II.

REVIEW OF LITERATURE

The literature reviewed here pertains to the production of cotton, Bt cotton, and

different types of insecticides such as Neonicotinoids viz., imidacloprid, acetamiprid,

thiacloprid, thiamethoxam, clothianidin, used to control the sucking insect pests of Bt

cotton, laboratory bioassay, field experiments on cotton sucking pests, cotton aphids,

jassids, whiteflies, the safety of neonicotinoids to natural enemies, phytotoxicity of

neonicotinoids and harvest time residues of promising neonicotinoids.

Production of cotton

The spectacular achievement in the production and productivity of cotton in India

owes itself to the introduction of Bt cotton in 2002. Cotton production suffered huge losses

due to its susceptibility to insect pests, particularly bollworms, before the introduction of

Bt cotton in India. The cotton bollworm complex comprises the American bollworm,

7
Helicoverpa armigera; pink bollworm, Pectinophora gossypiella; spiny bollworm, Earias

insulana and spotted bollworm, Earias vittella were reported to cause monetary losses of

at least US$ 300 million/annum.

The cost of 21,500 metric tonnes of insecticides (active ingredient) used on cotton

in India in 2001 was US$340 million, accounting for about half of the total value of

insecticides used on all crops in India (US$660). Further, the most destructive cotton pest,

Helicoverpa armigera, is known to have developed resistance against most of the

recommended insecticides (Ramasubramanian, 2004) and forced farmers to apply as many

as 10-16 sprays. At this juncture, the Indian government approved for commercial

cultivation of Bt cotton in 2002 to three cotton hybrids, MECH-12 Bt, MECH-162 Bt and

MECH-184 Bt. The insect resistance in these hybrids was introgressed from Bt gene

cry1AC containing Cocker-312 (event MON 531, Bollgard I) developed by Monsanto, the

USA, into parental lines of Mahyco’s proprietary hybrids (Karihaloo and Kumar, 2009).

Sucking pests on Bt cotton:

Continuous cultivation of Bt cotton since its inception in 2002 has led to an

increased incidence of sucking pests and other pests of minor importance (Nagrare et al.,

2009) as the Cry1Ac toxin expressed by Bt cotton is not toxic to sucking pests. Since the

sucking pests have enormous potential of becoming major pests of cotton, the identification

of selective and effective insecticides for managing these pests on Bt cotton deserves utmost

importance.

The economically important sucking insect pests of cotton are aphid, Aphis gossypii

Glover; jassid, Amrasca devastans (Distant); thrips, Thrips tabaci Lindeman; and whitefly,

Bemisia tabaci Gennadius.

Cotton is affected by numerous insect pests and causes about 15-40% yield loss in

India through heavy infestation and the loss was due to leafhoppers and bollworms complex

8
(Sohi, 1964). Regupathy (1981) reported that the growth and dry matter production of the

cotton plant was reduced to the extent of 38 to 44% by jassid and aphid incidence.

Bandhanial et al. (1993) reported that avoidable losses in cotton due to sucking

pests’ infestation were 13.55. Sarnaik et al. (1994) reported that the avoidable yield loss

in cotton variety PKV Hy-2 was 49.65% or 5.64 q/ha loss in yield due to sucking pests and

bollworms together. Dhaliwal and Arora (1996) also reported a yield loss of 50% due to

insect pests on cotton.

Due to the continuous use of conventional insecticides, sucking insect pests like B.

tabaci, A. devastans and A. gossypii have developed varying levels of resistance to almost

all the conventional insecticides like organophosphates, carbamates, synthetic pyrethroids

(Ahmad et al., 2000, 2001 and 2003). This led to several problems like toxic residues,

elimination of natural enemies, environmental disharmony and development of resistance.

To overcome those problems, identifying new molecules with selective insecticidal

properties, low toxicity to non-target, well-suited IPM practices, and an effective chemical

management strategy against sucking pests utilizing selected neonicotinoids with low rates

and novel mode of action attracted the attention. Its broad-spectrum activity against sucking

pests and systemic properties appear to be helpful for the management of A. gossypii, A.

devastans and B. tabaci.

2.1. NEONICOTINOIDS

Neonicotinoids are a class of synthetic insecticides widely used in agriculture to

protect a range of crops from insect attacks and are particularly effective in controlling

sucking insects like aphids, whiteflies and plant hoppers and chewing insects like beetles

and the larvae of some moths. Apart from its use in agriculture, human and animal parasites

like bed bugs and fleas are also controlled with the help of neonicotinoids. Neonicotinoids

are chemically related to nicotine, a potent natural insecticide produced by tobacco plants

9
as a defense against herbivorous insects. The first neonicotinoid (imidacloprid) was

introduced in 1991 and there are now eight commercial compounds (Bass and Field, 2018).

The first neonicotinoid, imidacloprid, was introduced by Bayer CropScience. While

Syngenta introduced thiamethoxam. Sumitomo Chemical and Bayer CropScience have

introduced clothianidin and acetamiprid by Nippon Soda, thiacloprid, dinotefuran and

nitenpyram were introduced by Bayer CropScience, Mitsui chemicals and Sumitomo

chemicals, respectively (Jechke et al., 2011).

Dow Agrosciences India Pvt. Ltd introduced the eighth neonicotinoid, Sulfoxaflor,

in India in 2017. In August 2018, Shanghai Shengnong Pesticide was granted full

registration of cycloxaprid technical and cycloxaprid 25% WP for application on rice to

control planthoppers and cabbage aphids and whiteflies in China.

Neonicotinoids, the newer major class of insecticides, have outstanding potency and

systemic action for crop protection against piercing-sucking pests. Some popular

neonicotinoids are acetamiprid, clothianidin, imidacloprid, thiacloprid, and thiamethoxam.

They possess lower mammalian toxicity, fewer resurgence problems, environmental

protection, pest management selectivity and less toxicity to natural enemies (Kunkel et al.,

1999).

The neonicotinoids that mimic nicotine are known for their intense activity against

sucking pests (Bethke and Redak, 1997; Horowitz et al., 1998; Senn et al., 1998). Even

though the neonicotinoids do not kill the sucking insects immediately, they cause rapid

cessation of feeding (Harrewijn et al., 1998).

