50 ± 0.64(t = 7.942, P >
0.05), 117.25 ± 0.85(t = 9.302, P > 0.05), 106.25 ± 0.85(t = 9.830, P >
0.05), 90.75 ± 0.33 (t = 9.979, P > 0.05), and 72.50 ± 1.04 (t = 10.863, P
> 0.05) respectively. These results came to be statistically significant
when compared with acetone treated control-1. The average fecundity
of the females that emerged from the treated 4*'' instar nymphs
-39-
§tpsu&s andOSservatUm
decreased linearly with increasing sublethal concentration (y = -1074.3x
+ 200.3, R = 0.842) showing negative correlation coefficient while the
fertility of the egg laid by these affected females also decreased linearly
(y = -1219.x + 184.1, R = 0.878) and showed negative correlation
coefficient (Graph 22 and 23).
Effect on .Longevity
The longevity at different life stages has been investigated with
sublethal concentration of spinosad viz., 0.02%, 0.04%, 0.06%, 0.08%
and 0.1%. The survival duration of 4** instar nymphs was recorded as
5.50 ± 0.28 (t = 0.975, P > 0.05), 6.00 ± 00 (t = 2.400, P > 0.05), 6.50 ±
0.28 (t = 2.182, P > 0.05), 7.00 ± 00 (t = 3.818, P > 0.05) and 7.25 ±
0.24 (t = 2.857, P > 0.05) days respectively. Here the results at 0.02%,
0.04% and 0.06% sublethal concentrations came to be statistically
insignificant while at higher sublethal concentrations at 0.1% and 0.08%
results came to be significant. The 5** instar nymphal longevity was
recorded as 7.25 ± 0.24 (t = 1.428, P > 0.05), 7.75± 0.24 (t = 1.020, P >
0.05), 8.25 ± 0.24 (t = 2.474, P > 0.05), 8.75 ± 0.24 (t = 2.857, P > 0.05)
and 9.50 ± 0.28 (t = 3.236, P > 0.05) days respectively. The results at
lower concentration i.e. 0.02%, 0.04% came to be insignificant while at
higher concentrations i.e. 0.06%, 0.08% and 0.1% results came "to be
statistically significant. The longevity of adult males were 21.50 ± 0.28 (t
= 2.182, P > 0.05), 22.25 ± 0.24 (t = 2.857, P > 0.05), 23.00 ± 00 (t =
-40-
^fsuCts atutOBsermtum
4.787, P > 0.05), 23.75 ± 0.24 (t = 3.780, P > 0.05) and 24.50 ± 0.28 (t =
4.023, P > 0.05) days respectively. The results were statistically
significant at 0.1%, 0.08%, 0.06% and 0.04% while at 0.02% remain
insignificant. The longevity of adult females were also recorded at
aforesaid sublethal concentrations as 17.50 ± 0.28 (t = 1.690, P > 0.05),
18.25 ± 0.24 (t = 2.474, P > 0.05), 19.00 ± 00 (t = 4.330, P > 0.05),
19.75 ± 0.24 (t = 3.500, P > 0.05) and 20.50 ± 0.28 (t = 3.780, P > 0.05)
days respectively. The results at 0.02% came to be insignificant while at
higher concentrations i.e. 0.04%, 0.06%, 0.08% and 0.1% results came
to be statistically significant. The longevity of 4** instar (y = 21.42x +
5.178, R2'= 0.989), 5* instar (y = 26.78x + 6.702, R = 0.994), adult
male (y = 41.07x + 20.48, R = 0.99) and adult female (y =37.5x +
16.75, R = 0.99) of the treated 4* instar nymphs increased linearly with
increasing sublethal concentration. Thus the result showed positive
correlation coefficient (Graph 24-27).
Efficacy of spinosad at 25±1''C temperature, 12L: 12D photoperiod
and 60-70%RH
The initial nymphal mortality upto 24 hours, mortality upto adult
emergence, malformation, fecundity, fertility and longevity were
recorded at 25±1°C temperature, 12L:12D photoperiod and 60-70%
relative humidity. The initial mortality of nymphs upto 24 hours and total
mortality upto adult emergence and malformation were not found during
-41-
^fsu^ muCOBservatum
different stages of life cycle at 25±1°C temperature, 12L:12D
photoperiod and 60-70% relative humidity. The fecundity of females,
which emerged from acetone treated nymphs and untreated nymphs
were found to be as 207.50 ± 5.48 and 213.00 ± 4.45 (t = 1.052, P >
0.05) respectively. The fertility of females, which emerged from acetone
treated nymphs and untreated nymphs were recorded as 184.75 ±1.18
and 195.25 ± 4.21 (t = 1.973, P >0.05) respectively. No statistically
significant effects were shown. The longevity of different life stage i.e.
4*^ 5*^ adult male and adult female were as 6.50 ± 0.28, 6.50 ± 0.28,
22.25 ± 0.24 and 17.50 ± 0.28 respectively. The longevity of untreated
nymphs remains unchanged when compared with untreated nymphs
(control-ll).
Effect on Mortality
The initial mortality of nymphs recorded from the treated 4** instar
at sublethal concentrations viz., 0.02%, 0.04%, 0.06%, 0.08% and 0.1%
were as, 0.33 ± 0.33 (t = 1.736, P > 0.05), 0.67 ± 0.33 (t = 1.877, P >
0.05), 1.33 ± 0.33 (t = 2.645, P > 0.05), 3.00 ± 00 (t = «>, P > 0.05) and
4.33 ± 0.33 (t = 4.773, P > 0.05) respectively. The results at lower
concentrations i.e., 0.02% and 0.04% were statistically insignificant
whereas at higher concentrations i.e. 0.06%, 0.08% and 0.1% found to
be statistically significant. The mortality upto adult emergence were also
recorded at sublethal concentrations as, 0.66 ± 0.33 (t = 1.878, P >
-42-
^^psu&s amCOSsavation
0.05), 3.00 ± 0.57 (t = 3.000, P > 0.05), 4.33 ± 0.33 (t = 4.773, P >
0.05), 7.00 ± 0.57 (t = 4.582, P > 0.05) and 9.33 ± 0.33 (t = 7.007, P >
0.05) respectively. The results were Insignificant at 0.02% while at
higher concentrations results were significant as compared with control-
1. The Initial mortality upto 24 hours and total mortality upto adult
emergence increased linearly with increasing sublethal concentration (y
= 43.31X - 0.5557, R^ = 0.904, y = 95.71x - 0.732, R^ = 0.976
respectively) showing positive correlation coefficient. (Graph 28 and
29).
Effect on Malformation
The malformation in treated 4** instar nymphs at sublethal
concentrations viz., 0.02%, 0.04%, 0.06%, 0.08% and 0.1% were 4.66 ±
0.33 (t = 4.952, P > 0.05), 5.33 ± 0.33 (t = 3.000, P > 0.05), 5.66 ± 0.33
(t = 3.872, P > 0.05), 6.33 ± 0.33 (t = 5.771, P > 0.05) and 6.33 ± 0.66 (t
= 4.063, P > 0.05) respectively (Plate-4). The result shows significant
result as compared to control. The malformation increased linearly with
Increasing sublethal concentrations (y = 52.84x + 2.076, R^ = 0.680)
showing positive correlation coefficient (Graph 30).
Effect on Fecund/ty and Fertility
The fecundity of females, which emerged from the treated 4*
instar nymphs at sublethal concentrations of spinosad viz., 0.02%,
-43-
J^^'Wrf-Qfen^t^, ' "''"'''' I'll'' " ' " , u.uQVo and n 10/ . —
^•°^^. - 0.05) .espec..e,v r.e . s ' « ' ' ' " ' " ' • ' ' ^ ^'^ , .
=°ncen,rat,on were recorded ' " " ' ' ' ^"'"^•^a' recorded as, 140.75 + Ofl"; »
' ' ^ • = 0 - . a 4 ( , = a . 8 4 , P > 0 0 5 ) S S 7 / " ' ' ^ ' ' ^ ° ° ^ ' '
^ '-^-—- J : :;;:::: - °- '= - ^ compared wl,h contro, , JU ' "''"'""'"' * " « "
' ' -eased „near,v mu i n c r e ' " " ' ' ' " " " * ' ° ' '^^ ^ - ^ ' -
"^•3. R = 0.978) Showing neaat,v» "
(^-P-^ 31), White the fertility of tH " ' " " ' ° " ~ ^ ' " * " ' 3 , . , . , _ ^ ' - e . , . , d . , t h e 3 . e c , e d f e . a i e s
-native correlation coefficient (Graph 32, " " ' ' ' ^ ='°^^"^
Effect on Longewty
' " ^ ^ ' " ' ° " '°"9-^'V were investigated t>v . "ymP' s Of Oy . .e . . s fee.,, , , „ . , ' ' ' ^ ' " * « ^'^ - t a r
°----.oe.,ooarr"""""°"^^"' *"-^ ' ^-^o/o, and 0 1% r,f
""nation Of treated 4%nstarn. K ' " " ° ' ' * " ^ ="™Val '"Star nymphs werp « 7c
°0^X 7.25 . 0.24 (t - , eao P n ' " ' ^ ' = "'^^^^ ^ > V / .oyu, P > 0 0 5 ) 7 7C
°°5) ,8 .00t0O(t = 3 2 7 3 p ^ ' ' ' * "'^^ « = 2-182, P >
^•273, P> 0.05) and 8 .50.0 28 rt , - "ZS (t = 2.672, P > -44-
^S^fSuO^ atufOBservatum
0.05) days respectively. The results were significant only at two higher
concentration i. e. 0.1% and 0.08% while, rest of sublethal
concentration shows statistically insignificant effects. The longevity of 5*
instar nymphs recorded as 7.25 ± 0.24 (t = 1.690, P > 0.05), 8.00 ± 00 {t
= 3.273, P >0.05), 8.50 ± 0.28 (t = 2.672, P > 0.05), 9.00± 00 (t = 4.226,
P > 0.05) and 9.25 ± 0.24 (t = 3.237, P > 0.05) days respectively. The
results gave statistically significant at higher concentration
{0.1%,0.08%, 0.06%, 0.04%) while at 0.02%. results was insignificant
effects. The male longevity recorded as 23.50 ± 0.28 (t = 2.182, P >
0.05), 24.00 ± 00 (t = 3.818, P > 0.05), 24.75 ± 0.24 (t = 3.194, P >
0.05), 25.25 ± 0.24 (t = 3.499, P > 0.05) and 26.25 ± 0.24 (t = 4.040, P >
0.05) days respectively. The results are insignificant only at 0.02% while
rest of all higher sublethal concentration remain significant. The female
longevity recorded as 17.75 ± 0.24 (t = 0.976, P > 0.05), 18.25 ± 0.24 (t
= 1.690, P > 0.05), 19.00 ± 00 (t = 3.273, P >0.05), 19.75 ± 0.24 (t =
2.927, P > 0.05) and 20.25 ± 0.24 (t = 3.236, P > 0.05) days
respectively. Here, the results at higher concentrations (0.1%, 0.08%,
0.06% is significant while rest of sublethal concentrations remained
insignificant. The affected longevity of 4* instar nymphs (y
=20.35x+6.440, R = 0.991), 5* instar nymphs (y =27.85x+6.690,. R =
0.972), adult males (y =37.14x+22.47, R = 0.982) and adult females
-45-
^^fisu&s (md^OSservatmi
(y =29.28x+17.28, R = 0.980) increased linearly with increasing
concentration of spinosad (Graph 33-36).
-46-
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H 6 -i
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Showing initial mortality upto 24 hours at 30±1°C/14L:10D
y=1004.4x+1.9103
R* = 0.8785 ^ ^ / ^ ^
^ ^ • 30±1X/14L:10D
^ ^ ^ ^ Linear (30+1X/14L:10D)
1 ! [ !
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 1. Fitted linear regression line for mortality upto 24 hours of treated 4* instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperlod.
Showing total mortality upto adult emergence at
4>
g 18 o) 16
^ 14 ^
t 12 5 10 •!»</> 8 -i o o Q.Z 6 3 ^
>» 4 i . 1 2 f o 0 4 -E « 0
o 1-
30±rC/14L:10D
y= 1624.7x4 2.3871 R^= 0.9181 y^^
• ^/ '^ ^ ^ • 30+1"C/14L:100
/ ^ Linear {30±1°C/14L:10D)
.,_ , ,j , , , .
0.002 0,004 0.006 0,008 0.01
Concentration %
Graph: 2. Fitted linear regression line for mortality upto adult emergence of treated 4* ^ instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1 "0/141:100 temperature and photoperlod. ,-.^-
Showing malformation at30±1*C/14L:10D
Vt i
o ^ 4 \
i • ^ 2
V= 417.14x+1.5992 R = 0.6657
• 30±1''C/14L:10D
- Linear (30±1''C/14L:10D)
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 3. Fitted linear regression line for malformation of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperiod.
Showing fecundity at 30±1X/14L:10D
y=-15864x4 192,6
R = 0.8551
• 30±1"C/14L:10D
Linear (30±1X/14L:10D)
0.002 0,004 0.006 0,008 0.01
Concentration %
Graph: 4. Fitted linear regression line for fecundity of adult female, which emerged from treated 4* ^ instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperiod.
-56-
Showing fertility at 30±rc/14L:10D
250
200
w
y--17018x4 172.3 R = 0.8583
I 150 f
Ig 100 a>
• 30±1''C/14L:10D
Linear (30±1°C/14L:10D)
0 0.002 0.004 0.006 0,008 0.01
Concentration %
Graph: 5. Fitted linear regression line for fertility of adult female, which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperiod.
Showing longevity of treated IV**' instar at 30±rc/14L:10D
(0
I c o
8
7
6
5
4
3
2
1
0 - i -
• 30±1'C/14L:10D 7=176.32x4 5.3677
R = 0,9764 Linear (30±1°C/14L:10D)
0 0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 6. Fitted linear regression line for longevity of treated 4"" instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperiod.
-57-
Showing longevity of V**' Instar nymph which emerged from treated IVth Instar at
30±rC/14L:10D
> OJ c o
• 30+1'C/14L:100
V-223.54x^ 7.0845 R = 0.929 - Linear
(30±1X/14L:100)
0.002 0.004 0.006 0.008
Concentration %
0.01
Graph: 7. Fitted linear regression line for longevity of V** instar which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±rC/14L;10D temperature and photoperiod.