Neonicotinoid selectivity results from a higher affinity to the insect nicotinic

acetylcholine receptor (nAChR) compared to vertebrates, in contrast to the original

nicotinic compound (Ishaaya et al., 2007). Neonicotinoid insecticides' extraordinary

spectrum of efficacy, together with full exploitation of the nAChR, effective systemic

10
action in plants, long-lasting effect and versatile uses and application methods have

contributed to the unique success of this class of insecticide (Elbert et al., 2008)

Neonicotinoids are broad-spectrum, systemic compounds that exhibit activity

against sucking insects. These are readily absorbed by plants and act quickly, at low doses

as compared to other insecticides (Sympathy and Rai, 2006). The expanding importance of

crops expressing Bt δ-endotoxin encourages neonicotinoid use because the types of pests

not controlled by the endotoxin are often susceptible to neonicotinoids (Kagabu, 2005 and

2003). Neonicotinoids are used at a lower dosage as compared to conventional insecticides.

Neonicotinoids are structurally distinct from all other synthetic and botanical

pesticides and exhibit favourable selectivity. Neonicotinoids are increasingly replacing

organophosphates and methyl carbamates to control piercing-sucking insect pests.

Neonicotinoids have a unique mode of action as compared to organophosphates,

carbamates and synthetic pyrethroids. Since they bind at a specific site, the post-synaptic

nAChR and hence no records of cross-resistance to the carbamates, synthetic pyrethroids

and organophosphates, thus making them essential for the management of insecticide

resistance. These are highly toxic to insects compared to conventional insecticides, and they

provide lower toxicity to mammals (Ajaz et al., 2018). Like the naturally occurring alkaloid

(S)-(−)-nicotine, all neonicotinoids act selectively on the insect central nervous system

(CNS) as agonists of the post-synaptic nicotinic acetylcholine receptors (nAChRs), 2–5

their molecular target site (Bai et al., 1991).

Like nicotine, neonicotinoids act on the insect nervous system, targeting the

nicotinic acetylcholine receptor (nAChR), an ion channel that plays a crucial role in nerve

signalling. All neonicotinoids are highly specific for insect nAChRs and have low toxicity

to vertebrates. Neonicotinoids are water-soluble and can be taken up by plants and

distributed to all tissues. The systemic properties of neonicotinoids ensure that they can be

11
applied to the seed before planting. The seed coating will protect the crop during the initial

growth stages and minimize foliar spray applications. Even though many insect species are

still successfully controlled by neonicotinoids, their widespread and extensive use has

resulted in the evolution of resistance in some economically important insect pests,

including whiteflies, aphids, planthoppers and beetles (Bass and Field, 2018).

The safety and effectiveness of neonicotinoid insecticides were attributed to a high

selectivity for insects compared to non-target vertebrate nAChRs, and its fast penetration

into the insect central nervous system, but most important is the difference in target site

interactions (Methfessel, 1992; Tomizawa and Casida, 2001; Matsuda et al., 2001).

Since their introduction in the 1990s, neonicotinoids have become the most widely

used insecticide in the world, accounting for over 25% of the global insecticide market and

approximately $4 billion in 2014 (Bass et al., 2015). Neonicotinoid insecticides are highly

selective agonists of insect nicotinic acetylcholine receptors and provide farmers with

invaluable, dependable and highly effective tools against some of the world's most

destructive and economically important crop pests.

2.1.1. IMIDACLOPRID

Imidacloprid with Chemical Abstract Name, 1-[(6-Chloro-3-pyridinyl)methyl]-N-

nitro-2-imidazolidinimine) was a highly effective systemic insecticide developed in the

U.S. by Miles Inc. and by Bayer AG worldwide. Imidacloprid is a systemic and contact

insecticide exhibiting low mammalian toxicity, with primary activity on sucking insects

such as aphids, leafhoppers and planthoppers, thrips and whiteflies including strains

resistant to conventional chemistries. With excellent systemic and good residual

characteristics, imidacloprid is especially appropriate for seed treatment and soil

application (Elbert et al., 1998). Effective early-season control with long-lasting protection

is achieved in crops such as cereals, corn, cotton, potatoes, rice, sorghum and many

12
vegetables. Pests attacking later in the season can be controlled by foliar applications in the

crops mentioned above, as well as in citrus, deciduous fruits, grapes and other crops

(Mullins, 1993). It has also been shown that it kills adult insects at comparatively high

doses and has ovicidal effects. Imidacloprid is a nicotinic acetylcholine receptor stimulator.

Its mechanism of action differs from that of conventional insecticides. It gives excellent

control of all resistant populations investigated hitherto (Elbert, 1991).

2.1.2. ACETAMIPRID

Acetamiprid with the chemical abstract name, (E)-N1-[(6-chloro-3-pyridyl)methyI]-

N 2-cyano-N 1-methylactamidile, is a novel insecticide developed by Nippon Soda Co., Ltd.

Although the compound belongs to the neonicotinoids, it possesses characteristic

insecticidal properties different from others in the same category of chemical structure.

Acetamiprid has a cyanoamidine structure, which contains a 6-chloro-3-pyridylmethyl

moiety.

Acetamiprid shows excellent activities against Hemiptera and Thysanoptera, as do

other neonicotinoids; it exhibits excellent activity against Lepidoptera, and the insecticide

is applicable for controlling pests of vegetables, fruit trees, the tea tree, and so on. (Yamada

et al., 1999).

2.1.3. THIACLOPRID

Thiacloprid with the chemical abstract name, [3-[(6-Chloro-3-pyridinyl)methyl]-2-

thiazolidinylidene]cyanamide is an insecticide of the neonicotinoid class. Its mechanism of

action is similar to other neonicotinoids and involves disruption of the insect's nervous

system by stimulating nicotinic acetylcholine receptors. Thiacloprid was developed by

Bayer CropScience for use on crops to control a variety of sucking and chewing insects,

primarily aphids and whiteflies. Due to its rain-fastness property, thiacloprid is stable even

under conditions of heavy rains and sunlight, providing longer persistence.

13
 Thiacloprid is an antagonist to the nicotinic acetylcholine receptor in the central

nervous system. It disturbs the proper signal transmission system leading to the excitation

of the nerve cell. Consequently, a disorder of the nervous system occurs, finally leading to

the treated insect's death. Thiacloprid is effective against insect pests like aphids, jassids,

thrips, whiteflies, some coleopterans, hemipterans and lepidopteran caterpillars (Schuld

and Schmuck, 2000).

2.1.4. THIAMETHOXAM

Thiamethoxam with Chemical Abstract Name, 3-((2-chloro-5-thiazolyl)methyl)

tetrahydro-5-methyl-N-nitro-4H-1,3,5-oxadiazin-4-imine, was the first commercially

available second-generation neonicotinoid and belonged to the thianicotinyl sub-class. The

compound has broad-spectrum insecticidal activity and offers excellent control of a wide

variety of commercially important pests in many crops (Maienfisch et al., 2001)

Thiamethoxam acts by binding to nicotinic acetylcholine receptors. It exhibits

exceptional systemic characteristics and controls a broad range of commercially important

pests, such as aphids, jassids, whiteflies, thrips, rice hoppers, Colorado potato beetle and

wireworms, as well as some lepidopteran species. In addition, a strong preventative effect

on some virus transmissions has been demonstrated. Thiamethoxam is developed both for

foliar/soil applications and as a seed treatment for use in most crops all over the world. Low

use rates, flexible application methods, excellent efficacy, long-lasting residual activity and

favourable safety profile make this new insecticide well-suited for modern integrated pest

management programs in many cropping systems (Maienfisch et al., 2001)

2.1.5. CLOTHIANIDIN

Clothianidin with chemical abstract name, 1-(2-Chloro-1,3-thiazol-5-methyl)-3-

methyl-2-nitroguanidine, a neonicotinoid insecticide, has been found by former Agro

Division, Takeda Chemical Industries, Ltd. (Sumitomo Chemical Co., Ltd., at present) and

14
co-developed with Bayer CropScience. During the studies on neonicotinoid insecticides,

nitenpyram (an open-chain nitromethylene derivative) was prepared first, showing potent

activity against Hemiptera and Thysanoptera pests and its modification led to clothianidin

(a nitroguanidine derivative).

Clothianidin exhibits excellent control efficacies in small amounts for a wide

variety of insect pests such as Hemiptera, Thysanoptera, Coleoptera, Lepidoptera, and

Diptera for the long term, with excellent systemic action and by a variety of application

methods (Hideki, 2011)

2. 2. LAB BIOASSAY

Tufail et al. (1995) reported that imidacloprid was very effective in the control

of sucking insect pests, especially cotton jassid. Naimatullah et al. (1998) reported that

different insecticides, including imidacloprid showed a considerable effect in suppressing

the sucking insect pest, e.g., Amrasca devastans, Aphis gossypii and Bemisia tabaci. Saleem

and Khan (2001) described that imidacloprid was very effective for controlling cotton

jassids.

Misra (2002) reported that imidacloprid and thiamethoxam showed significantly

higher performance in controlling jassids and aphids. Imidacloprid was highly effective

against cotton jassids (Amrasca devastans Ishida) after applying insecticides at different

time intervals, e.g., at 72, 96 and 240 hrs (Razaq et al., 2003). Imidacloprid application was

highly effective against cotton whiteflies (Bemisia tabaci) and jassids (Amrasca bigutulla

bigutulla) by providing control for up to 168 hrs (Asif et al., 2008).

El-Dewy (2006) reported that imidacloprid was found to be a very effective

insecticide against jassids and whitefly. A higher insecticide concentration was more

effective than lower and standard concentrations for controlling cotton jassids (Amrasca

bigutulla bigutulla). With the application of acetamiprid at 0.02%, effective control of

15
mustard aphid was achieved (Singh and Verma, 2008; Singh and Singh, 2009; Dhaka et al.,

2009; Mandal and Mandal, 2010).

Shreevani et al. (2012) reported that the cumulative toxic effect at 48 hrs of spray

with highest for thiamethoxam 25 WDG (82.67%), followed by imidacloprid 17.8 SL and

clothianidin 50 WDG. The standard check treatments viz., oxydemeton methyl 25 EC and

dimethoate 30 EC recorded lower percent mortality. The toxic effects were persistent even

at 72 and 96 hrs after application. Thiamethoxam 25 WDG could achieve 100% mortality

of jassids at 72 hrs, while imidacloprid 17.8 SL and clothianidin 50 WDG took 96 hrs to

achieve a similar level of mortality.

Shreevani et al. (2014) reported that the percent mortality of leafhoppers was found

to be maximum at 50.67% with thiamethoxam followed by imidacloprid (46.67%) and

clothianidin (37.33%) at 24 hrs after treatment. At 48 hrs of treatment highest percent

mortality was noticed in thiamethoxam spray treatment (82.67%) followed by imidacloprid

and clothianidin. The standard check treatments, viz., oxydemeton methyl and dimethoate

could achieve low percent mortality of only 21.33% and 16.00%, respectively. Percent

mortality of leafhoppers recorded at 72 and 96 hrs after the application followed the same

trend as that of 48 hrs. Thiamethoxam treatment achieved 100% mortality of leafhoppers

at 72 hrs while imidacloprid and clothianidin took 96 hrs to achieve 100% mortality. The

standard check treatments viz., oxydemeton methyl, dimethoate and pyriproxyfen could not

achieve 100% mortality even after 96 hrs of application.

2.3. FIELD EXPERIMENTS

2.3.1. Cotton Sucking pests

Reddy et al. (1986) reported a yield loss of 10 – 25% during 1984-85 and 20 – 45%

during 1985-86 in seed cotton due to whitefly in Andhra Pradesh. Dhawan et al. (1988)

reported an avoidable loss of 1.10 q/ha due to jassid attaching to seed cotton. Sarath Babu

16
and Gupta (1988) reported that the soil application of systemic granular insecticides or

spraying of insecticide preparations remains the traditional approach to controlling sucking

pests. However, they adversely affect the population of non-target soil organisms.