>> (0
>. • >
a> O)
o - J
Showing longevity of adult male which <
24.5 24
23.5 23
22.5 22
21.5 21
20,5
20
emerged from treatedIV*''instarnymphs at30±l''C/14L:10D
i H J
i
1
1
^ _
0
A / ^ ^ y= 387.99x+21.075 ^ . ^ R = 0.872
0.002 0,004 0.006 0.008 0.01
Concentration %
• 30±1"C/14L:10D
Linear (30±1"C/14L:10D)
Graph: 8. Fitted linear regression line for longevity of adult male which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±rC/14L:10D temperature and photoperiod.
-58-
Showing longevity of adult female which emerged from treated IV'^instar nymphs at
30±rC/14L:10D 25
> (D O) c o
10 y=487.0xt 16.699
R = 0.9676 • Linear (30±1"C/14L:10D)
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 9. Fitted linear regression line for longevity of adult female which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 30±1°C/14L:10D temperature and photoperiod.
(A
z 5 8 o
o 5
1 - 5 2» 4
t : 3
i 2 S 1 £ 0
T
-
4 >
. ^_,_
0
Showing initial mortality upto 24 hours at
r—
0.002
25±rC/12L:12D
V= 727.14x4 1 . 3 4 M , ^
R = o.soy^ ^ ^ ^
^ ^ • 25+lX/12L:12D
Linear (25+rC/12L:12D)
J , .—-r~----- — ;
0.004 0.006 0.008 0.01
Concentration %
Graph: 10. Fitted linear regression line for mortality upto 24 hours of treated 4* instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
-59-
u c 4)
Showing total mortality upto adult emergence at 25±rC/12L:12D
y=1364.5x+ 2.3171 R = 0.9048
• 25+lX/12L:12D
• Linear (25±rC/12L:12D)
0.002 0.004 0.006 0.008
Concentration %
0.01
Graph: 11. Fitted linear regression line for mortality upto adult emergence of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±rC/12L:12D temperature and photoperiod.
Showing malformation at 25±1*C /12L: i2D
v> o Z c o m E
m
• 25±1'C/12L:12D
V= 485.73x4 0.6495 R = 0.9127 - Linear
(25±1°C/12L:12D)
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 12. Fitted linear regression line for malformation of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
-60-
Showing fecundity of females at 25±1 "0/121:12D
250
y=-15623x+ 174.59 R2-0.8681
• ZStlX/lZL-.lZD
Linear (25+l*C/12L:12D)
0 0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 13. Fitted linear regression line for fecundity of adult female, which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
1 2' 1 41
200 180
160 140
120 100 80
60
40
20
0
Showing fertility of female at 25±1
t - i V ,
1
( i
J
0
• ^
0.002
y=-15827x+150.6 R - 0.8638
• \ ^
1 1 1
0.004 0.006 0.008
Concentration %
0.01
"C/12L:12D
• 25±1'C/12L:12D
Linear (25±1''C/12L:12D)
Graph: 14. Fitted linear regression line for fertility of adult female, which emerged from treated 4*" instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
-61-
Showing longevity of IVth Instar nymph at 25±rC/12L:12D
(A >» (0
> a> o> c o
y=214.05x+6.6163 R = 0.9817
• 25±rC/12L:12D
Linear
(25±1°C/12L:12D)
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 15. Fitted linear regression line for longevity of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
Showing longevity o f Vth instar at 25±1°C/12L:12D
• 25±l''C/12L:12D
V-339.82X+5.8616 R2 = 0.9542 Linear
(25+rC/12L:12D)
0 4
0 0.002 0.004 0.006 0.008
Concentration %
0.01
Graph: 16. Fitted linear regression line for longevity of 5^^ instar which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±1°C/12L:12D temperature and photoperiod.
-62-
Showing longevity of adult male at 25±1°C/12L:12D
27 -
• 25+l'-C/12L:12D
y=456.81x t 23.011 R = 0.9216 - Linear
{25±1°C/12L:12D)
0.002 0.004 0.006 0.008 0.01
Concentration %
Graph: 17. Fitted linear regression line for longevity of adult male which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±rC/12L.12D temperature and photoperiod.
in >. (0
t>
c o
20
19.5
19 -
18.5 -
18
17.5 i
17
C
^
)
Showing longevity of adult female at 25±rc/12L:12D
0.002
^4f • 25±1''C/12L:12D
y=236.12x» 17.584 Lmp^^
R' = 0-9852 (25±rC/12L:12D)
1 \ i n
0.004 0.006 0.008 0.01
Concentration %
Graph: 18. Fitted linear regression line for longevity of adult female which emerged from treated 4'^ instar nymphs of D. koenigii with various sublethal concentrations of imidacloprid at 25±rC/12L:12D temperature and photoperiod.
-63-
in o
3 O
O 'a. Zi
*^ 15 t o £ « 'c
10
8
6
4
2
0
-2
Showing Initial mortality upto 24 hour at 30±rC/14L:10D
y=91.9x-0.6S33 R2 = 0.973
• 30+l'C/14L:10D
• Linear (SO+l-C/lAL-.lOD)
0.02 0.04 0.06 0.08
Concentration %
0.1
Graph: 19. Fitted linear regression line for mortality upto 24 hours of treated 4'^ instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1°C/14L:10D temperature and photoperiod.
4> O
%^2 H
« ^ 0 1 o ±-U 8 ^ 2^ * ^ (TJ 4
1 2 0 ^
-2 6
Showing mortality upto adult emergence at 30±rC/14L:10D
^y^ y^ y = 176.67X - 0,55*6 30±rC/14L-.10D
y ^ R - 0.9876
y^ Linear y ^ (30+l"C/14L:10D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 20. Fitted linear regression line for mortality upto adult emergence of treated 4'" instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±rC/14L:10D temperature and photoperiod.
-64-
Showing malformation at 30±1'C/14L:10D
• 30±l''C/14L:10D
y=31.857x+2.7938 R = 0.2872 • Linear
(30±1"C/14L:10D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 21. Fitted linear regression line for malformation of treated 4*^ instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1 "0/141:100 temperature and photoperiod
250
Showing fecundity of adult female which emerged from treated IV** instar nymph at
30±rC/14L:10D y -1074.3X t 200.3
o Z >% •5 c O 4)
200
150 -
100
50
0 H
c
>-., . ,^^^ R' = 0.842
1 1 ! '•
) 0.02 0.04 0.06 0.08
Concentration %
- ^
0.1
• 30±1'"C/14L:10D
Linear (30±1'C/14L:10D)
Graph: 22. Fitted linear regression line for fecundity of adult female, which emerged from treated 4*" instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1°C/14L;10D temperature and photoperiod.
-65-
Showing fertility of female emerged from treated IV i lnstar nymph at 30±1°C/14L:10D
250
_ 200 V=-1219.5xf 184.19 R = 0.8785
• 30±1'"C/14L:10D
Linear (30+l"C/14L:10D)
0.02 0.04 0.06 0.08
Concentration %
0.1
Graph: 23. Fitted linear regression line for fertility of adult female, which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1°C/14L:10D temperature and photoperlod.
Showing longevity of treated IV*" instar at 30±rC/14L:10D
m
I c o
V= 21.429x4 5.1786 R = 0.989
• 30±1''C/14L:10D
• Linear {30±1°C/14L:10D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 24. Fitted linear regression line for longevity of treated 4" instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±rC/14L:10D temperature and photoperiod
-66-
Showing longevity of V*'' instar emerged from treated IV*" instar at 30±1°C/14L:10D
• 30±1"C/14L:10D y^ 26.786x4 6.7024
R'= 0.9941 2 J Linear
(30±1'-C/14L:10D)
0.02 0.04 0.06 0.08
Concentration %
0.1
Graph: 25. Fitted linear regression line for longevity of 5*' instar which emerged from treated 4* instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1°C/14L:10D temperature and photoperiod
Showing longevity of adult male emerged from trated IV** instar nymphs at 30±1°C/14L:10D
a •D
> O) c o
30 1
25 J
20
15 i
10 ;
0 4
• 30+rC/14L:10D
y^41.071x+20.488 R = 0.99 • Linear
(30±1''C/14L:10D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 26. Fitted linear regression line for longevity of adult male which emerged from treated 4* " instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±rC/14L:10D temperature and photoperiod.
-67-
Showing longevity of adult female emerged fron treated IV*' instar nymph at 30±1°C/14L:10D
V=37,5x > 16.75
R ^ - 1
• 30±1X/14L:10D
• Linear (30±1''C/14L:10D)
0 ^
0 0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 27. Fitted linear regression line for longevity of adult female which emerged from treated 4** instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 30±1°C/14L:10D temperature and photoperiod.
>" in O Z
3 o XL
O Q. 3 >» 15 •c £
.2 c
5
4
3
2
1
0
-1
I
1 i
1
, i
' i !
• • 1 ,
• -
Showing initial mortality upto 24 hours at
0.02
^ ^
0.04
25±rC/12L:12D
•
^ ^ ^ • 25±l''C/12L:12D ^ ^ y-43,314x-0.5557
^ ^ ^ ?} -~ 0.9043 • Linear
(25+rC/12L:12D)
0.06 0.08 0.1
Concentration %
Graph: 28. Fitted linear regression line for mortality upto 24 hours of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
-68-
Showing total mortality upto adult emergence at 25±rC/12L:12D
• 25±1'C/12L:12D
y^95.714x-0.7324 R = 0.9767
0.02 0.04 0.06 0.08 0.1
Concentration %
Linear (25±1X/12L:12D)
Graph: 29. Fitted linear regression line for mortality upto adult emergence of treated 4** instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
I I a
• 25±1''C/12L:12D
Linear (25+lX/12L:12D)
Concentration %
Graph: 30. Fitted linear regression line for malformation of treated 4' instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1 "0/121:120 temperature and photoperiod.
-69-
w o z >> *•* c 3 u
250
200
150
100
50
n
1
4 ( i J 1 j
0
Showing fecundity of adult female at 25±1°C/12L:12D
0.02
y=-1245.7x+197.37 R' = 0.9781
0.04 0.06 0.08 0.1
Concentration %
• 25±l'-C/12L:12D
Linear (25±1''C/12L:12D)
Graph: 31. Fitted linear regression line for fecundity of adult female, which emerged from treated 4" instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
200
- r 150
o Z ^ 100 -•E <v u. 50
0
(
Showing fertility of adult female at 25±rC/12L:12D
--v^,^^ V=-1288.6x+172.93 ^•"^^-^.^^^ R = 0.9723
^ " ~ \ , . ^ ^ • 25±1*C/12L:12D
^ ^ ^ Linear {25+l"C/12L:12D)
1 1 ! • • r 1
) 0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 32. Fitted linear regression line for fertility of adult female, which emerged from treated 4'* instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
-70-
n
I at c o
Showing longevity of IV** instar nymphs at 25±rC/12L:12D
y= 20.357x4 6.4405 R = 0.9911
• 25±1'C/12L:12D
Linear (25±1°C/12L:12D)
0.02 0.04 0.06 0.08
Concentration %
0.1
th ;. Graph: 33. Fitted linear regression line for longevity of treated 4 instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
(0 n •a
l c o
Showing longevity of V** Instar nymphs at 25±rC/12L:12D
10
8
6 j
4 ] I
2 H
0 +-
y=27.857x+6.6905 R = 0.9729
0.02 0.04 0.06 0.08
Concentration %
0.1
• 25+l'C/12L:12D
Linear (25±1"C/12L:12D)
Graph: 34. Fitted linear regression line for longevity of 5^*" instar which emerged from treated 4* ^ instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±rC/12L:12D temperature and photoperiod.
-71-
Showing longevity of adult male at 25±rC/12L:12D
o-i ,„o -n All 25±rC/12L:12D y = 37.143x+22.4/6 R = 0.9821
Linear (25±1"C/12L:12D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 35. Fitted linear regression line for longevity of adult male which emerged from treated 4* instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±1°C/12L:12D temperature and photoperiod.
Showing longevity of adult female at 25±rC/12L:12D
• 25±1X/12L:12D
y= 29.286x4 17.286 Lj^ear R = 0.9802 (25±rC/12L:12D)
0.02 0.04 0.06 0.08 0.1
Concentration %
Graph: 36. Fitted linear regression line for longevity of adult female which emerged from treated 4'*" instar nymphs of D. koenigii with various sublethal concentrations of spinosad at 25±rC/12L:12D temperature and photoperiod.
-72-
Plate 1: Malformation In adults emerged from 4* instar treated nymphs following topical application of imidacloprid at 30±rc/14L:10D temperature and photoperiod.
Plate 2: Malformation in adults by treated 4* instar treated nymphsJoll^^g topical application of imidacloprid at 25±rC/12L:12D temperature awtJphotoperi J
. /
2Ww L//1??]
M
Plate 3: Malformation in adults emerged from 4**" instar treated nymphs following topical application of spinosad at 30±1°C/14L:10D temperature and photoperiod.
Plate 4: Malformation in adults emerged from 4'^ instar treated nymphs following topical application of spinosad at 25±1°C/12L:12D temperature and photoperiod.
- 7 4 -
m M
^iieajuuefi
r [f-
DISCUSSION
Application of chemical insecticides is still one of the most
powerful measures against insect pests. Insecticides are often the only
effective remedy for quickly and inexpensively reducing pest
populations to below economic injury levels. The use of insecticides is
generally the preferred strategy for their high efficacy during a. pest
outbreak. However, there are many negative side effects associated
with the frequent spraying of chemical compounds, especially to human
and environment. There have been efforts, during the past few
decades, to develop biorational insecticides and systemic
neonicotinoids affecting the target pest with minimum effect on natural
enemies and the environment. This approach has led to the
development of botanicals (such as citrus oil derivatives, neem-
azadirechtin, garlic oils, hot pepper oils etc), soaps and oils, microbial
insecticides (like Bacillus thuringiensis, Beauveria, Steinernema),
growth regulators, pheromones, non-organic natural products,
fermentation products (like spinosads and avermectins) and
neonicotinoids (like acetamiprid, clothianidin, dinotefuran, imidacloprid,
nitenpyram, thiacloprid, thiamethoxam etc), which are able to efficiently
control agricultural pest species with minimum effects on natural
enemies. Spinosad is a mixture of spinosyns A and D, which are
fermentation products of the soil actinomycete Saccharopolyspora
(Oiscussion
spinosa (Mertz and Yae 1990, and Thompson et al., 1997). It is a
broad-spectrum natural bio-insecticide offered a new mode of action
which is relatively safe on natural enemies (Temerak 2003). This
compound has two unique modes of action, acting primarily on the
insects nervous system at the nicotinic acetylcholine receptor, and
exhibiting activity at GABA receptor (Salgado 1997, Watson 2001).