Attique and Ghaffer (1996) carried out a cotton field trial with imidacloprid and

reported that the attack of Thrips tabaci Lindeman was reduced by up to four weeks. Seed

treatment and foliar application of imidacloprid significantly decreased the sucking pests’

population in cotton, stated Kumar (1998). Sucking pests’ infestation during the early

stages of the cotton crop resulted in more rounds of insecticides than required to protect the

crop, causing ecological imbalances and many other associated problems (Gupta et al.,

1998).

As per International Crop Advisory Committee (ICAC) (1998) report, about 50%

of the cotton yields in the world are attributed to using of agrochemicals. Cotton sucking

insect pests viz., jassid, A. devastans, aphid, A. gossypii, whitefly, B. tabaci and thrips, T.

tabaci can pose a serious threat and cause severe damage to the crop at the early stage of

the cotton crop and can also affect the crop stand and yield of cotton. Patil (1998) reported

a reduction in the cotton crop yield to 21.2% due to heavy infestation.

Kumar et al. (2009) reported that spirotetromat at 75 g a.i./ha and imidacloprid at

25 g a.i./ha were equally effective in managing cotton whitefly.

Rathod (2003) reported that seed treatment with imidacloprid 70 WS at doses of 5,

7.5 and 10 g per kg of seeds significantly reduced the population of cotton-sucking pests

like aphids, jassids and thrips when compared to control while increasing the yield of seed

cotton. Acetamiprid and thiamethoxam were the most promising insecticides against

whitefly (Muhammad et al., 2004).

Aslam et al. (2004) evaluated the efficacy of seven insecticides against cotton

sucking pests and reported that imidacloprid was the most effective insecticide for jassids

17
for up to seven days. During its early growth stage, the protection of the cotton crop from

the sucking pests is crucial and vital because it is a fact that a good plant stands at the initial

stage results in good produce (Rajeshwaran et al., 2005). Raghuraman and Gupta (2005)

studied the efficacy of neonicotinoids against whitefly, Bemisia tabaci Gennadius in cotton

and indicated that acetamiprid and imidacloprid were good substitutes for conventional

insecticides. Acetamiprid at 40 g a.i./ha and imidacloprid at 100 g a.i./ha was found to be

the most effective treatments against whitefly on cotton.

Patil et al. (2007) reported that results of pooled analysis data of two sprays of

clothianidin 50% WDG at 20 and 25 g a.i./ha rendered excellent crop protection against the

early season sucking pests infesting cotton hybrid DHH-11. Aggarwal et al. (2007) reported

that the population of sucking pests was significantly higher in Bt cotton hybrids compared

to non-Bt hybrids.

Jeyakumar et al. (2008) reported that even though Bt cotton was successful in the

management of chewing insect pests like bollworms, it has attracted other insect pests,

especially sucking pests due to a reduction in pesticide sprays during the early stage of the

crop. Conventional organophosphates and carbamates have shown resistance to whitefly

and leafhopper in cotton (Ahmad et al., 2010). Chavan et al. (2010) reported that 28.13%

of yield loss could be avoided due to major sucking pests in cotton.

Shera (2012) reported that the extent of avoidable losses was 252 kg/ha (11.9 %) in

MRC 6301 Bt as against 263 kg/ha (11.2%) in RCH 134 Bt and 290 kg/ha (16.2 %) in NCS

145 BG II. Bajya et al. (2013) reported that thiamethoxam75% SG at 94 and 113, g a.i. per

ha were found to be more efficient in reducing jassids, whiteflies and thrips population

compared to imidaclorpid 17.8% SL and thiamethoxam 25% WG over untreated control.

El-Naggar and Zidan (2013) reported that treatments with imidacloprid and thiamethoxam

as foliar applications were highly effective against aphids, for up to 14 days in the case of

18
jassids, while the effect was moderate on the whitefly population (mature and immature

stages). Imidacloprid had more initial and residual effects than thiamethoxam against

jassids.

Bharpoda et al. (2014) reported that among the different insecticides, imidacloprid

17.8 SL at 0.008% (7.50 aphid and 1.47 whitefly/leaf), thiamethoxam 25 WG at 0.0125%

(1.22 leafhopper/leaf) and diafenthiuron 50 WP at 0.05% (1.43 thrips/ leaf) found more

effective and safer to the natural enemies viz., Chrysoperla carnea (adult), spiders and

coccinellids (grubs and adult). The highest seed cotton yield (30.81 q/ha) was harvested

from crops treated with imidacloprid followed by clothianidin (27.34 q/ha), diafenthiuron

(26.93 q/ha), thiamethoxam (26.01 q/ha) and acetamiprid (25.68 q/ha).

Kadam et al. (2014) reported significantly lowest population of sucking pests per

three leaves was recorded in nitenpyram 10 per cent WSG at 100 g a.i./ha, dinotefuran 20

per cent SG at 50 g a.i./ha and clothianidin 50 per cent WDG at 20 g a.i./ha were the most

effective treatments in reducing the incidence of sucking pests on Bt cotton as compared to

acetamiprid 20 per cent SP at 20 g a.i./ha, imidacloprid 17.8 SL, thiamethoxam 25 per cent

WS at 25 g a.i./ha and thiacloprid 21.7 per cent SC at 30 g a.i./ha.

Deepika et al. (2015) reported that Bt genotypes recorded a higher whitefly

population than non-Bt genotypes. Asif et al. (2016) reported that imidacloprid followed

by nitenpyram proved to be most effective for significantly reducing whitefly and thrips

populations. Nitenpyram had the highest percentage reduction (73.80%) against jassid on

the 7th day after application, but that was non-significantly different from imidacloprid

(63.49%).

Malik et al. (2018) reported that the application of acetamiprid 20 SP at 125 ml per

acre against thrips after one week of application followed by nitenpyram at 200 ml and

profenophos at 800 ml per acre gave the best results.

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2.3.1.1. Cotton Aphid, Aphis gossypii Glover (Hemiptera: Aphididae)

Among the cotton pests, the cotton aphid, A. gossypii is an important pest that

infests cotton crops during the early crop season, delaying normal plant development.

Without human intervention, the aphid infestation might extend through the development

of the plants. Consequent to the infestation with cotton aphids, the cotton crop exhibited

reduced development and curled leaves, notably the young leaves that are essential for the

growth of the main stem and the leaves of the reproductive branches of the cotton crops

(Ebert, 2008; Leclant and Deguine, 1994).