Imidacloprid which belongs to the class neonicotinoid group is similar to
the natural insecticide nicotine and acts on the central nervous system.
It blocks a specific neuron pathway that is more abundant in insects
than warm-blooded animals, hence this insecticide is more selectively
toxic to insects than mammals. In animals and humans, imidacloprid is
quickly and almost completely absorbed from gastrointestinal tract, and
eliminated via urine and feces within 48 hrs.
The newer insecticides are not immune to the development of
resistance in the insect pests, and since their registration, several cases
of resistance have been reported (Horowitz et al., 2002, 2004, 2007).
However, the availability of compounds with diverse modes of action
has assisted in managing resistance to insect pests. As most of the new
biorational insecticides and neonicotinoids have shown effectiveness
against insect strains having resistance to conventional insecticides with
no evidence of cross resistance, they can play an important role in IPM
programs.
-76-
(Discussiott
It is a well known fact that the efficacy of insecticides is greatly
influenced by several biotic and abiotic factors such as target species,
temperature, humidity, photoperiod, type of surface that insecticide is
applied to. The effect of temperature on the efficacy can either be
positive or negative. The response relationship between temperature
and efficacy has been found to vary depending on the mode of action of
the insecticide, target species, method of application and quantity of
insecticide ingested or contacted (Johnson, 1990). Stability,
vaporization, penetration, activity and degradation of insecticides are all
partly dependent on physical and biochemical processes that proceed
at characteristic rates at different temperatures. Positive correlations of
temperature with effectiveness for organophosphorus insecticide have
been noted in the laboratory tests by Zwick (1962) using malathion at
21 and 25.5°C against Sitophilus granarius. Also using malathion,
Teotia and Pandey (1967) reported the positive correlation against 7.
casteneum at 27°C and 35°C. O'Donnell (1980) found positive
correlations between temperature and effectiveness of insecticide in T.
confusum with malathion, jodfenpho, pirimiphosmethyl and fenitrothion.
The temperature may synergize activity of an insecticide, or an
insecticide may enhance the impact of high temperature. Ebeling (1990)
demonstrated this by exposing confused flour beetles, T. confusum, and
German cockroaches, Blattella germanica to bo r i c ^ i t f ^ ^ ^^ f t a t 46°C;
-77-
Oiscussian
the mortality due to heat and boric acid together was greater than the
due to either factor alone. Fang et al., 2002a; Subramanyam et al.,
2003; Toews and Subramanyam 2003; Toews et al., 2003; Nayak et al.,
2005; Daglish and Nayak 2006; Getchell 2006; Subramanyam 2006;
Subramanyam et al., 2007 also observed that the efficacy of spinosad is
affected by several biotic or abiotic factors, such as the target species,
the type of commodity, the exposure interval and the type of surface
that spinosad is applied to. These parameters are crucial for the
performance of the currently used grain protectants, since they
determine their efficacy (Noble e^ al., 1982; Johnson 1990; Braness et
al., 1998; Arthur 1996, 1999). For example, pyrethroids are generally
less effective at high temperatures, while the reverse is true for OPs
(Johnson 1990). Musser and Shelton (2005) found that the increase in
temperature decreased the efficacy of spinosad against Ostrinia
nubilalis (Hubner) (Lepidoptera: Crambidae). However, there is still
inadequate information on the effect of temperature and humidity on the
insecticidal effect of spinosad and imidacloprid against hemipterous
insect pests. Combining or integrating temperature with other control
strategies would be beneficial for modern pest management
approaches for controlling insect pest (Arthur and Dowdy 2003). In the
present work, we examined the efficacy of imidacloprid and spinosad
against Dysdercus koenigii, (Hemiptera: Pyrrhororidae) at two different
-78-
(Discussion
temperatures and photoperiods i.e. 30±1°C/14L:10D and
25±1 °C/12L: 12D with constant 60-70% relative humidity.
Imidacloprid
In the present investigation topical application of sublethal
concentrations of imidacloprid viz., 0.008%, 0.006%, 0.004%, 0.002%,
and 0.001% on 4* Instar nymphs of D. koenigii at tvi/o different
temperature and photoperiod (i.e. 30+1 °C/ 14L:10D and
25±1°C/12L:12D) with constant relative humidity (60-70%) caused
immediate mortality within 24 hrs and successive mortality upto adult
emergence. This initial mortality within 24 hour as well as mortality upto
adult emergence increased with increasing sublethal concentrations at
both the temperature and photoperiod. The results are similar to those
reported by Eger e^ a/., (1998) in which the mortality of Franklinella
species showed concentration dependent response. Tows and
Subramanyum (2003) also reported the mortality of T. casteneum that
increased with increasing dosage of insecticide. The mortality within 24
hour as well as total nymphal death upto adult emergence was
comparatively higher at 30±rC/14L: 10D as compared to 26±1°C/12L:
12D temperature and photoperiod with constant (60-70%) relative
humidity. The total nymphal loss within 24 hour after the topical
application of highest selected sublethal concentration of imidacloprid
(i.e. 0.008%) at 30±rc/14L: 10D was 30%, whereas, it was 6.67 %
-79-
(Discussion
less when the treatment was made at 25±rC/12L: 12D. However, the
topical application of the lowest selected sublethal concentration (i.e.
0.001%) resulted in 11.1% and 7.76% nymphal death at 30±1°C/
14L:10D and 25±1°C/12L:12D temperature/photoperiod respectively as
compared to control. The total nymphal loss upto adult emergences
following the topical application of 0.008% imidacloprid at 30±1°C/ 14L:
10D and 25±1°C/ 12L: 12D temperature and photoperiod was 46.66 and
40.00% respectively. Agarwal and Tilak (2006) studied the efficacy of
imidacloprid gel bait (2.15%) upto 8 weeks against German cockroach
and reported that there was 96.8 to 98.8% reduction in the population
as compared to control. Pruthi and Jerath (1960) also determined the
effectiveness of endrin against Dysdercus cingulatus and Chrotogonous
trachypterus adults at different temperature and found that the toxicity is
accelerated with rise in temperature. Uddin and Ara (2006) studied the
toxicity of six insecticide on Trilobium casteneum (Herbst) at the
temperatures of 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30 and 32°C and
observed that there was a marked decrease in the effectiveness of all
insecticide tested with a lowering of temperature. The order of toxicity of
insecticide was chloropyrifos > lamda-cyhalothrin > Malathion >
cypermethrin > carbosulfan > dicofol. Ming and Qiang (2002) studied
the selective toxicity of six different insecticides against Myzus persicae
and receded the order of toxicity as imidacloprid + SV1 > imidacloprid >
-80-
(Discussion
endosulfan > methomyl > fanvalerate > dimethoate. Ateyyat et. al.,
(2009) compared the toxicity of aqueous extracts of nine plants with
imidacloprid against sweet potato whitefly, Bemissia tabaci and found to
killed 71% of early stage nymphs, which was not significantly different
from mortality caused by imidacloprid. Khowaja et. al., (1995) reported
that the topical application of 10, 08, and 06 ppm of temik (aldicar) to
the 4'* instar nymphs of Dysdercus cingulatus caused 50, 41, and 27%
more nymphal loss than that of control 2% respectively. The initial
mortality following the topical application of higher sublethal
concentrations (i.e. 80 and 60 ppm) of cythion on 4^^ and 5" instar
nymphs of D. Cingulatus was very high within 24 hrs, however, the
subsequent mortalities up to adult emergence were insignificant. The
total nymphal loss and survival duration of the treated insects were
directly proportional and increased linearly with the increase in
concentration, whereas, the fecundity & fertility of the adults were
inversely proportional and decreased linearly with increase in
concentrations (Khowaja et. al., 2001 and Khowaja and Qamar, 2005).