Apart from directly damaging the plant due to feeding, aphids also secrete

honeydew and drops on the leaves and the open lint, seriously affecting the cotton yield.

The honeydew favours the development of black sooty mould fungus, which affects plant

development through inhibition of photosynthesis and results in sticky lint, which affects

the spinning process at the textile mills (Deguine et al., 2000). Within a short span, cotton

aphids can develop into large colonies due to the intrinsic biotic characteristics of the

aphids, such as rapid development and parthenogenic mode of reproduction in the tropical

regions, which are enhanced due to high temperatures and water stress (Godfrey et al.,

2000; Vanemden and Harrington, 2007).

Resistance to organophosphates in aphids, A. gossypii was first reported in 1961

(Kung et al., 1961). Aphids were reported to be resistant to conventional chemical

insecticides like endosulfan, cypermethrin, deltamethrin and fenvalerate (Delorme et al.,

1997; Villate et al., 1999; Ahmed et al., 1999; Herron et al., 2001), monocrotophos and

dimethoate (Deguine, 1996; Nibouche et al., 2002) and carbamates (Furk et al., 1980;

Bobert et al., 1994).

Williams (2001) reported that in the 2000 cultivation season, cotton aphids infested

approximately 78% of the cotton acreage in the United States, resulting in an estimated loss

20
of 130,000 bales. Cotton aphids injure cotton by continuously feeding on fluids in plant

phloem systems (Rohfritsch, 1990). The phloem-feeding can result in lesser boll formation,

boll retention and reduced cotton boll and lint weight (Bagwell et al., 1991; Fuchs and

Minzenmayer, 1995).

In the United States of America, cotton yield losses associated with cotton aphids

have been documented in several states (Andrews and Kitten, 1989; Layton et al., 1996).

Bagwell et al. (1991) reported that cotton plants heavily infested with cotton aphids were

shorter and had fewer main stem nodes and bolls than plants with low densities of aphids.

Boll retention at first and second fruiting positions on the lowest five sympodial branches

was reduced. Rosenheim et al. (1997) reported that cotton aphid infestations on pre-

flowering stage plants impacted the plant architecture compared to non-infested plants.

Cotton aphids’ infestation has reduced leaf area by 58% and shoot biomass by 45%.

Cotton plants infested with cotton aphids were stunted with few vegetative branches than

non-infested cotton plants. High densities of aphid infestation can commonly occur as

resurgent populations consequent to the applications of selected insecticides for other insect

pests (Slosser et al., 1989, Torrey et al., 2000).

Ghosal et al. (2013) reported that imidacloprid 17.8 SL at 50 g a.i./ha was found to

be the most effective neonicotinoid insecticide against aphids. It recorded the least aphid

infestation and an 84.54% reduction of the population over control. The other two

neonicotinoids, thiamethoxam 25WG at 50g a.i./ha and acetamiprid 20SP at 40 g a.i./ha

were also found at par with imidacloprid and showed better results than acephate 75WP

and dimethoate 30EC.

21
2.3.1.2. Jassid, Amrasca devastans (Dist.) (Hemiptera: Cicadellidae)

Except during the winter season, jassid breeds throughout the year. In spring, cotton

jassid migrates to okra and starts breeding. The pest completes seven generations in a year.

The yield losses due to jassids have been reported to be 1.19 q per ha (Dhawan et al., 1988).

About 25-45% loss of cotton yield was reported due to the attack of the leafhopper

(Bhat et al. 1986). Dhawan and Sidhu (1986) reported that the cotton leafhopper, A.

biguttula biguttula (Ishida) is a serious sucking insect pest as it causes a loss of 9% seed

cotton yield.

The yield loss due to leafhopper attack was 108kg/ha in cotton cultivar F414, a

highly tolerant variety and 160kg/ha in J 205 cotton variety, which was highly susceptible

to leafhoppers. Severe attacks by leafhoppers on cotton crops resulted in a 35%

(Neelakantan, 1957) and an 18.8% (Javed et al., 1992) reduction in yields.

Leafhopper, A. devastans was reported to have developed resistance to conventional

insecticides like endosulfan, monocrotophos, cypermethrin, phosphamidon, dimethoate,

methyl demeton and acephate (Santhini and Uthamasamy, 1997; Chalam and Subbaratnam,

1999; Jeyapradeepa, 2000; Chalam et al., 2001 and Praveen, 2003).

Bt cotton has been commercially cultivated in India since 2002 with the

expectations of a reduction in the number of pesticide applications, an increase in natural

enemies’ population, a decrease in the number of pesticide residues in food and a reduction

in farmers’ exposure to the hazardous pesticides (Venilla et al., 2004). Although

insecticides are not applied on Bt cotton against the lepidopteran bollworm complex,

additional applications are necessary to manage sucking insect pests to avoid yield losses

(Xingyuan et al., 2004).

Raghuram and Gupta (2006) conducted field trials with American cotton, cv. Pusa

8–6 to study the bio-efficacy of two neonicotinyl insecticides, acetamiprid at 40 g a.i./ha

22
and imidacloprid at 50 g a.i./ha, against the jassid, Amrasca devastans, reported that

imidacloprid and acetamiprid provided similar levels of protection to cotton crop against

jassid. Saeed et al. (2015) reported that imidacloprid applied at the recommended dose of

5 g/kg seed is effective against A. devastans and appears safer than thiamethoxam for

natural enemies and also enhances plant growth directly.

2.3.1.3. Whiteflies, Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae)

Whiteflies cause damage to cotton plants in two ways. Firstly, by sucking the sap

and secondly by excreting honey dew on which sooty mould grows. Damage from direct

Feeding reduces the photosynthetic activities of the plant and hence the yield. Indirect

damage results from lint contamination with honeydew and associated fungi and through

leaf curl virus disease transmission. The life cycle of whiteflies ranges from 14 to 107 days,

depending on the weather conditions. There are about 12 overlapping generations in a year

and the pest also reproduces parthenogenetically at times. Whiteflies have a vast host range.

The activity of whiteflies ranges from the emergence of seedlings to full-grown crops.

Scanty rainfall and high temperature aggravate the whiteflies’ infestation.