The malformation, moult inhibition and emergence of abnormal/
sub-abnormal individual increased with increasing sublethal
concentration of imidacloprid at these two temperatures and
photoperiods (Plate-1 & 2). The percentage of malformation at the
highest selected sublethal concentration, i.e. 0.008%, was 16.67 and
-81-
Oiscussion
14.43% respectively, whereas, at lowest concentration (i.e. 0.001%) the
percent malformation was 8.86 and 2.23% respectively. Tiwari et. al.,
(2006) noted that the topical application of neem based insecticides on
Dysdercus koenigii caused ecdysial stasis and development of
adultoids and imagoes with varied degree of deformalities. Topical
application of azadiractin through contact method evoked various
specific and non-specific effects during the course of development in
various stages of red cotton bugs (Koul, 1984). RD-9 Replin at higher
concentration prolonged the nymphal period and affected the
emergence of adults of Dysdercus koenigii (Gupta et. al., 1997).
Khowaja et. al., (1995) reported that the topical application of temik
(aldicarb) to the 4* and S**" instar nymphs of D. Cingulatus at 10, 08, and
06 ppm dose produced 7.1, 6.3, 4.8% and 9.6, 8.2, 7.0% adults with
malformed wings respectively. In contrary to the earlier reports some
insecticide, such as imidacloprid and fanvalerate, increased the
percentage of alate offspring of aphids. Field observations of the cotton
aphid, Aphis gossypii, by Kring and Conway (2004) also indicated an
increase in the percentage of winged aphids in cotton fields after an
application of imidacloprid. Many factors such as crowding (Kawada
1989; Matsumura 1996), host plant quality (Lees 1966; Lu and Chen
1993), photoperiod (Joknson 1966; Uddin and Baqui 1993),
temperature (Liu and Wu 1994; Joknson 1966), hormones (Zera and
-82-
discussion
Tobe 1990; Fairbairn 1994) and seasonal changes (Muraji et. al., 1989)
could affect aphid wing development.
The fecundity and fertility of D. koenigii following the topical
application of different sublethal concentrations of imidacloprid at
30±1°C/14L: 10D and 25±1°C/12L: 12D temperature/photoperiod with
constant relative humidity were also adversely affected. The reduction
in egg production and egg hatching of the affected females emerged
from the 4* instar treated nymphs were concentration based and
increased with increasing sublethal concentration. On comparing the fall
in egg production, it was more pronounced at 25±1°C/12L: 12D as
compared to 30±1°C/14L: 10D. Following the topical application of the
highest selected sublethal concentration (i.e. 0.008%) at 30±1°C/14L:
10D, the fall in average egg production was 63.28%, whereas, this loss
was enhanced to 5.27% when the treatment was made at 25±1°C/12L:
12D. At lowest selected sublethal concentration, the reduction in
fecundity was 20.72 and 23.97% respectively as compared to control.
Reduced fecundity of D. koenigii affected with sublethal concentrations
of imidacloprid is consistent with previous studies investigating other
hemipterous insects. The fecundities of two aphid species {Myzus
persicae and /W. nicotiana Blackman), two green leafhopper species
{Neptioteitix virescens, Distant and N. cincticeps Uhler) and a brown
planthopper species {Nilaparvata lugens Stal) were reduced by more
-83-
(Discussum
than 50 % after feeding on plants/leaves treated with sublethal
concentrations of imidacloprid (Divine et. al., 1996; Widiarta et. al.
2001; Bao et. al., 2009). Similarly, Lashkari e^ a/., 2007 reported that
the oviposition rate, longevity and growth of cabbage aphid, Brevicoryne
brassicae L., were negatively affected by sublethal concentration of
imidacloprid. Nishi and Takahashi, (2002) observed that the average
numbers of egg laid per ovipositing female of Amphibolous venator
increased with increasing temperature (i.e. 25°C, 27.5°C, 30°C 32.5°C
and 35°C/ 60-70%RH and 16L-8D of photo regime), however
differences were not significant. RH 5849, an analogue of halofenozide,
was shown to inhibit ovariole development in Lepidoptera and some
Coleoptera (Wing e . al., 1988). Monthean and Potter (1992) found that
feeding on foliage treated with RH 5849 (1,2-dibenzoyl-1-
tertbutylhydrazine) reduce the fecundity of Japanese beetles by 66-
74%, but did not reduced the percentage of hatch. Aller and Ramsay
(1988) also observed that ingestion of RH 5849 by newly emerged
adults of several phytophagous beetle species prevented oviposition
due to the absence of developing oocytes, whereas, the ingestion of RH
5849 by older female caused cessation of oviposition due to the
resorption of the oocytes. Destruction of egg chamber after treatment
with tepa, metapa, apholate, hempa, methlmethane sulphonate and
hydroxy urea has been reported by Morgan and La Brecque, (1964);
-84-
Oisotssian
Morgan, (1967); Kissam et. al., (1967). The effects on division of
oogonia in the germahum, the inhibition of growth of oocytes and
trophocytes, degeneration of follicular epithelium and the resorption of
contents of egg chamber has been reported by Rezabova and Landa,
(1967); Landa and Matol, (1971); Khowaja et. al., (2001) and Khowaja
and Qamar, (2005). The application of 06, 04, and 02 pg of 22, 25-
dideoxyecdysone to the 4** instar nymphs of Dysdercus cingulatus
reduced the egg production of the emerged females by 30.9, 18.5, and
11.0% respectively as compared to control, however, the application of
the corresponding doses of 2,22,25 trideoxy-5phydroxyecdysone
decrease the fecundity by 27.6, 21.4, and 15.4% respectively as
compared to control (Khowaja et. al., 1996). The fertility of the females
emerged from the IVth instar treated nymphs following the treatment of
0.008% imidacloprid at 30±1°C/14L: 10D and 25±1°C/12L: 12D reduced
by 75.11 and 78.34% respectively, however, at the lowest concentration
the reduction was 26.42 and 28.28% respectively. It has been noted
that the reduction in fertility was more at 25±1°C/12L: 12D than
30±1°C/14L: 10D temperature and photoperiod. Boina et. al., 2009 also
observed that the fecundity and fertility of the Asian citrus psyllid,
Diaphorina citri, were reduced significantly following the long term
exposure of imidacloprid through systemic method. The topical
application of various sublethal concentrations of monocrotophos to 4 *"
-85-
OiscMSsion
and 5* instar nymphs of D. cingulatus resulted in linear fall in egg
production and egg hatching and showed a negative correlation with the
increasing concentrations (Khowaja et. al., 1994 and 2002). Likewise,
the average fertility of the eggs laid by the affected females of D.