Mustafa (2000) reported that imidacloprid and polo resulted in 72.6% whitefly

mortality. Parrish (2001) and Aslam et al. (2003) reported significant whitefly mortality

with the application of acetamiprid compared to other neonicotinoids. Khattak et al. (2004)

also reported similar results that the imidacloprid and polo significantly reduced the

whitefly population at 24 hrs, 72 hrs and 120 hrs after spray.

Mohammad et al. (2008) reported that imidacloprid gave 87.82% control against

whitefly seven days after treatment. In the experiment conducted by Boricha et al. (2010)

to evaluate the bio-efficacy against whitefly, Bemisia tabaci infesting hybrid cotton

reported that the treatment of acetamiprid (0.005%) was found effective against whitefly

followed by thiamethoxam (0.008%). Saner et al. (2013) reported that the population of

23
whiteflies was effectively suppressed by thiamethoxam 25 WG, acetamiprid 20 SP, fipronil

80 WG followed by fipronil 5 SC, imidacloprid 70 WG, imidacloprid 17.80 SL, lambda-

cyhalothrin 5 SC and triazophos 40 EC.

Afzal et al. (2014) reported that imidacloprid gave the best results (90.87%

mortality after 72 hrs of application), followed by diafenthiuron, acetamiprid,

thiamethoxam and thiacloprid. Kaur et al. (2015) indicated that after three days of the

treatment of triazophos 40 EC (63.22%) followed by the acetamiprid 20% SP (55.61%)

was proved to be most effective. These treatments were significantly superior to the rest of

the treatments, imidacloprid 17.8 SL (53.9%) and thiamethoxam 25 WG (48.7%).

Kar (2017) reported that imidacloprid at 175 ml/ha was the most effective treatment

with 100% control of pest population five days after spray and recorded the minimum

population at 10 and 15 days after spray. This was followed by the treatment of

imidacloprid at 150 ml/ha and thiamethoxam at 200 g/ha. Imidacloprid at 175 ml/ha and

150 ml/ha had a better efficiency against whitefly than thiamethoxam.

2.3.2. Safety of neonicotinoids to natural enemies

While the use of insecticides remains an essential component of integrated pest

management (IPM), biological suppression of insect pests is also considered an equally

important tool. Hence, the protection and preservation of natural enemies of pests are

essential. Among the various entomophagous natural enemies, coccinellids, spiders, green

lacewing and Trichogramma sp. are very important in the cotton ecosystem. The toxicity

of insecticides on parasitoids and predators is reviewed hereunder.

Krishnamoorthy (1985) reported the effect of insecticides belonging to different

groups on eggs, larvae and adults of Chrysoperla scelestes (Banks) and reported that newly

hatched larvae were more susceptible to organophosphate, carbamate and pyrethroid

24
groups of insecticides when compared to egg stage. Larvae and adult stages were highly

susceptible to quinalphos, chlorpyriphos and cypermethrin.

Kapadia and Puri (1991) reported that triazophos, monocrotophos and

fenpropathrin sprayed on cotton had greater residual toxicity to C. carnea larvae than

endosulfan, cypermethrin and fenvalerate. Sharma et al. (1991) reported that cypermethrin

(0.04 kg a.i. per ha), dimethoate (0.40 kg a.i. per ha), monocrotophos (0.40 kg a.i. per ha)

and fenvalerate (0.20 kg a.i. per ha) were toxic to Cheilomenes sexmaculata on chickpea

and lentil.

Rajendran and Gopalan (1996) assessed the effect of endosulfan 35 EC,

monocrotophos 36 WSC and cypermethrin 10 EC at different concentrations on egg

parasitoid T. chilonis and reported the toxicity to the parasitoids even at low concentrations.

JianZhong et al. (1996) reported that imidacloprid displayed good selectivity between

Nilaparvata lugens (Stal) and its natural enemies and it was safe for spider communities.

Toda and Kashio (1997) investigated the toxic effects of 34 insecticides, six

acaricides and nine fungicides on the first instar larvae of C. carnea in the laboratory by

using a direct dipping test and residual contact test. The results reported that all

organophosphate insecticides showed high toxicity, while nitenpyram, imidacloprid and

acetamiprid showed low toxicity in the dipping test, but high toxicity in the residual contact

test.

Schwalbe (1997) reported that the carabid insect population trapped in pitfall traps

placed in sugar beet fields were not affected by imidacloprid treatments. Investigation of

the effect of imidacloprid on the parasitism of Encarsia formosa Gahan on Bemisia

argentifolii Bellows and Perring indicated that parasitoids could develop to the adult stage

on whiteflies infesting imidacloprid treated plants and the parasitism was at low levels

(<10%) due to high levels of whitefly mortality on treated plants.

25
 Imidacloprid at 0.2 kg a.i./ha was found safe for spiders, Cyrtorhinus lividipennis

Reuter and Paederus fuscipes Curtis in the rice ecosystem (Panda and Mishra, 1998). Under

glasshouse conditions, Schoen and Martin (1999) ruled imidacloprid as a harmful

insecticide on the predatory bug, Macrolophus caliginosus (Wagner), in tomatoes. Smith

and Krischik (1999) tested the effect of systemic imidacloprid on Coleomegilla maculata

(DeGeer), a coccinellid predator for pests of interior and landscapes and opined that the use

of imidacloprid may not be suitable for this predator.

Amalin et al. (2000) investigated the selective toxicity of imidacloprid to Hibana

velox (Becker), a predator of citrus leaf miner and reported that imidacloprid applied as

foliar spray had moderate toxicity. In contrast, drench application caused the lowest percent

mortality (10-30%) even at the highest concentration. Patil and Lingappa (2000) reported

the selective toxicity ratio (STR) for tobacco aphid, Myzus nicotianae and its predator,

Cheilomenes sexmaculata (Fab.) and also noted that acephate recorded the lowest STR,

followed by imidacloprid, while endosulfan registered the highest.