Cingulatus, after the topical application of higher selected doses (06,
04, 02 pg of 22, 25-dideoxyecdysone) on 4'^ instar nymphs, decreased
by 35.3, 30.1, and 26.0% respectively as compared to control, however,
with corresponding dose of 2,22,25trideoxy-53hydroxyecdysone, the
hatchability of the eggs was dropped by 29.2, 20.3, and 13.8%
respectively as compared to control (Khowaja et. al., 1996). Inhibition in
fecundity and fertility after the application of natural ecdysones or their
analogues have also been reported by Jalaja et. at., (1976), Ahmad &
Khan (1985), Ahmad (1983), and Khan et. al., (1990). Wright et. al.,
(1970) observed that p-ecdysone prevented lipid synthesis which is
necessary for vitellogenesis. On the contrary, Zeng and Wang (2010)
reported that the exposure of sublethal dose of imidacloprid to
Tetranychus cinnabarinus led to a significant increase in the hatch rate
of eggs. James and Price (2002) also reported that the twospotted
spider mites, Tetranychus urticae, either directly exposed to spray
formulations or fed on discs cut from a systemically treated bean plant
produced 10-26% more eggs during the 1'* 12day of adult life and 19-
23% more during adult hood compared with a water only treatment.
-86-
"-—««»».,, r:*"'-"*--*«
0008%) , , "^""centratjon im/rt , ^°' ^ ' 30±l.c/i4L- 10D ° " ""'^3<:'0Prid (i.e.
tempera,..e/pho,ope.oc< on 4%n., " " ' ' * ' ° ^ ' ' ^ ^ - ^20
J^'t/b/oancf 3 84% r^. *=" me surv/Va/
^ ' ^ - - n V ' ^ P . s , emerge. . 0 . . " " ' ^ ^ ^ ^ ' ^ ' — • T.e
'^•^2 and 46. ^5% „,,, 3, „ '^^ated nyn^p^s,
^ ° " ^ ^ " " " ° " « - - O a „ . . J •"''^^''^---=°" ' - ' - ^"e ,.e span of ,,e s . ' " ' " " " ' > ' ^^ - -Pared ,0
'"'-^^---.ooa.Ir^^"^^"--"--v - - ^ » / . . . p . . . , , . ^ ^ : : - - - a 3 , . 0 . 0 0 , . . . . . , :
P^ ed to contro/. L/kew/se fh ,
-87-
° ^ ^ ^ - enhance. . , , , 33 ^^^ ^ ^ " " ^ °^ '^^^^^^^^^
— a « o n (,e. o.oo. / , , , J " " ' ' " ' " ^ ' ^ ' - ' ' - a s , a, ,owes, /oy 11 Was onlv 1 AO
^ - ' - ^ results are in agree.e , ' " ' ' " " ' '^^P-'-ely
'°P'=al appncation of highest , , ' ' ' " ' ' " ^ ' '"^ •"ynest selected QI.M *..
'"°"ocro.ophos on 4' instar <=°"oen.ration of mstar nymphs resuiten • .
"V -PM ...a«on, Whereas ,he , ''°''° ' "=^^ - ^^ " «'eas, the longevity of V" ; .
- - pa red focomroi. ^ ' "^"^"^^ ' - ^ 50.9% , , , , ^^
^eng and Wang ^2010^ r.
="«^"V P-onge. an. increasef ^ ^ " ^ ~ ° " -
' ^ - " ' - ' - - - - o f s i ; : : : ^ " ' ' ^ - - - -- - ' ^ e n , e a n , e n g . o f g e n J " " ^ ^ " ^ - ^ - - • ( - 0 ,
° ' ""^'elle .ylostells .as „ , , " " " ' ' ° " " ' ' ' ^ ^ " - - " ^ '™e " 35 prolonged to ?«on
- P - V e M . e . , h e , a h o r a , o ^ _ ^ ' - ^ ^ ^ - ^^-^^ ^ays
- 'oe . 0 0 . , a,so reporfe. . a f l ' " " " " ^ ^ ^ ^
- • • - " . c . . . . . _ ^ ^ ^^^^^^^^^^^^
° - - ^^e .eve,op.en,a, . .af ion f l " ^ ' ^ " '^^ ^ - - d ----o.aofapp„oa::: ^^^^^^ a/., 2003). ^^°'°"9-^ significanfiy (Wang e,
-88-
th
Oiscussum
Spinosad
The topical application of different sublethal concentration of
spinosad viz., 0.1%, 0.08%, 0.06%, 0.04% and 0.02% at two different
temperature/photoperiod and constant relative humidity (60-70%) on 4
instar nymphs of Dysdercus koenigii causes instant mortality as well as
mortality upto adult emergence. The initial mortalities within 24 hour and
upto adult emergence increased with increasing sublethal
concentrations. The initial mortality within 24 hour and the mortality
upto adult emergence was comparatively higher at 30±1°C/14L: 10D as
compared to 25±1°C/12L: 12D temperature and photoperiod. The
topical application of the highest selected sublethal concentration i.e.
0.1% of spinosad at 30±1°C/14L:10D temperature caused 27.76% initial
nymphal mortality within 24 hour, however, at 25±1°C/12L:12D the initial
nymphal loss within 24 hrs was 13.33% less as compared to higher
temperature/ photoperiod. The topical application of the lowest selected
sublethal concentration (i.e. 0.02%) of spinosad at 30±1°C/14L: 10D
produced 2.2% initial nymphal loss, whereas, at 25±1°C/12L. 12D
temperature/ photoperiod it was only 1.1%. However, the total nymphal
death uptp adult emergence following the topical application of 0.1% of
spinosad at 30±1°C/14L: 10D and 25±1°C/12L: 12D
temperature/photoperiod goes to 56.67 and 31.10% respectively as
compared to control, whereas, topical application of the lowest selected
-89-
(Discussion
sublethal concentration (0.02%) at the same condition produced only
5.53 and -2.20% loss respectively. Athanassiou et. al,. (2008) studied
the influence of temperature and humidity on the efficacy of spinosad
against four stored-grain beetle species viz., lesser grain borer,
Rhyzopertha dominica (F.), rice weevil, Sitophilus oryzae (L.), confused
flour beetle, Tribolium confusum Jacquelin du Val and Prostephanus
truncates (Horn) and concluded that there was a significant impact of
temperature and relative humidity on spinosad efficacy against all
species tested. The influence of these two parameters is complex since
it varied according to the dose rate and the exposure interval. Kowalska
(2008) studied the activity of three different concentrations of spinosad
viz., 0.2, 0.1 and 0.05% against the larvae and adults of colerado potato
beetle Leptinotarsa decemlineata at three different temperature viz., 15,
20 and 25°C in laboratory and concluded that temperature play an
important role on the efficacy of spinosad and all the concentrations
tested caused mortality, which increased as concentration of spinosad
increased. Fang and Subramanyam (2003) found that 0.1 ppm of
spinosad on wheat caused 100% mortality of R. dominica adults and
notably suppressed progeny production. Nayak et. al., 2005 also tested
several R. dominica strains with various susceptibility levels to some of
the most commonly used grain protectant and found that 0.1 ppm of
spinosad provided 93-99% adult mortality after 7 days of exposure,
-90-
(Discussum
regardless of the resistance strain to other pesticide of the beetle
tested. Irani and Asghar, 2007 used a commercial formulation of
spinosad, Tracer® against 7-14 days old adults of rice weevil Sitophilus
oiyzae and red flour beetle Tribolium castaneum at three different
temperature (i.e. 24, 28 and 32°C and 65±5% RH) and observed that
there was a dose-dependent response and mortality increased, with
increasing the dosages in all temperatures in both species. Eger et al.,
(1998) also reported the same results in which mortality of Frankliniella
sp. showed a concentration dependent response. Likewise, Toews and
Subramanyam (2003) also recorded the same trend and reported that
the mortality of T. Castaneum increased with increasing dosage of
insecticide. A biochemical reason for this similarity is imposing of the
same enzymes with spinosad in these experiments.