James and Vogele (2001) reported that imidacloprid at 0.0053% was highly toxic

to Oechalia schellenbergii Guerin-Meneville and Archimantis latistyla (Serville), partially

toxic to Coccinella transversalis Brown and non-toxic to Pristhesancus plagipennis

Walker and Dicranolaius bellulus Guerin-Meneville. Epperlein and Schmidt (2001)

reported that the seed pelleting of sugarbeet with imidacloprid did not have significant

adverse effects on soil organisms such as spiders, ground beetles and rove beetles.

James and Vogele (2001) reported that imidacloprid at 0.0053% was highly toxic

to Oechalia schellenbergii Guerin-Meneville and Archimantis latistyla (Serville), partially

toxic to Coccinella transversalis Brown and non-toxic to Pristhesancus plagipennis

Walker and Dicranolaius bellulus Guerin-Meneville. Manjunatha and Shivanna (2001)

concluded that imidacloprid was toxic to predatory spiders and mirid bugs in the rice

26
ecosystem, giving a mortality of 30.66% and 77.66% at 100 ml/ha and 400 ml/ha,

respectively.

Pons and Albajes (2001) reported that the densities of syrphids, chrysopids and

Orius sp. in maize fields were very low in plots treated with imidacloprid compared to

untreated control plots. Zenger and Gibb (2001) reported that imidacloprid did not

negatively impact the predatory ant population that feeds on the white grub eggs, the turf

grass pest. Acharya et al. (2002) indicated that at the recommended dose, the newer

molecules, acetamiprid (20 g/a.i.), thiamethoxam (25 g/a.i.), imidacloprid (25 g/a.i.) and

abamectin (20 g/a.i.) were relatively safer to the predatory ladybird beetles.

Mathirajan and Regupathy (2002) reported that imidacloprid at 0.2 ml/l had no

adverse effect on the hatchability of green lacewing, C. carnea eggs. Larval mortality

ranged from 10% to 48.7%, whereas pupation ranged from 51.2% to 73.7%. Compared to

untreated control, adult emergence, longevity and fecundity were lower in treated insects.

Suganthy (2003) reported no significant adverse effect of imidacloprid (17.8% SL)

at 25 g a.i./ha on adult emergence and parasitisation of egg parasitoid, T. chilonis.

Jayaprabhavathi (2005) reported that acetamiprid did not show any ovipositional deterrence

to the above egg parasitoid. Dhaka and Pareek (2007) reported that beneficial insects like

Chelonus blackburni Cameron, Trichogramma achaeae Nagaraja, T. brasiliensis

(Ashmead), Chrysoperla carnea Stephens, Coccinella septempunctata Linnaeus,

Cheilomenes sexmaculata (Fabricius) and spiders were observed to be potential natural

enemies of key cotton pests, playing an important role in the pest management of the cotton

ecosystem.

Natural enemies present in various agro-ecosystems were susceptible to

conventional insecticides. The toxicity of these conventional insecticides has been

demonstrated in laboratory tests and most of them are harmful to different parasites and

27
predators (Michaud and Grant, 2003; Balakrishnan et al., 2009). Acetamiprid was found

to be the least toxic in the residual film method, whereas most toxic in the glass vial method

against Coccinella undecimpunctata (Ahmad et al., 2011).

2.3.3. Phytotoxicity of neonicotinoids

Mote et al. (1993) reported that the imidacloprid 70 WS did not cause any

phytotoxic effect on cotton plants at 5.0, 7.5, 10.0 and 15.0 g per kg seed. Walunj and Mote

(1995) reported that different doses of imidacloprid did not show any phytotoxic symptoms

in tomato plants. Meanwhile, Natwick et al. (1996) reported that the root plug treatments

of imidacloprid at 90, 180, 270 and 360 g a.i./ha and in-furrow treatment at 360 g a.i./ha

exhibited slight phytotoxic symptoms such as leaf tissue necrosis.

Bethke and Redak (1997) noted no phytotoxicity when imidacloprid at 0.002 to 0.02

g a.i./l of water was applied against B. argentifolii on greenhouse vegetables. Swabbing of

imidacloprid at 10, 21, 42 and 88 mg/l of water on chrysanthemum and poinsettia has not

produced any phytotoxic symptoms (Pasian et al., 1997). Foliar application of imidacloprid

17.8 SL at 0.005 and 0.02% did not produce phytotoxic symptoms on cotton (Gupta et al.,

1998).

Imidacloprid 600 FS and 200 SL, even when tested at higher doses, did not cause

any phytotoxicity on the cotton crop (Kumar, 1998); okra (Sivaveerapandian, 2000;

Bhargava and Bhatnagar, 2001); maize (Toit, 2002). But, imidacloprid 600 FS at one mg

a.i./seed produced phytotoxicity in cucumber (Allen et al., 2001) and carrot (Fischer and

Terrettaz, 2002). Premalatha (2001) reported that no phytotoxic symptoms were observed

with thiacloprid 240 SC even at a high dose of 2.0 l/ha.

Kishore et al. (2003) reported that on sorghum and pearl millet, even the 4x doses

of imidacloprid (70 WS and 600 FS formulations) did not exhibit any phytotoxicity

symptoms. Suganthy (2003) noted that even at higher doses, viz., 50 and 100 g a.i./ha,

28
imidacloprid 17.8 SL did not cause phytotoxic symptoms on cotton, bhendi, or chilli except

radish. At the higher dose of 100 g a.i./ha imidacloprid 17.8 SL caused slight phytotoxic

symptoms (Grade 1) in terms of injury to the leaf tip and leaf surface on the radish.

Prasanna et al. (2004) reported that thiamethoxam 70 WS had not affected

germination when applied as seed treatment before sowing. Also, these treatments had not

caused phytotoxic symptoms such as injury to the leaf tip and leaf surface, wilting, vein

clearing, necrosis, epinasty and hyponasty in the NHH-44 cotton variety. Jayaprabhavathi

(2005) reported that acetamiprid 20 SP at a higher dose of 40 and 80 g a.i./ha, than the

normal dose (20 g a.i./ha), did not cause any phytotoxic symptoms in cotton crops. Patil et

al. (2007) reported that two sprays of clothianidin 50% WDG at 20 and 25 g a.i./ha were

safe and no phytotoxicity was noticed on the cotton hybrid DHH-11.

Bhamare and Wadnerkar (2013) reported that application of higher doses of

insecticidal combinations, acetamiprid + cypermethrin, acetamiprid + quinalphos and

acetamiprid + chlorpyriphos resulted only in leaf injury by up to 3 days after spraying,

while other phytotoxic symptoms like wilting, vein clearing, necrosis, epinasty and

hyponasty were not observed on the cotton crop. Bajya et al. (2013) reported no phytotoxic

effect on the cotton crop with thiamethoxam 75% SG applied as soil drench application at

113 and 225 g a.i./ha.

2.3.4. Residues of neonicotinoids

2.3.4.1. Methodology for residue analysis

Analysis of pesticide residues is essential to address rising consumer concerns and

awareness regarding possible contamination problems. A rapid, sensitive and reliable

method using HPLC to determine imidacloprid residues with 75-100% recoveries was

developed by Ishii et al. (1994). The residues from cotton were extracted using 80:20 v/v

acetonitrile: water and analyzed with RP-8 column in HPLC, with the flow rate of 1 ml/min.

29
and a detection wavelength of 265 nm. Kumar (1998) and Sivaveerapandian (2000)

reported the imidacloprid residues on cotton and bhendi, respectively, in HPLC using ODS

2 column.

Frenich et al. (2000) estimated the residues of imidacloprid and its metabolite in

greenhouse air by HPLC with diode-array detection. Pigeon et al. (2000) developed a non-

destructive, rapid and accurate method of estimating imidacloprid on sugar beet seeds using

near-infrared spectroscopy (NIR). Schoning (2001) developed an analytical method for

determining imidacloprid by HPLC with electrospray detection in plant and other materials

such as honey, nectar, bee wax, pollen and leaves of corn, rape, sunflower, trees and horse

chestnuts.

Vidal et al. (2002) described two methods for determining the highly polar

pesticide, acetamiprid in vegetables. Residues were extracted with ethyl acetate and co-

extractives were removed with a graphite carbon-based packing ENVI-carb cartridge.

Analysis was performed with HPLC with post-column photo derivatization, fluorescence

detection, and HPLC-electrospray ionization mass spectrometry. The clean-up step is not

necessary in the last case. The limits of detection were 6μg/l for HPLC – fluorescence

detection using matrix-matched standards and 1.5 μg/l for HPLC-ES-MS detection.

Recovery data obtained by fortifying vegetable matrix at 0.01, 0.1 and 0.5 mg/kg, were

between 65% and 75% for HPLC fluorescence and between 72% and 77% for HPLC

ES-MS.

Acetamiprid was extracted in methanol and derivatized to methyl-6-

chloronicotinate through alkali hydrolysis, potassium permanganate oxidation and then

esterification of diazomethane, followed by column chromatography clean-up and gas

chromatography determination. The limit of detection was 0.01 ppm and the recovery of

fortified samples ranged between 74 and 92% (Tokieda et al., 1997).

30
 Many scientists have dealt with the determination of thiamethoxam residues in

various samples using different types of equipment, viz., LC-MS (Liquid chromatography-

mass spectrometry) (Gonzalez et al., 2008; Pandey et al., 2009); GC (Gas chromatography

by using nitrogen phosphorus detector) (Gabriela et al., 2006); HPLC (High-performance

liquid chromatography) coupled with UV-VIS (Ultraviolet-visible spectroscopy) detector

(Karmakar et al., 2012).

2.3.4.2. Harvest time residues

Gajbhiye et al. (1997) reported the harvest time residues of imidacloprid in

sugarcane juice, leaf and soil for sett dipping in 0.1, 0.15 and 0.2% solutions and furrow

application of imidacloprid 200 SL at one l/ha were found to be below the detectable level

(BDL). Harvest time residues of the imidacloprid as seed treatment insecticide were at BDL

(< 0.1 mg/kg) on black gram and soybean (Gopal et al., 1997). The Association of official

analytical chemists described a method of determining imidacloprid in formulated products

by HPLC with UV detection at 252 nm using propiophenone as an internal standard

(AOAC, 1998).

Residues of imidacloprid (applied both as seed and foliar treatment) on cotton were

detected as BDL in seed, oil and lint (Kumar, 1998). Dikshit et al. (2000) reported that the

residues of imidacloprid in bhendi fruits were found to be 0.08, 0.10, 0.14 and 0.24 mg/kg

from 3, 5.4, 10.8 and 21.6 g a.i./kg seed treatments, respectively, after 55 days of sowing

and became non-detectable after 60 days of sowing. Harvest time residues of imidacloprid

applied as seed and foliar were non-detectable on bhendi (Indumathi et al., 2001; Dikshit

and Lal, 2002; Sivaveerapandian et al., 2002). In seeds, imidacloprid residues were at BDL

except for one sample with 0.01 ppm (Kumar, 1995).

The harvest time residues of quinalphos in lint samples from first picking were at

detectable levels. But, the harvest time residues were at BDL in both lint and seed samples

31
during the third picking (Valarmathi, 1997). The harvest time residues of abamectin 1.9 EC

at 10.8, 14.5, 18.5, 22.5 and 29.0 g a.i./ha were below detectable levels in seed cotton, lint

and oil samples (Sheebajasmine, 2005). According to Dikshit et al. (2005), the harvest time

residues of imidacloprid 70% WG were at BDL in cotton lint and seed samples.

Suganyakanna (2006) reported that acetamiprid 20 SP applied at 10, 20, 40 and 80

g a.i./ha left residues at non-detectable levels in cotton lint, seed and oil at the first, third

and fifth harvests. Mathirajan (2001) reported that thiamethoxam's harvest time residue was

below detectable in rice grains, paddy straw, and seed cotton and lint (Saranya, 2010).

Gupta et al. (2005) studied the persistence of imidacloprid, acetamiprid and thiamethoxam

in gram by seed dressing with imidacloprid at 3 and 6 g a.i./kg seed and foliar spray with

imidacloprid at 20 and 40 g a.i./ha and acetamiprid and thiamethoxam at 25 and 50 g a.i./ha.

The residues in the green plant persisted for 30 days after sowing. But residues were not

detected in green plants after 45 days. Senthilkumar and Regupathy (2008) reported that

the harvest time residues of thiamethoxam were below the detectable limit in the berries

and coffee parchment.

32