The exposure of spinosad to the 4* instar nymphs of O. Koenigii
resulted in the emergence of malformed adults (Plate-3 & 4). This
percentage of malformation increased with increasing sublethal
concentration at both 30±1°C/14L: 10D and 25±1°C/12L: 12D
temperature/photoperiod with constant relative humidity. The
percentage of malformation at the highest selected sublethal
concentration was 17.76 and 21.1% with respect to 30±1°C/14L: 10D
and 25±1°C/12L: 12D temperature/photoperiod, whereas, at the lowest
selected dose it was only 20% and 15.53% respectively. El-Barkey et.
-91-
Q)iscusswn
al., (2009) studied the biological effects of Radiant and Hexaflumuron
against the eggs of Pectinophora gossypiella (Saunders) and reported
that both compound caused malformation in larvae, pupae and adults
emerged from treated eggs. Sublethal effects of insecticide on the
malformation and reproductive capacity reduction observed in the
survivors could have a negative impact on insect population dynamics
(Trisyono and Chippendale, 1998; Lopez and Latheef, 1999; Gobbi et.
al., 2000 Knight, 2000; Pineda et. al., 2004).
The average egg production as well as average egg hatching of
D. Koenigii were also adversely affected following the topical application
of sublethal concentrations of spinosad at both 30±1°C/14L:10D and
25±1°C/12L:12D temperature/photoperiod. The loss in fecundity and
fertility was also concentration based and increased with increasing
concentration. The reduction in average egg production of the females
following the topical application of the highest selected sublethal
concentration at two above said temperatures/photoperiods was 54.24
and 61.68%, whereas, at the lowest sublethal concentration the loss in
fecundity was 28.77 and 19.75% respectively. Similarly, a significant
reduction of fecundity was observed when Ceratitis capitata (Diptera;
tephritidae) adults ingested O.lmg/liter spinosad (Adan et. al., 1996).
Temerak, 2007 also found that Radiant at 5.76 gram active/ HA showed
100% mortality of entire hatched egg masses of Spodoptera littorilis
-92-
Q>iscussitm
after the spray in the field. The fertility of the female which emerged
from the 4 ^ instar nymphs treated with 0.1% of spinosad at both
30±1°C/14L:10D and 25±1°C/12L:12D temperature/photoperiod was
reduced by 65.47 and 72.12%, whereas, at lowest concentration, the
drop in fertility was 32.14 and 23.81% respectively as compared to
control. Similarly Yin et. al., (2008) recorded that the adult longevity,
population growth and fecundity of Plutella xylostella was strongly
reduced when the larvae were treated with spinosad. Amer (2004)
recorded that spinosad caused reduction in adult longevity, fecundity
and fertility of P. gossypiella. Also, Liu and Trumble (2005) and
Zaiiznaik and Nugegod (2006) reported that spinosad high affected on
fertility and fecundity of Bactehcerca cockerelli. Egg fertility was
reduced in Helicoverpa zea (Boddie) when females were fed with
concentrations <1 mg (Al)/liter and then paired with untreated males
(Lopez and Latheef 1999).
Besides the acute toxicity produced by the present chemicals, the
biology and physiology of the survived insects were also adversely
affected by their sublethal dosages. Some earlier reports suggest that
the treatment with chemicals may enhance the survival duration of the
affected insects (Knutson, 1955; Katiyar & Lemonde, 1972; Abo-Elghar
et. al., 1972; Parker et. al., 1976; Khowaja et. al., 1991, '92; '93; '94;
2001, '02, '05), whereas, other workers have the opinion that treatment
-93-
Qiscussion
with chemicals reduced the longevity of the survived insects (Haynes,
1988; Mackenzie & Winston, 1989; Abd-Elghafar and Appel, 1992). The
present observations on D. koenigii also prove that the topical
application of sublethal concentrations of spinosad at both 30±1°C714L:
10D and 25±1°C/12L: 12D temperature/photoperiod enhance the
longevity of all the stages of survived treated insects. The topical
application of the highest selected sublethal concentration i.e. 0.1%
spinosad on 4* instar nymphs at both 30±1°C/14L: 10D and
25±1°C/12L: 12D temperature/photoperiod showed average
enhancement of nymphal duration by 38.09 and 30.76% respectively as
compared to their control, while, at the lowest concentration (i.e. 0.02%)
the prolongation of nymphal duration was 4.76 and 3.84% respectively.
El-Barkey et. al., (2009) revealed that the exposure of Pectinophora
gossypiella to radiant and hexaflumur significantly prolonged the
duration of the larval stage than that of untreated check. The longevity
of the 5* instar nymphs which emerged from the 4* instar treated
survived nymphs following the application of 0.1% spinosad was also
enhanced by 40.74 and 42.30% respectively as compared to control,
while at the lowest concentration, the survival duration was increased
by 7.40 and 11.53% respectively as compared with control. Yin et. al.,
(2008) reported that prolonged in the immature stage and the survival
rate of Plutella xylostella was lower in the LC25 and LC50 for spinosad.
-94-
"•'""'—* ..«„«:!:, 7""-»«~ wnicn emerged frnm •»,
'"oreasedby 4.47 and , 420/ re ' " ' " ° ' '^"^^'^^ *-=
•^^' ^P-sad a, s.5,e,.a, o o n c e n r r ' ' ' " ' " ' ' ' ^ ' ° ° ' ' ^ ^ ° - ' '
°^ - • - ,a.ae a . . . . ^ ^ " " ^ " ^ ^ - ' - ^ « -
' - - ' - ° - . . a . a M . e , o p . e n , o " " ' ° " " " ^ • ^ - -
- ^ - - - . a s c o n e e . a . . . p . ^ ^ ^ " ^ ^ " ^ " ' ^ ^ ^ • ^ ^
<=y» ion (malathion) was 33 90/. " ' " " " ' " * ° ""^-^^
•• 1991). Acord,ng ,0 Conney et a/ nsee^ •
'"^*«ed hy*oxy,atlon process of so.e , ' ' " ----0---.one, j : ; : r ' ° '--"-- - K a « y a r a n d . e ™ n d e ( , 9 7 2 , o " ' * " ^ " ' ^ " ' " " -
(1972) oPse^ed .onoc.o.ophos, ga.dona
-95-
Q>iscussum
and aldicarb to act as antiacetylchoiinestrase agents resulting in greater
prolongation of the larval period of T. confusum. Perhaps, the increase
in longevity of D. koenigii may be attributed to the
antiacetylchoiinestrase activity when spinosad was applied on the
nymphs.
The present finding of these biological parameters can be helpful
in manipulating some of the chemical methods to keep the pest
population at the minimum. The recommended doses should be
regarded temperature/photoperiod specific and some consideration
must be given to decline in effectiveness with lowehng of temperature. It
may well not prove feasible to compensate for this decline by increasing
the dose applied since this could put spray operator at greater risk, lead
to residue hazards or prove uneconomic. Certainly the less effective
insecticides should not be used at all where temperatures are very low.
In some situation, it may be possible to raise the temperature of the
treated area sufficiently to activate the insects so that they may pick up
a lethal dose.
-96-
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